Cells doi: 10.3390/cells15212030

Authors: Hajar Dauleh Khalid Hussain

Glucose homeostasis requires coordinated nutrient sensing, membrane excitability, hormone secretion, tissue glucose flux, and mitochondrial metabolism. Ion channels and transporters connect these processes, but mechanistic importance does not necessarily establish human disease causality or clinical actionability. This narrative review integrates human islet electrophysiology with glucose, metal-ion, and mitochondrial transport, distinguishing established causal human disease, replicated genetic susceptibility, limited genetic observations, human functional evidence, and predominantly experimental mechanisms. ATP-sensitive potassium (KATP) channel disorders provide the clearest bidirectional paradigm: activating KCNJ11 or ABCC8 variants can cause neonatal diabetes amenable to sulfonylureas, whereas inactivating variants cause congenital hyperinsulinism. Tissue-selective transporter syndromes, background potassium conductance, intercellular coupling, and organellar calcium handling illustrate additional mechanisms with different levels of human validation. Acquired remodeling in type 2 diabetes may be pathogenic, compensatory, or secondary to metabolic stress. We separate established management from investigational human replacement and preclinical targeting and critically assess functional assays, stem-cell models, transcriptomics, and genome engineering. Priorities are to resolve genotype–phenotype heterogeneity, establish causal direction and disease-stage dependence, and develop tissue-selective interventions with demonstrable patient benefit.

Cells doi: 10.3390/cells15212029

Authors: Geofanny Facicilia Cynthia Retna Sartika Yuni Elsa Hadisaputri Tina Rostinawati

Mesenchymal stem cells (MSCs) are promising platforms for regenerative and immunomodulatory therapies, but their clinical performance is limited by donor heterogeneity, variable potency, and limited persistence. CRISPR-based engineering provides programmable approaches to modify MSC function through genome editing and transcriptional regulation. This review examines CRISPR/Cas9-mediated knockout and knock-in, dCas9-based transcriptional regulation, and emerging base-, prime-, and epigenome-editing technologies for MSC engineering. These approaches can enhance survival and stress resistance, homing and tissue retention, paracrine and extracellular vesicle activity, immunomodulation, immune compatibility, and lineage-associated functions. Key translational challenges include efficient delivery, engineered-population heterogeneity, genomic safety, immunogenicity, phenotypic stability, scalable manufacturing, and mechanism-linked potency assessment. The appropriate engineering strategy should therefore be selected according to the intended therapeutic mechanism and product development requirements. Clinical translation will ultimately depend on demonstrating reproducible functional enhancement, manufacturing consistency, genomic and phenotypic stability, and validated therapeutic potency.

Cells doi: 10.3390/cells15212028

Authors: Marta Peris Juan Sahuquillo Ana Ortuño Verónica Pons Marina Rierola Maria A. Poca

Red blood cell (RBC) transfusion aims to restore tissue oxygen delivery, yet a stored unit’s capacity to unload oxygen depends on 2,3-diphosphoglycerate (2,3-DPG), the principal allosteric modulator of hemoglobin–oxygen affinity. Randomized trials comparing fresher with older units have been consistently null, and all have used storage duration as the exposure variable. We asked whether storage age reflects the metabolic state of individual units. Twenty-four leukodepleted packed RBC units were sampled serially from day 3 to day 42 of standard cold storage. 2,3-DPG was quantified by liquid chromatography–mass spectrometry (LC–MS) with isotopically labelled internal standards, pH was measured in parallel, and trajectories were modelled by linear mixed effects with unit-level random intercepts. At day 3, 2,3-DPG spanned an approximately 18-fold range across units of identical age (0.8–14.7 µmol/g Hb; median 4.0): some were already near-depleted at the earliest sampled timepoint, while others retained near-physiological concentrations. This heterogeneity collapsed rapidly—median values fell to 20% of day-3 levels by day 7 and by more than 95% by day 14 (β = −0.298 µmol/g Hb per day, p < 0.001)—and were below the limit of detection in most units after day 17. pH declined monotonically to 6.40 ± 0.04 by day 42 (β = −0.0163 units per day, p < 0.001). Units of identical storage age therefore differ markedly in the levels of the metabolite that governs oxygen unloading, and are depleted long before the 42-day expiry limit, suggesting that storage duration alone may incompletely capture the metabolic state of stored red blood cell units.

Cells doi: 10.3390/cells15212027

Authors: George Ayoub

Autism spectrum disorder (ASD) research has largely treated early regression, late decline after a period of stability, gradual increases in symptom severity, late diagnosis, and the sex difference in diagnostic timing as separate topics. These phenomena differ in definition, evidence, and plausible mechanism, and diagnostic and recognition processes may explain much of the late-diagnosis and sex-timing findings. This narrative, hypothesis-generating review asks whether a cumulative oxidative and inflammatory burden, arising from environmental chemical exposure, infection and autoimmune activity, and chronic psychosocial stress, and modified by genetic susceptibility, one-carbon metabolism, the gut microbiome, and individual buffering capacity, could contribute to some of these trajectories. We grade the evidence for each link in four tiers (human ASD data, other human data, experimental models, and our hypotheses). Most links rest on associations with ASD risk or on experimental data; no study has linked these exposures to regression or late diagnosis, and oxidative stress may be a consequence or correlate rather than a cause. We therefore define operational terms, derive predictions that could refute the model, and outline the prospective, sex-stratified evidence needed to test it.

Cells doi: 10.3390/cells15212026

Authors: Halle M. Meyers Amira A. Abdellatef Natalia I. Krupenko Baba Mass Venkata Saroja Voruganti Susan Sumner Blake R. Rushing Sergey A. Krupenko

In humans, six common non-synonymous exonic single-nucleotide polymorphisms in ALDH1L1 lead to ten enzyme haplotypes. Since ALDH1L1 is a metabolic regulator and a candidate tumor suppressor, we studied how haplotype variants affect metabotypes, proliferation and motility. Using lentiviral delivery to A549 and PC3 cells, we have generated two cell panels, with each cell line in the panel carrying one of ten ALDH1L1 haplotype variants. Untargeted metabolomics revealed strong differences in metabotypes between cells carrying ALDH1L1 variants. Certain ALDH1L1 variants had decreased ability to regulate one-carbon metabolism and downstream pathways, including amino acid and glutathione metabolism, nucleotide biosynthesis, and the TCA cycle. The direct comparison of A549 and PC3 cells using an untargeted metabolomic dataset also showed that the two cell lines have vastly different metabotypes. In agreement with this finding, the analysis of A549 and PC3 ALDH1L1 haplotype panels suggested that the ALDH1L1-linked metabolic regulation is strongly influenced by specific cellular metabotypes and genetic background. ALDH1L1 variants also exhibited reduced inhibition of cellular proliferation and motility compared to the most common haplotype, which supports further investigation of their potential relevance to individual risk of metabolic disorders.

Cells doi: 10.3390/cells15212025

Authors: Momoko Kawasaki-Takahashi Ryo Tomemori Yoshikazu Matsuoka Tatsuya Kanagawa Yoshiki Yasukochi Koichiro Higasa Hisanori Imai Kaoru Araki-Sasaki Hirofumi Hitomi

Reis–Bücklers corneal dystrophy (RBCD) is an early-onset TGFBI-related corneal dystrophy associated with the TGFBI R124L variant. It is characterized by recurrent corneal epithelial erosion, hyaline-like deposits involving Bowman’s layer and the anterior stroma, and progressive subepithelial corneal opacity. However, how mutant TGFBI protein (TGFBIp) accumulates in the cornea remains unclear. In this study, we established induced pluripotent stem cells (iPSCs) from two affected members of an RBCD family and one unaffected family member. The patient-derived lines retained the R124L variant, whereas the line established from the unaffected mother did not. All three lines showed pluripotency and trilineage differentiation, had normal karyotypes, and were confirmed by STR analysis to match their donors. After 4 weeks of SEAM induction, all three iPSC lines developed early SEAM-like cultures containing PAX6- and p63-positive cells, demonstrating their capacity for early differentiation toward the corneal epithelial lineage. TGFBIp was also detected in extracellular and pericellular regions by immunofluorescence. In silico analysis detected no major structural difference between WT and R124L TGFBIp in the high-confidence CROPT and FAS1 domain regions, whereas surface analysis showed local changes in electrostatic potential and molecular lipophilicity around residue 124. These lines can be used to study TGFBIp deposition and epithelial–stromal interactions in RBCD.

Cells doi: 10.3390/cells15212024

Authors: Renfang S. Taylor

The (pro)renin receptor ((P)RR), encoded by ATP6AP2, participates in kidney development and renal physiology through several potentially distinct activities: ligand-associated signaling, V-ATPase-related cellular functions, and generation of soluble (P)RR (s(P)RR). Experimental lineage-targeted ATP6AP2 perturbation supports requirements in ureteric bud branching and stromal–epithelial interactions, and a ureteric bud deletion model implicates Dot1/H3K79-associated changes. Nevertheless, deletion phenotypes do not by themselves establish that renin/prorenin binding causes the developmental defects. Wnt/V-ATPase interactions provide a mechanistic framework, whereas proposed links to GDNF/RET, Notch, PI3K/AKT, and metabolic pathways vary in evidentiary support and should not be interpreted as proven kidney-specific regulatory axes. Evidence for s(P)RR in sodium handling and adult renal injury must likewise be distinguished from its uncertain role in nephrogenesis. This review evaluates the strength and limitations of experimental findings, compares available developmental models, and outlines approaches to distinguish ligand-dependent actions from ATP6AP2/V-ATPase-associated functions. An association between abnormal nephrogenesis and later kidney disease is biologically plausible, but direct longitudinal evidence following developmental ATP6AP2 disruption remains limited.

Cells doi: 10.3390/cells15212023

Authors: Kamila Adamczuk Grzegorz Adamczuk Michał Kiełbus Matthias Nees Andrzej Stepulak

Head and neck squamous cell carcinoma (HNSCC) develops resistance to cisplatin. We investigated whether JIB-04, an inhibitor of Jumonji histone demethylases, enhances cisplatin activity in established HNSCC cell lines. Four UT-SCC cell lines originating from primary tumors and matched lymph-node metastases were treated with JIB-04 and cisplatin, alone or combined. Viability was assessed by MTT assay, and drug interactions were evaluated using the Highest Single Agent model. In UT-SCC-24A cells, we assessed BrdU incorporation, LDH release, cell-cycle distribution, and apoptosis. RNA sequencing was performed after 24 h at the same drug concentrations used in functional assays. JIB-04 reduced viability in all HNSCC lines at submicromolar concentrations, with moderate effects in BJ fibroblasts. HSA analysis identified a localized positive interaction in UT-SCC-24A, where the overall HSA score was 4.25, whereas local scores reached approximately 16–18 within a narrow low-dose range. Co-treatment further reduced DNA synthesis and increased S/G2-M accumulation and apoptosis, without significantly increasing LDH release. RNA sequencing revealed JIB-04-associated induction of integrated stress response/ATF4- and DNA damage response-related genes, alongside suppression of proliferation- and cell-cycle-associated genes. The combination retained these transcriptional features. These findings support the further investigation of JIB-04–cisplatin combinations in selected HNSCC cell-line models.

Cells doi: 10.3390/cells15212021

Authors: Yuanjun Xin Jie Chen Hongxia Zhang Robert P. Ostrowski Yidan Liang Jun Zhao Xiang Xiang Fuming Liang Wenqiao Fu Hao Huang Xintong Wu Jun Su Jiewen Deng Zhaohui He

The journal retracts the article titled “Dexras1 Induces Dysdifferentiation of Oligodendrocytes and Myelin Injury by Inhibiting the cAMP-CREB Pathway after Subarachnoid Hemorrhage” [...]

Cells doi: 10.3390/cells15212022

Authors: Liyuan Fu Rui Guo Qingyu Zhang Qichun Zhang

Metabolic dysfunction-associated steatohepatitis (MASH) involves hepatocellular injury, myeloid activation, and fibrotic remodeling that may resolve unevenly following metabolic improvement. Whether previous injury leaves a functionally consequential innate immune imprint remains unresolved. This review evaluates trained immunity as a candidate mechanism linking exposure history to persistent hepatic myeloid responsiveness. We distinguish continued activation by lipotoxic and stromal signals from responses retained after stimulus withdrawal, macrophage replacement, and hematopoietic progenitor programming. Evidence from Kupffer cells, monocyte-derived macrophages, and lipid- or scar-associated populations reveals divergent inflammatory, efferocytosis, and matrix remodeling functions. Receptor-dependent lipid handling intersects with glycolysis, citrate-derived acetyl-CoA, serine and one-carbon metabolism, NAD+ availability and histone methylation, acetylation, and lactylation. These pathways explain how metabolic inputs regulate inflammatory competence, but their alteration during active MASH does not establish immune memory. Rested monocyte training models provide stronger temporal evidence than most disease-specific studies, whereas protective TREM2-dependent responses caution against indiscriminate suppression of disease-associated macrophages. Establishing a contribution to MASH recurrence requires lineage-resolved investigations spanning injury, regression, and rechallenge, with interventions separating memory establishment, maintenance, and effector output. If exposure-dependent myeloid memory contributes causally to MASH recurrence, targeting maladaptive responsiveness could complement injury control while preserving hepatic repair.

Cells doi: 10.3390/cells15212020

Authors: Jingwei Kang Shigang Ding Xingting Guo

Gastric precancer is conventionally staged as a linear histological sequence, but it develops within a patchy, continuously remodeling mucosa. This narrative review reframes gastric carcinogenesis as a multiscale spatial process in which epithelial states, clonal gland dynamics, immune–stromal neighborhoods, regional Helicobacter pylori (H. pylori) and non-H. pylori colonization, and metabolic or physicochemical gradients may jointly shape local risk. Gland-resolved studies show that metaplasia may expand through fission and remain clonally continuous with dysplasia in selected lesions, whereas broad genomic and epigenetic field alterations may also support independent tumor emergence. Single-cell and spatial analyses identify hybrid, stem-like, and proliferative epithelial states embedded within evolving multicellular niches, although most evidence remains cross-sectional and biased toward established cancer. We therefore propose a non-deterministic model in which injury drives gland loss and niche remodeling; metaplastic clones expand and diversify; local ecological pressures may select high-risk states; and only a minority of glands cross the neoplastic threshold. This remains a working hypothesis, particularly for niche-driven selection of human precancerous clones. Clinically, the framework is consistent with the prognostic value of metaplastic extent, corpus involvement, and the Operative Link on Gastritis Assessment (OLGA) and Operative Link on Gastric Intestinal Metaplasia Assessment (OLGIM) systems, while defining the longitudinal, topographically annotated, gland-resolved studies needed to determine whether spatial risk maps add clinical value beyond established staging.

Cells doi: 10.3390/cells15212019

Authors: Valeriia S. Shtol Sophia P. Konovalova Kirill A. Arsentiev Yulia I. Taranukha Pavel E. Musienko

Biomaterials for neural tissue engineering, cell delivery, and chronic neural interfaces are conventionally characterized through physicochemical descriptors such as stiffness, topography, surface chemistry, and degradation rate. However, their biological effects are mediated by the material–tissue interface, which evolves through protein adsorption, extracellular matrix ligand presentation, and progressive cellular remodeling and determines how these design parameters are translated into neuronal and glial responses. This review synthesizes current evidence on how neurons, astrocytes, and microglia sense the material–tissue interface, how integrin-dependent adhesion complexes, mechanosensitive Piezo1 channels, and downstream FAK/Src, RhoA/ROCK, and YAP/TAZ pathways transduce interfacial cues into intracellular signaling, and how key design variables (mechanical properties, architecture, surface chemistry, electrical conductivity, and degradation) can be tuned to direct these processes in regenerative scaffolds and chronic neural interfaces. Particular attention is given to studies distinguishing direct neuronal effects from glia-mediated modulation, since concurrent changes in neuronal and glial markers are often reported without establishing causal neuron–glia interaction. We further discuss methodological limitations that constrain current interpretations of biomaterial-induced neuron–glia crosstalk, including discrepancies between 2D, 3D, and in vivo models and insufficient durations of chronic observation. We argue that functional biomaterial design for neural applications should move from optimizing isolated cellular readouts toward multicellular experimental frameworks capable of resolving neuron–glia–ECM interactions over physiologically relevant timescales.

Cells doi: 10.3390/cells15212017

Authors: Daria D. Vlasova Anastasia A. Kotikova Viacheslav A. Shmarov Tatiana A. Shigueva Ilya V. Rukavishnikov Elena S. Tomilovskaya Milos Ljubisavljevic Dhifaf Sarhan Nik V. Kuznetsov

Background: The health of astronauts and success of space missions are contingent on resilience to the cumulative stressors of the space environment, including radiation, isolation, confinement and microgravity. While these factors are known to disrupt physiological function at molecular and cellular levels, the isolated impact of microgravity itself remains poorly defined. Objective: To explore in vivo effects of long-term Simulated MicroGravity (SMG) on T cell transcriptomes by analyzing gene expression changes in peripheral blood T cells of healthy volunteers. Methods: A three-week SMG exposure was completed by ten healthy male participants, during which peripheral blood specimens were harvested at five critical junctures (covering baseline, intra-exposure and recovery stages). Post-collection, T lymphocytes were enriched, and their total RNA was extracted. Transcriptome analysis was performed by RNA-seq, the gold standard for transcriptomics owing to its unbiased approach to gene detection and quantification, which provides genome-wide quantification with global normalization and, through a genome-wide analytical approach, the statistical framework to detect differentially expressed genes with controlled error rates. Following next-generation sequencing, a rigorous bioinformatics framework was employed to analyze and interpret the volunteers’ global gene expression dynamics. Results: Ground-based SMG induces widespread deregulation of gene-regulatory circuitry within T lymphocytes. SMG is associated with transcriptome shifts in several key molecular pathways in vivo including those involved in cell proliferation, cell cycle regulation, maintenance of genome integrity, as well as oncogenic and tumor suppressor gene networks. Corresponding hub genes were differentially expressed in the majority of tested healthy volunteers. Conclusions: SMG perturbs essential intracellular gene-regulatory networks in human T cells in vivo. The mechanical unloading imposed by microgravity, even in the absence of detectable systemic effects, was sufficient to deregulate core gene networks, upset the transcript-level equilibrium of central molecular pathways and alter associated gene expression profiles.

Cells doi: 10.3390/cells15212018

Authors: Wojciech Homenda Dominika Strzała Aleksandra Połom Rafał Moraczyński Krzysztof Homenda Sylwia Paszek Natalia Potocka-Wojtowicz Izabela Zawlik

Background/Objectives: Immune thrombocytopenia (ITP) involves immune-mediated platelet destruction and impaired platelet production. Long non-coding RNAs (lncRNAs) regulate immune responses, but their clinical relevance in ITP remains uncertain. This narrative review examines disease-associated expression and mechanistic findings, distinguishing translational potential from established clinical utility. Methods: English-language literature identified through PubMed, Scopus, and Google Scholar was synthesized narratively. Eight named lncRNAs were prioritized because the cited ITP studies provided transcript-specific patient data or functional evidence; exploratory expression profiling and evidence from other diseases were considered separately. Study design, sample size, specimen type, treatment exposure, and validation status informed interpretation. Results: The selected transcripts were HCP5, NEAT1, lnc-DC, IFNG-AS1, MEG3, GAS5, MALAT1, and THRIL. Among the clinical studies with reported sample sizes, ITP groups generally included 20–90 patients, except for a GAS5 polymorphism study involving 302 patients. An independent prognostic association for low HCP5 expression was reported in a single study of 40 patients and has not been externally validated. Other studies described exploratory associations with bleeding, disease duration, or treatment response. Distinguishing primary from secondary ITP and selecting patients for splenectomy are not established applications. Cell experiments and a mouse model support mechanistic hypotheses but do not demonstrate therapeutic efficacy in humans. Conclusions: The evidence supports biological plausibility and translational investigation, rather than routine diagnostic, prognostic, or treatment-guiding use. Prospective validation in clinically representative cohorts and standardized assays are required.

Cells doi: 10.3390/cells15212016

Authors: Katharina Schwarz Carlotta Rickert Fabian L. Kendlbacher Oliwia Miłek Christian Behm Christina Schäffer Oleh Andrukhov

Periodontal health and disease are shaped by a dynamic interaction between the subgingival microbiome and the host immune system, in which gingival macrophages contribute to tissue homeostasis, disease progression, and inflammation resolution. Here, we investigated how three periodontitis-associated bacteria, Fusobacterium nucleatum, Porphyromonas gingivalis, and Tannerella forsythia, shape the polarization of human macrophages in vitro. THP-1 monocytes were differentiated into unpolarized M0 macrophages, or polarized toward the M1 or M2 phenotype, and infected with each bacterium at different multiplicities of infection. Polarization was assessed from the surface expression of CD80 (M1) and CD209 (M2) and from pro- and anti-inflammatory cytokine production, measured by a multiplex assay. The response to P. gingivalis differed strikingly from the two other species. Porphyromonas gingivalis suppressed both M1 and M2 polarization and reduced the levels of several pro- and anti-inflammatory cytokines. In contrast, F. nucleatum and T. forsythia enhanced CD80 expression and pro-inflammatory cytokine production in M1 macrophages and modestly increased pro-inflammatory mediators in M2 macrophages. These findings point to a distinct role for P. gingivalis as a keystone pathogen that subverts the innate immune response, and for F. nucleatum and T. forsythia as periodontal pathobionts.

Cells doi: 10.3390/cells15212015

Authors: Andrea Matucci Michael Diomaiuti Amelia Morrone Pasquale Esposito Federico Pieruzzi Maurizio Pieroni Emanuele Vivarelli Elena Biagini Raffaello Ditaranto Maria Alessandra Schiavo Calogero Lino Cirami Leonardo Caroti Egrina Dervishi Ilaria Romani Cristina Sarti Mario Mangeri Barbara Trezzi Letizia Roggero Claudia Sgattoni Alessia Gennarini Giancarlo la Marca Giulia Forni Giacomo Mascia Marina Rinaldi Annamaria Del Franco Alessia Argirò Sandro Feriozzi Matteo Accinno Alessandra Vultaggio

(1) Background: Anti-drug antibodies (ADAs) may develop during enzyme replacement therapy (ERT) for Fabry disease and potentially affect biochemical response. Evidence have been derived mainly from males, while comparisons between agalsidase-alpha and agalsidase-beta remain limited by differences in ADA assays. The aim of the study is to evaluate ADA prevalence and titers in patients with Fabry disease treated with agalsidase-alpha or agalsidase-beta and to investigate associations with sex, GLA genotype, residual α-galactosidase A activity, and lyso-Gb3 levels. (2) Methods: This multicenter observational study included 117 patients with Fabry disease receiving ERT (50 males, 67 females), treated with agalsidase-alpha (n = 52) or agalsidase-beta (n = 65). ADAs were measured using an in-house enzyme-linked immunosorbent assay, and lyso-Gb3 by liquid chromatography–tandem mass spectrometry. GLA variants were centrally classified as predicted null-type or predicted non-null-type on the basis of the variant class, and α-galactosidase A activity at diagnosis was normalized to the lower limit of the assay reference interval. Multivariable regression analysis identified factors associated with ADA positivity and lyso-Gb3 levels. (3) Results: ADAs were detected in 55/117 patients. Positivity was more frequent in males than females, with no statistically significant difference between agalsidase-alpha and agalsidase-beta. Predicted null-type variants were associated with higher ADA positivity and titers, particularly in males. In multivariable models, male sex and predicted null-type variants independently predicted ADA positivity, whereas higher normalized α-galactosidase A activity was associated with lower odds. ADA positivity was not independently associated with lyso-Gb3. (4) Conclusions: ADA detection was primarily associated with male sex and predicted null-type GLA variants; no statistically significant association with the agalsidase administered was found. The high prevalence of binding ADAs in females warrants confirmation using orthogonal and functional assays.

Cells doi: 10.3390/cells15212014

Authors: Xinxin Quan Xue Xue Huilan Ma Lei Yang Chen Chen Yu Liu Kejie Yao Hui Yang Rongxiang Wang Liya Shi Lun Suo Qiuju Chen Lihua Sun

In the published publication [...]

Cells doi: 10.3390/cells15212013

Authors: Jenny Pakhrin Rana Joseph L. Graves Radiah C. Minor Jenora T. Waterman

In livestock production settings, repetitive exposure to organic dust (OD) is a risk factor for chronic bronchitis (CB). Macrophages and lymphocytes cooperate to defend the lungs against inhaled dust, but when chronically activated, their actions contribute to chronic inflammation, oxidative stress, and tissue remodeling. Despite the strong body of literature examining the effects of OD exposure on leukocytes and lungs, the contribution of macrophages and lymphocytes to organic dust-associated chronic bronchitis is not fully understood. This review examines the role of macrophages and lymphocytes in organic dust-associated chronic bronchitis. To identify the body of literature addressing this goal, we conducted Boolean searches with keywords (e.g., bronchitis, leukocytes, organic dust) across four major databases: PubMed, Science Direct, Scopus, and Web of Science. From a total of 25,488 hits from all time, 18 research articles (from 2015 to 2025) provided contextual connection for explaining organic dust-associated chronic bronchitis. The results showed continuous or repetitive exposure to OD or its components (e.g., lipopolysaccharide) causes prolonged activation of macrophages and lymphocytes, which leads to overexpression of pro- and anti-inflammatory cytokines and chemokines via Toll-like receptor (TLR)4, TLR2, nuclear factor kappa B, protein kinase C, Extracellular Signal-Regulated Kinase (ERK), and Mitogen-Activated Protein Kinase (MAPK) pathways. Unresolved lung inflammation drives pathologic changes including mucus hypersecretion, airway remodeling, and onset of chronic bronchitis symptoms. Addition of micronutrients or other anti-inflammatory treatment helps to mitigate OD-induced lung inflammation. Defining the cellular and molecular contributions to organic dust-associated chronic bronchitis is essential for the development of targeted therapies that prevent or relieve prolonged airway inflammation.

Cells doi: 10.3390/cells15212012

Authors: Jia-Xi Zhang Yi-Kai Zou Xian-Guang Lin Shao-Hua Qi Jia-Wei Min

Ca2+/calmodulin-dependent protein kinase kinase 2 (CaMKK2) couples calcium signaling to metabolic, inflammatory, and transcriptional pathways and has been implicated in a broad range of central nervous system (CNS) disorders. However, its effects vary across disease settings, and the mechanisms underlying this context dependence remain poorly defined. This narrative review examines genetic, transcriptomic, biochemical, cellular, and in vivo evidence on the roles of CaMKK2 across CNS disorders, with emphasis on signaling pathways, cell type-specific functions, and implications for therapeutic targeting. CaMKK2-associated pathways influence neurovascular responses, synaptic function, protein homeostasis, inflammation, and tumor progression through signaling networks involving AMPK, CaMK1, CaMK4, and Akt. CaMKK2 can exert distinct, and in some settings opposing, effects depending on cell type, downstream pathway engagement, intervention timing, and pathological context. Pharmacological studies remain limited by incomplete selectivity, insufficient characterization of CNS pharmacokinetics, and context-dependent biological effects. Moreover, much of the mechanistic evidence derives from experimental models, and validated biomarkers for clinical translation remain limited. Overall, CaMKK2 emerges as a context-dependent signaling node linking calcium signaling to metabolic and inflammatory pathways in CNS disorders, with therapeutic potential but substantial challenges for therapeutic translation.

Cells doi: 10.3390/cells15212011

Authors: Ibrahim Antoun Georgia R. Layton Riyaz Somani Giovanni Mariscalco G. André Ng Mustafa Zakkar

Chronic coronary syndromes (CCSs) are no longer defined only by fixed stenosis of the epicardial coronary arteries. CCSs reflect a dynamic biological process. Key contributors include endothelial activation, lipid retention, macrophage foam-cell formation, vascular smooth muscle cell phenotypic switching, and extracellular matrix remodelling. Microvascular rarefaction, perivascular adipose tissue inflammation, cardiometabolic comorbidity, and biological ageing add further complexity. This broader cellular and molecular model provides a rationale for artificial intelligence (AI), because conventional tools often assess anatomy, physiology, inflammation, and systemic biology in isolation. In this narrative review, we examine how AI and machine learning may support a more mechanistic and individualised approach to CCS by linking cellular pathways to measurable clinical signatures. We focus on AI-enabled plaque phenotyping, non-invasive physiology and ischaemia modelling, assessment of coronary microvascular dysfunction, perivascular inflammation mapping, and the integration of multi-omics with cardiometabolic risk profiling. Most of these tools, however, provide imaging-based or data-derived surrogates rather than direct measurements of endothelial activation, inflammation, or calcification biology. Within this limitation, current evidence suggests that AI can quantify subvisual high-risk plaque features, support non-invasive flow assessment, detect imaging patterns associated with microvascular dysfunction and perivascular inflammation, and, in research settings, relate molecular signatures to long-term cardiovascular risk. Clinical adoption remains constrained by retrospective study designs, limited external validation, heterogeneous imaging protocols, gaps in explainability, regulatory uncertainty, and algorithmic bias across sex, ethnicity, age, and resource settings. Future work should prioritise prospective trials, transparent reporting, federated validation, clinically interpretable outputs, and integration with guideline-based decision pathways. The greatest value of AI in CCS is likely to lie in augmenting mechanistic risk stratification and enabling precision prevention for patients whose risk is poorly captured by anatomy alone.

Cells doi: 10.3390/cells15212010

Authors: Doste R. Mamand

Extracellular vesicles (EVs) are promising nanocarriers for therapeutic delivery; however, the factors influencing their interactions with recipient cells remain incompletely understood. This study investigated recipient-cell-specific EV association using mNG-CD81-labelled EVs derived from HEK293T, SKBR-3, and Yummer cells. EV-associated fluorescence was assessed by flow cytometry following incubation with human epithelial, immune, and murine melanoma cell lines at different doses and time points. Marked differences were observed across recipient-cell types. Liver-derived Huh7 and HepG2 cells exhibited the highest percentages of fluorescence-positive cells among human epithelial cells, whereas parental HEK293T cells showed the lowest. THP-1 cells exhibited greater EV-associated fluorescence than Jurkat cells, while Yummer melanoma cells showed higher fluorescence than B16F10 cells following exposure to HEK293T-derived EVs. Similar recipient-cell patterns were observed with SKBR-3- and Yummer-derived EVs. Notably, Yummer cells exhibited relatively high association with their own EVs, demonstrating that low parental-cell association is not universal. EV-associated fluorescence generally increased with dose and incubation time. Although these findings highlight recipient-cell specificity, the relative contributions of EV donor origin and recipient-cell characteristics remain unresolved. Moreover, flow cytometry cannot distinguish surface-associated from internalized EVs. Together, these findings identify recipient-cell specificity as an important determinant of EV-associated fluorescence, providing a foundation for understanding EV–cell interactions and informing the development of targeted EV-based delivery strategies.

Cells doi: 10.3390/cells15212009

Authors: Xi Deng Haruka Kato Eri Matsuo Eiji Kawamoto Motomu Shimaoka Eun Jeong Park

Sepsis is a life-threatening syndrome characterized by dysregulated systemic inflammation and multiple organ dysfunction, yet the molecular mechanisms underlying its progression remain incompletely understood. Our previous studies demonstrated that microRNA-511-3p (miR-511-3p) is upregulated in both an experimental sepsis model and patients with sepsis, suggesting its potential role in the pathogenesis of sepsis-associated inflammation. Because patched 1 (PTCH1), a validated target of miR-511-3p, serves as a key negative regulator of hedgehog (HH) signaling, we hypothesized that miR-511-3p promotes inflammatory responses during sepsis by suppressing PTCH1 expression and consequently activating HH signaling. To investigate this mechanism, we examined multiple immune cell populations, including small intestinal epithelial cells (S-IECs), small intestinal lamina propria macrophages (SI-LPMs), and liver macrophages (Liv-Ms), in a lipopolysaccharide (LPS)-induced murine sepsis model, as these cell populations may contribute to both local intestinal and systemic inflammatory responses during sepsis. The systemic administration of a miR-511-3p inhibitor (511-Inh) significantly reduced the expression of the proinflammatory cytokines interleukin 6 (IL-6) and tumor necrosis factor (TNF), as well as Hedgehog-interacting protein (HHIP), a canonical HH target gene. The inhibition of miR-511-3p restored PTCH1 expression in the intestines, liver, and lung and increased its mRNA levels in S-IECs, SI-LPMs, and Liv-Ms. These changes were accompanied by a reduced expression of HH pathway-related genes. Furthermore, miR-511-3p inhibition attenuated sepsis-induced liver injury, as evidenced by improved histopathological findings and decreased plasma levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT). To further determine the contribution of HH signaling to sepsis-associated inflammation, septic mice were treated with vismodegib, a selective inhibitor of smoothened (SMO). The pharmacological inhibition of SMO considerably reduced the expression of Smo, Gli, Hhip, Il6, and Tnf in the small intestine, large intestine, liver, and lung, confirming that aberrant HH pathway activation amplifies inflammatory responses during sepsis. Collectively, these findings suggest that miR-511-3p drives sepsis-associated inflammation, at least in part, by suppressing PTCH1 expression and thereby promoting HH signaling. Conversely, inhibiting miR-511-3p restores PTCH1 expression, dampens HH pathway activity, and mitigates systemic inflammation and sepsis-induced organ injury. Thus, the miR-511-3p/PTCH1/HH signaling axis may represent a promising therapeutic target for sepsis.

Cells doi: 10.3390/cells15212008

Authors: Simao Xu Jingzhi Zhang Benxiang He

Background: Tendon stem cells (TSCs) contribute to tendon maintenance and repair through their self-renewal capacity and multilineage differentiation potential. Platelet-rich plasma-derived extracellular vesicles (PRP-EVs) may regulate tissue repair; however, their effects on TSC cellular activity and lineage-associated marker expression remain unclear. Objective: To investigate the effects of different concentrations of PRP-EVs on CCK-8 signal and the expression of tendon-associated and alternative lineage-associated markers in TSCs. Methods: Rat TSCs were isolated from Achilles tendon tissue by enzymatic digestion and characterized using flow cytometry, immunofluorescence staining, colony formation assays, and multilineage differentiation assays. PRP-EVs were isolated from pooled rat PRP and characterized using transmission electron microscopy (TEM), dynamic light scattering (DLS), and Western blotting. TSCs were treated with PRP-EVs at concentrations of 0, 5, 20, and 40 μg/mL. CCK-8 absorbance was measured as an indicator of cellular metabolic activity. The mRNA and protein expression levels of tendon-associated markers (ScxA, Col I, and Tnmd) and alternative lineage-associated markers (Osx, RUNX2, SOX9, and PPARγ) were assessed using quantitative real-time PCR and Western blotting, respectively. Results: TEM revealed vesicle-like structures in the PRP-EV preparation, with a mean particle diameter of 198.6 ± 94.5 nm measured by DLS. Western blotting detected the EV-associated proteins TSG101, CD9, and CD81. PRP-EV treatment increased the CCK-8 signal, with 40 μg/mL generally producing the strongest overall response among the concentrations tested, although differences between concentrations were not consistently significant. Treatment was also associated with increased expression of tendon-associated markers and generally decreased expression of alternative lineage-associated markers. Conclusion: PRP-EV treatment increased the CCK-8 signal and altered the expression of tendon-associated and alternative lineage-associated markers in cultured rat TSCs. These findings indicate that PRP-EVs modulate the molecular phenotype of TSCs in vitro.

Cells doi: 10.3390/cells15212007

Authors: Ching-Thong Choo Zhen-Kai Ngian Chin-Tong Ong

Active H3K27ac marks are dynamically remodelled during brain aging, but their cell-type-specific regulatory roles remain unclear. We integrated genome-wide H3K27ac profiles and transcriptomes from aging Drosophila heads, mouse NeuN+ forebrain neurons, and human NeuN+/NeuN− frontal cortex cells. In Drosophila, age-gained differential (Δ) H3K4me1 and H3K27ac peaks were associated with RNA processing and immune response genes, respectively, suggesting distinct regulatory programs. Despite limited biological replicates, mouse analyses revealed a strong association between promoter-associated ΔH3K27ac and synaptic gene expression in young neurons. Age-related gene-body H3K27ac loss coincided with increased expression of genes involved in GTPase signalling and vesicular transport. In humans (n = 29), NeuN− cells exhibited more age-associated ΔH3K27ac alterations than NeuN+ cells. Genes associated with age-gained peaks were enriched for gliogenesis and amyloid precursor protein metabolic processes, while age-related transcriptional changes associated positively with super-enhancer activity. Comparison with pan-tissue aging signatures revealed shared and cell-type-specific age-related ΔH3K27ac patterns in NeuN+ and NeuN− cells. Integrative analyses showed that age-gained ΔH3K27ac peaks in NeuN− cells overlapped with AD-associated ΔH3K27ac peaks, differentially expressed AD genes, and AD GWAS-linked variants, including those mapped to BIN1 and CR1. These findings highlight extensive H3K27ac remodelling in aging NeuN− cells and its convergence with AD-associated molecular alterations.

Cells doi: 10.3390/cells15212006

Authors: Alka Ashok Singh

Glioblastoma (GBM) remains one of the most aggressive and lethal brain tumors, with limited treatment success despite standard multimodal therapy. Oxidative stress and dysregulated brain-derived neurotrophic factor (BDNF) signaling contribute to GBM progression, therapeutic resistance, and treatment-related neurotoxicity. Therefore, this review aims to examine the complex interplay among antioxidants, BDNF/TrkB signaling, and redox imbalance in GBM, focusing on the potential biological activities of natural antioxidants, particularly polyphenols such as curcumin, resveratrol, and epigallocatechin-3-gallate (EGCG). Preclinical studies suggest that these compounds may modulate oxidative stress, influence glioma cell survival, and alter signaling pathways associated with chemoradiotherapy sensitivity and neuronal protection. Nevertheless, direct evidence that BDNF/TrkB signaling mediates the antitumor effects of these antioxidant compounds in glioblastoma remains insufficient, and observations from non-GBM models should be regarded as mechanistic hypotheses rather than validated therapeutic mechanisms. However, clinical evidence supporting their efficacy in patients with GBM remains limited and inconclusive. Human findings from observational cohorts and clinical trials involving non-GBM populations provide indirect insights into antioxidant-related cognitive and redox outcomes but cannot establish therapeutic benefits in glioblastoma. Furthermore, the dual role of BDNF/TrkB signaling in neuronal protection and tumor progression raises concerns regarding generalized BDNF enhancement. Challenges involving bioavailability, optimal dosing, tumor-specific effects, and potential interactions with conventional treatments further limit clinical translation. Future research should prioritize GBM-specific randomized controlled trials, biomarker-guided strategies, and the simultaneous evaluation of neuroprotective and oncological outcomes. Targeting the antioxidant-BDNF-redox axis remains an experimental therapeutic approach that requires rigorous validation before clinical implementation.

Cells doi: 10.3390/cells15212005

Authors: Tsugumi Tanaka Hiroshi Nango Komugi Tsuruta Hiroko Miyagishi Tadashi Saigusa Kosuke Sugawa Yasuhiro Kosuge

Oxidative stress and proteotoxic endoplasmic reticulum (ER) stress are interconnected cellular stress processes implicated in neuronal injury. J147 reportedly exerts neuroprotective effects through ATP synthase α-subunit (ATP5A)-associated mitochondrial mechanisms, but whether it protects cells from death under proteotoxic ER stress through the same mechanism remains unclear. Here, we directly compared curcumin, CNB-001, and J147 under glutamate-induced oxytosis and tunicamycin (TM)-induced ER stress in murine hippocampal HT22 cells. MTT reduction assays showed that all three compounds attenuated glutamate-induced cell death, whereas only J147 protected cells from TM-induced cytotoxicity at a submicromolar range. LIVE/DEAD staining confirmed the protective effects of J147 against both stressors. At the molecular level, J147 suppressed the TM-induced upregulation of the pro-apoptotic transcription factor C/EBP homologous protein (CHOP) at the mRNA and protein levels without suppressing the adaptive ER chaperone glucose-regulated protein 78. ATP5A knockdown attenuated glutamate-induced oxytosis but did not protect against TM-induced cell death, and J147 retained its protective effect after ATP5A knockdown. These findings support stressor-dependent protective actions of J147 involving an ATP5A-associated mechanism against oxidative stress and an ATP5A-independent mechanism associated with suppression of CHOP induction. J147 may therefore provide a useful pharmacological tool for investigating neuronal responses to interconnected oxidative and proteotoxic stress.

Cells doi: 10.3390/cells15212004

Authors: Patrick J. Stiff Cheryl M. Czerlanis Ronald K. Potkul William Small Maureen L. Drakes

Ovarian cancer is a lethal gynecologic malignancy with limited effective treatment options. It is expected that in 2026 there will be about 21,010 new cases of ovarian cancer diagnosed and about 12,450 deaths within the United States. Unfortunately, conventional therapies such as chemotherapy and radiation have limited efficacy for high-grade serous ovarian cancer (HGSOC) and also produce significant toxicity. HGSOC is characterized by an ovarian tumor microenvironment (TME) consisting of immune inhibitory T regulatory cells and inhibitory tumor-associated macrophages, including myeloid-derived suppressor cells (MDSCs) and M2 cells, which confer immune suppression. Immunocompetent T cells, tumor-suppressing M1 cells, natural killer cells, and other cellular subsets are also present. The TME is further shaped by soluble molecules, extracellular matrix, and other structural components, as well as the presence of blood vessels and ascitic fluid in the peritoneal cavity, which aid in the distribution of cancer cells to the omentum where more lesions soon develop. This review outlines the heterogenous immune cell subsets in the ovarian TME and other structural targets that promote or reduce cancer burden. Most single-agent immune strategies have shown limited efficacy in unselected HGSOC populations due to the complex immune-suppressive nature of the TME. Here, we also discuss novel combination treatments in clinical trials or in clinical practice for HGSOC. Advances in understanding the ovarian TME will reveal new immunotherapy and drug targets for translational medicine and aid in determining effective drug and immunotherapy combinations, which may ultimately lead to improved overall survival in HGSOC.

Cells doi: 10.3390/cells15212003

Authors: Arun Pal Banaja P. Dash Marc Uhlarz Jonas Gronemann Julia Japtok Hannes Glaß Jens Pietzsch Joachim Wosnitza Katrin Sameith Andreas Dahl Andreas Hermann Richard H. W. Funk Thomas Herrmannsdörfer

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease hallmarked by progressive loss of neuromuscular junctions, cortical and spinal motor neurons (MNs), resulting in muscle paralysis. MNs are particularly vulnerable due to their long axons and high metabolic demands. ALS pathology involves impaired RNA metabolism, protein aggregation, cytoskeletal defects affecting organelle trafficking, and mitochondrial dysfunction, ultimately leading to neurodegeneration. Current treatments only marginally extend survival, calling for alternative therapies. We recently reported beneficial effects of electromagnetic alternating field stimulation (ACMS) on axonal regeneration and organelle motility in ALS. Here, we used iPSC-derived spinal MNs from familial ALS patients with FUS mutations to optimize magnetic field parameters regarding frequency, amplitude and duration. Using compartmentalized neuron cultures, we also tested different magnetic field orientations relative to axons. Following ACMS, we assessed axonal organelle trafficking and the nuclear DNA damage response (DDR) by live-cell imaging. Untreated ALS neurons showed clear defects in both assays. Exposure to magnetic AC sine waves above a threshold amplitude of 7 mT, applied perpendicular to the axonal plane with an optimal low frequency of 25 Hz, led to a sustained boost of mitochondrial and lysosomal motility for over one week and rehabilitation of DDR. These phenotypic improvements correlated with global transcriptomic changes revealed by RNA-sequencing. These phenotypic improvements correlated with global transcriptomic changes revealed by RNA-sequencing. While untreated mutant and wild-type neurons displayed different gene expression profiles, ACMS converged them to a novel, harmonized transcriptional state across all lines. Further analysis of differentially expressed genes (DEGs) yielded a conceptual model in which ACMS activates extracellular matrix (ECM)-mediated cell adhesion and neurotransmitter receptor, ion channel and kinase signaling pathways, ultimately modulating gene transcription and, thereby, motor- and microtubule-associated proteins. This model provides a framework for follow-up studies to experimentally validate the underlying mechanisms and assess the therapeutic potential of ACMS as an alternative biophysical treatment modality for neurodegenerative diseases.

Cells doi: 10.3390/cells15212002

Authors: Shiyan Su Jing Yang Meiyue Zhu Yunhua Duan Ruizhe He Banglan Luo Lei Tian Ye Li

Pancreatitis is a prevalent disorder of the digestive system, characterized by the rapid onset of pancreatic exudative edema and acinar cell necrosis. Clinically, the pathogenesis of pancreatitis is multifactorial and involves an intricate interplay among oxidative stress, dysregulated inflammatory signaling pathways, impaired autophagy, and aberrant apoptosis. Currently, no targeted disease-modifying pharmacotherapies are approved for clinical use in pancreatitis. In recent years, natural products have garnered increasing attention due to their inherent advantages, including chemical diversity, multi-target engagement, and favorable safety profiles, rendering them promising candidates for both preventive and therapeutic intervention in pancreatitis. Nevertheless, a comprehensive synthesis of current evidence regarding their mechanisms, bioactive constituents, and translational potential remains lacking. This review provides a rigorously categorized analysis of bioactive natural compounds (including alkaloids, quinones, terpenes, flavonoids, phenolics, and glycosides) and their molecular targets implicated in the prevention and treatment of pancreatitis. Furthermore, it critically evaluates emerging strategies for the rational engineering and optimization of plant-derived compounds, thereby offering a conceptual and mechanistic foundation for the development of novel therapeutics and innovative treatment paradigms for pancreatitis.

Cells doi: 10.3390/cells15212001

Authors: Sara Saleem Luke Gaughan Craig N. Robson

Hormone-driven cancers, including prostate, breast, endometrial and ovarian cancers, are a leading cause of cancer-related mortality. Endocrine therapies targeting androgen receptor and oestrogen receptor/progesterone receptor signalling have improved outcomes, but acquired resistance arising through genetic and epigenetic mechanisms that bypass receptor dependence remains a major challenge. BORIS (Brother of the Regulator of Imprinted Sites), a CTCF paralogue, is a germline-restricted regulator that becomes aberrantly reactivated in cancer through promoter hypomethylation, p53 loss, and loss of CTCF-mediated repression. Once expressed, BORIS occupies CTCF sites, displacing or co-occupying them with CTCF, and reprograms clustered sites into alternative transcriptional start sites through SRCAP-mediated H2A.Z deposition, activating germline genes that sustain proliferation, and therapeutic resistance. BORIS expression correlates with poor prognosis in ovarian and endometrial cancers, associates with ER/PR levels in breast cancer, and correlates with Gleason score in prostate cancer. BORIS represses AR signalling in ovarian cancer and inversely correlates with receptor expression in endometrial cancer. Its cancer-testis antigen properties position BORIS as an immunotherapy target, with preclinical vaccine efficacy demonstrated in breast cancer models. This review synthesises the disease-specific evidence positioning BORIS as a driver of transcriptional reprogramming and endocrine resistance, identifies current knowledge gaps, and discusses translational implications of targeting this pathway.

Cells doi: 10.3390/cells15212000

Authors: Zhenghao Wu Guqing Luo Jinbo Zhao Qingxun Zhou Hongjie Li Chihao Zhang Guangbo Wu Qiang Fan Xiaoliang Qi Haizhong Huo Jiayun Lin Meng Luo Lei Zheng

The liver lymphatic system plays an essential role during the progression of cirrhotic portal hypertension (PHT), featuring significantly enhanced lymphangiogenesis. However, the role of liver lymphatic functional drainage is understudied in cirrhosis. Therefore, this study aimed to investigate the effects of partial lymphatic vessel ligation (PLVL) of the liver on PHT. The rat model was established by common bile duct ligation (BDL), when PLVL was achieved via ligation of the portal and celiac lymph node. The severity of cirrhosis and PHT were evaluated, as well as macrophage infiltration and lymphangiogenesis. Bulk RNA-seq of liver was used to investigate the underlying mechanisms. Datasets of single-cell and bulk RNA-seq were obtained from the GEO database. Human dermal lymphatic endothelial cells (hLECs) were used for in vitro validation. Ligation of lymphatic drainage caused significant deterioration of liver cirrhosis and PHT in BDL rats. Our bulk RNA-seq suggested significant enrichment of vascular remodeling and angiogenesis-related pathways, as well as lipid metabolism-related biological functions in BDL rats after PLVL. Further analysis combined with public datasets identified several endothelial cell-specific upregulated genes in BDL and PLVL rats. LEC-derived upregulated genes were also identified from bulk RNA-seq of LECs isolated from mice liver, which were validated by incubating hLECs in TNF-α, LPS, TGF-β1. In the current study, we provide the first evidence that liver lymphatic drainage blockade aggravates BDL-induced cirrhotic PHT. LEC-associated secretory proteins might play a potential role in PHT progression, when hepatic inflammation and immune response are considered to be the essential regulatory mechanisms.

Cells doi: 10.3390/cells15211999

Authors: Darya A. Orlova Aleksandr S. Chernov Vitaly A. Kazakov Georgii B. Telegin Alexey A. Belogurov Anna A. Kudriaeva

Traumatic spinal cord injury (SCI) is believed to increase the risk of autoimmune diseases, including the autoimmune demyelinating disease multiple sclerosis (MS), in humans. The adaptive immune response against one of the key MS autoantigens, myelin basic protein (MBP), is elevated in mice and persists in patients with SCI even 10 years after the initial injury. Here, we show that β1i-enriched immunoproteasomes and PA28β subunits are elevated as early as 12 h post-SCI in DA rats, and all proteasome catalytic immunosubunits are highly abundant in the injured spinal cord of DA and SD rats 7–14 days post-SCI; these changes may therefore represent a primary event of the adaptive myelin-reactive immune response. Mogamulizumab bound a recombinant rat CCR4 N-terminal extracellular fragment, albeit less strongly than the corresponding human fragment, and treatment was associated with weaker CCR4-associated immunoreactivity in peripheral blood leukocyte lysates. Plasma samples from rats with SCI treated with mogamulizumab were characterized by lower levels of antibodies to MBP compared with those from the non-treated group, and by day 60 of the experiment, this discrepancy reached statistical significance. Compared with non-treated animals, rats treated with mogamulizumab showed a significantly decreased glial scar size and total area of cyst-like cavities. Thus, mogamulizumab therapy not only improves the morphological features of the glial scar but also suppresses the autoimmune response against myelin, and may offer a complementary immunological strategy for spinal cord protection following injury.

Cells doi: 10.3390/cells15211998

Authors: Mengyao Leng Xiuhong Weng Jingyi Luo Chenxing Hou Simin Wang Zhiqiang Li Bo Cheng

Glioma is the most prevalent and aggressive brain tumor with high recurrence rates and poor survival. Porphyromonas gingivalis (P. gingivalis), a major periodontal pathogen, has emerged as a potential contributor to neurological diseases. However, whether P. gingivalis is present in glioma tissues and directly promotes malignant progression remains unclear. Here, immunohistochemistry the detected gingipain in 11 of 13 human glioma specimens, while fluorescence in situ hybridization further detected P. gingivalis-targeted 16S rRNA signals in 3 of 13 specimens. These findings support the presence of P. gingivalis-associated signals in human glioma tissues. In cellular models, P. gingivalis invaded glioblastoma cells and promoted cell proliferation, migration, and invasion through induction of mesenchymal transition and extracellular matrix remodeling, thereby accelerating glioma progression. In a mouse xenograft model, P. gingivalis inoculation enhanced tumor growth and was associated with changes in proliferation, mesenchymal transition, and ECM remodeling-related markers. Mechanistically, P. gingivalis activated the PI3K/AKT/mTOR signaling pathway, while pharmacological AKT inhibition (AKTi-1/2) effectively suppressed these tumor-promoting effects. Metronidazole treatment also reduced tumor growth in vivo. Collectively, these findings identify P. gingivalis as a potential microbial contributor to glioma malignant progression and support further investigation of host–microbe interactions and AKT signaling as potential avenues for therapeutic research.

Cells doi: 10.3390/cells15211996

Authors: Asma Mahdi Arwa Aldaalis Georges Chehade Eva On-Chai Lau Hend Salah Fayed Ayman Al Haj Zen Mohamed Aittaleb Maisa Ismael Alkailani Fadel Tissir

In humans, low expression of DIAPH3 is associated with poor prognosis in glioblastoma patients. In mice, DIAPH3 loss induces aberrant mitosis, chromosomal instability, and cell death; whereas the combined loss of DIAPH3 and p53 causes high-grade diffuse glioma and accelerates tumor onset. In this study, we used Diaph3/p53 double conditional knockout mice (dcKO) and ultra-high-resolution MRI to detect and characterize the development and progression of glioma. We transcriptionally profiled early and advanced gliomas and identified stage-dependent alterations in molecular signatures as the tumor progresses. Notably, genetic pathways underlying self-renewal and extracellular matrix (ECM) interactions were upregulated at the expense of neural activity signature, which was severely shrunk. CKAP2L was identified as a differentially expressed gene at both early and advanced tumor stages in the dcKO mice. A subsequent siRNA screen to identify genes involved in proliferation and migration of glioblastoma cells identified CKAP2L as a top hit. Functional studies showed that CKAP2L depletion reduced proliferation and migration and induced cell accumulation in G2/M phase of the cell cycle. We also used data mining and thoroughly characterized the relationship between CKAP2L expression and glioma development. We found that CKAP2L expression correlates with proliferative and progenitor-associated markers, pointing to a potential role in maintaining highly proliferative and progenitor-like tumor cell state. We also identified a differentially methylated CpG site in the CKAP2L promoter that was associated with changes in gene expression across glioma datasets. Overall, our findings show that CKAP2L may contribute to glioma progression, help distinguish tumor grade, and represent a potential therapeutic target in glioblastoma treatment.

Cells doi: 10.3390/cells15211997

Authors: Itika Arora Suvasmita Rath Qazi Mohammad Sajid Jamal Ahmed Aljohani Abid Hamid Ahmed Abu-Zaid Mohammad Imran Khan Ahmed Yaqinuddin

PTEN loss activates PI3K–AKT signaling, and mTORC1 stimulates pyrimidine synthesis through S6K1-mediated CAD phosphorylation. This exploratory public-data analysis tested whether PTEN deletion in primary prostate cancer is associated with CAD, mTORC1 and pyrimidine-synthesis expression, and whether these survive protein-level and confounder analyses. In 485 primary tumors from The Cancer Genome Atlas Prostate Adenocarcinoma (TCGA-PRAD; 159 PTEN-deleted), deletion was associated with higher CAD mRNA (Hedges’ g = 0.57) and higher Hallmark mTORC1 scores (g = 0.54). In 342 tumors with reverse-phase protein-array data, deletion was accompanied by lower PTEN protein and higher AKT Ser473 phosphorylation but not by a higher pre-specified mTORC1 phospho-output composite (g = −0.02), itself uncorrelated with the transcriptional score. In a pre-specified model adding purity, TP53, RB1, ERG fusion and batch (n = 276), the PTEN–CAD association was absent (β = −0.002; p = 0.96), whereas the mTORC1-score association persisted. In a post hoc decomposition, ERG fusion, which co-occurs with PTEN deletion (odds ratio 3.6), changed the coefficient most. Across eight cohorts, a small PTEN mRNA–CAD association (g = 0.23) was attenuated to 0.14 after ERG adjustment, with overlapping confidence intervals (Hartung–Knapp p = 0.071). Bulk-tumor pathway signatures and PTEN–CAD associations need protein-level and ERG-aware interpretation.

Cells doi: 10.3390/cells15211995

Authors: Zbigniew Krasiński Giuseppe Maria Raffa Mariusz Kowalewski Beata Krasińska Calogera Pisano Stefano Cannata Piotr Suwalski Tomasz Urbanowicz

Atherosclerotic carotid plaques are shaped by years of interaction between systemic risk factors, hematopoietic conditioning, vascular injury, and local immune responses, yet conventional assessment captures only a limited snapshot of this biological history. This review introduces the concept of the “immunological biography” of the carotid plaque and examines how trained innate immunity, macrophage state and defective resolution, adaptive immune memory, T- and B-cell clonal selection, and spatially organized immune niches contribute to plaque heterogeneity and vulnerability. The literature was identified through a structured PubMed search complemented by reference screening, with emphasis on human carotid tissue, paired blood–plaque studies, immune-repertoire analyses, and single-cell and spatial technologies. Current evidence indicates that immune-cell abundance alone incompletely characterizes plaque biology: functional state, antigen experience, clonality, tissue localization, and interactions with vascular and stromal cells provide additional levels of biological information. We propose the Carotid Plaque Immune Phenotype (CPIP) as an integrative framework linking systemic immune history with cellular state, clonal architecture, spatial organization, and clinically relevant plaque features. Combining circulating biomarkers with tissue-resolved immune profiling may ultimately improve biological phenotyping of carotid atherosclerosis and provide a framework for prospective studies linking plaque immune architecture with cerebrovascular events.

Cells doi: 10.3390/cells15211994

Authors: Engelinah Dikeledi Macamo Pragalathan Naidoo Sibusiso Senzani Khethiwe Nomcebo Bhengu Sayed Shakeel Kader Zilungile Lynette Mkhize-Kwitshana

There is a rapid increase in early onset CRC worldwide. This incidence is profoundly influenced by biological aging in the colon, yet the underlying mechanisms remain incompletely defined. This review critically evaluates evidence supporting proposed interactions among telomere dynamics, sirtuins, and mitochondrial biomarkers during colon aging and as CRC risk predictors, while highlighting areas where mechanistic validation remains limited. This insight paves the way for novel early-detection biomarkers, personalized preventive approaches, and treatments designed to directly interfere with aging mechanisms and CRC risk.

Cells doi: 10.3390/cells15211993

Authors: Chenhui Ren Lu Wei

Matrix metalloproteinase-2 (Mmp2, also known as gelatinase A) is a zinc-dependent endopeptidase, and mutations in human MMP2 can lead to some human diseases. However, the underlying function and mechanism of mmp2 during eye development are still unclear. The present study generated mmp2 crispant zebrafish by CRISPR/Cas9 system. Whole-mount in situ hybridization (WISH), immunofluorescence, qRT-PCR, and transcriptomic and metabolomic analyses were performed. Results showed that mmp2 knockdown led to small eyes at 48–96 hpf (hours post-fertilization). WISH and qRT-PCR showed that mmp2 knockdown at 72 hpf (hours post-fertilization) significantly decreased the expression of cone-specific markers (opn1sw1, opn1sw2, opn1mw1, and opn1lw1), the rod-specific marker rhodopsin, retinal cell-specific markers (gnat1 and gnat2), and lens-specific markers (crybgx and crygm2c) in the retina. Immunofluorescence showed increased apoptosis (TUNEL+ cells), ectopic localization of the neuronal differentiation marker HuC/D at the ciliary marginal zone, and decreased photoreceptor cells (Zpr1) and photoreceptor (RCVRN) fluorescence. Transcriptomic analysis indicated that 520 differentially expressed genes were enriched in phototransduction and retinol metabolism. Metabolomics analysis indicated that mmp2 knockdown led to abnormal lipid metabolism and amino acid metabolism. Integrated omics analysis revealed shared pathways involving PPAR signaling and nucleotide sugar metabolism, while correlation analysis highlighted strong associations between eye-development-related genes (e.g., gnat1, gnat2, opn1sw1) and specific metabolites such as 2-palmitoyl-rac-glycerol and 1-hexadecyl-sn-glycero-3-phosphocholine. Collectively, these findings demonstrate that Mmp2 is essential for normal eye development in zebrafish, and its deficiency disrupts retinal cell survival, differentiation, and metabolic homeostasis, providing new insights into the molecular mechanisms underlying Mmp2-associated ocular abnormalities.

Cells doi: 10.3390/cells15211992

Authors: Weichao Ling Meijing Song Xinlin Li Yu Li Yuhao An Zhidong Chen Zhijun Ruan Yang Zhou Linlin Lu Zigang Li Rui Wang

Pachymic acid (PA), a major bioactive component of Poria cocos, suppresses colorectal cancer (CRC) proliferation and invasion by disrupting glycolysis. However, its antitumor activity and underlying mechanisms in vitro and in vivo remain unclear. RNA-binding proteins (RBPs) are key post-transcriptional regulators in CRC, but their mechanisms of action and therapeutic potential remain poorly understood. Here, we employed limited proteolysis-mass spectrometry (LiP-MS), a target deconvolution approach that does not require prior chemical modification, to profile changes in protein conformation within complex proteomes upon PA treatment in cells. Our findings revealed that RBMX (RNA-binding motif protein X-linked) acts as a target of PA and undergoes concentration-dependent conformational changes, as supported by binding validation and functional assays performed in vitro and in vivo. Furthermore, proximity-labeling mass spectrometry (PL-MS) analysis revealed that RBMX interacts with heterogeneous nuclear ribonucleoprotein C (hnRNPC). Functional assays demonstrated that PA treatment induces RBMX-dependent degradation of hnRNPC, thereby suppressing CRC cell proliferation and tumor growth. In summary, this study identifies RBMX as a molecular target of PA through LiP-MS, providing new insights into the mechanisms underlying PA-mediated CRC suppression and highlighting the RBMX–hnRNPC axis as an important mediator of its antitumor effects.

Cells doi: 10.3390/cells15211991

Authors: Alejandra Varela Samar Sayedyahossein Rithwik Ramachandran Lina Dagnino

Kindlins are essential for integrin activation and mechanoresponses. In the epidermis, inherited loss-of-function of the FERMT1 gene, which encodes Kindlin-1, increases the risk of developing cutaneous squamous cell carcinoma (cSCC). Notably, the role of Kindlin-1 in epidermal tumours has remained virtually unexplored. In this study, we analyzed the consequences of FERMT1 inactivation in a cSCC cell model. Kindlin-1-depleted cells exhibited decreased number and size of focal adhesions, as well as alterations in actin stress fibres and the F-actin perinuclear cap. Significantly, FERMT1 inactivation also resulted in reduced nuclear volume and total surface area. These abnormalities were not ameliorated by exogenous expression of Kindlin-2 or a Kindlin-1 mutant incapable of binding to integrins. In contrast, nuclear dimensions in Kindlin-1-deficient cells were restored in the presence of a constitutively active RhoA protein. These findings are consistent with the concept that Kindlin-1 plays an essential role mediating integrin signalling and RhoA activation to modulate perinuclear actin filaments and nuclear morphology.

Cells doi: 10.3390/cells15211990

Authors: Erica Candela Giovanna Forte Matteo Iacoviello Antonia Lucia Buonadonna Anna Maria Valentini Martina Lepore Signorile Paola Sanese Katia De Marco Andrea Manghisi Nicoletta Maria Tutino Emanuele Tatò Candida Fasano Vittoria Disciglio Valentina Grossi Cristiano Simone

Missense variants in PMS2 remain difficult to interpret in Lynch Syndrome (LS), particularly when clinical evidence is limited. We investigated the PMS2, NM_000535.7: c.2103T>A; p.His701Gln (p.H701Q) variant, currently classified as a variant of uncertain significance (VUS), identified in a 71-year-old patient with small intestinal adenocarcinoma. To clarify its clinical significance, we combined clinical, in silico, and functional analyses. Computational predictions consistently pointed to a deleterious effect, with AlphaMissense assigning a highly damaging score and most independent predictors supporting the same interpretation. H701Q affects the highly conserved C-terminal region of PMS2, close to the functionally critical Glu705 residue. In HEK-293T cells, PMS2-H701Q showed markedly reduced protein levels compared with wild-type PMS2, supporting an effect on protein stability. Co-immunoprecipitation experiments further showed a substantial reduction in the interaction between PMS2-H701Q and MLH1, indicating impaired MutLα complex formation. Both effects were comparable to those observed for the pathogenic PMS2-E705K control. Overall, the agreement between computational and functional findings provides compelling evidence for a deleterious effect of H701Q and strengthens its clinical interpretation. Collectively, these findings demonstrate the utility of functional evidence under the ClinGen-InSiGHT PMS2-specific ACMG/AMP variant classification guidelines and support reclassifying the PMS2, NM_000535.7: c.2103T>A; p.His701Gln (p.H701Q) variant from a Variant of Uncertain Significance (VUS) to likely pathogenic.

Cells doi: 10.3390/cells15211988

Authors: Xin Xu Guangyu Wu

G protein-coupled receptors (GPCRs) constitute the largest family of membrane receptors that regulate a vast variety of cellular functions and serve as important therapeutic targets. A fundamental but poorly addressed question in GPCR biology is how individual receptors are efficiently delivered to their functional destinations following synthesis in the endoplasmic reticulum (ER). Ufmylation is a ubiquitination-like post-translational modification in which ubiquitin-fold modifier 1 is conjugated to target proteins. Compared with ubiquitination, the physiological and pathological significance of ufmylation is much less defined. Recent studies, however, have revealed crucial functions of the ufmylation system in maintaining ER homeostasis and directing GPCR export from the ER. In this review, we succinctly summarize recent advances in understanding the ufmylation system, discuss its role in the biosynthetic transport and segregation of nascent GPCRs at ER-derived COPII vesicles, and explore the therapeutic potential of targeting ufmylation-mediated GPCR trafficking.

Cells doi: 10.3390/cells15211989

Authors: Chu-Che Lee Yong-Syuan Chen Shun-Fa Yang Chu-Liang Lin Yi-Hsien Hsieh Chun-Yi Chuang

Background: Nasopharyngeal carcinoma (NPC) is a highly metastatic epithelial malignancy with limited therapeutic options for advanced disease. Timosaponin AIII (TSAIII), a natural steroidal saponin, exhibits antitumor activity; however, its effects on NPC progression and the molecular mechanisms remain unclear. Method: Cell viability, proliferation, apoptosis, migration, and invasion were evaluated using cell growth, colony formation, Annexin V/PI, and Transwell assays. Human chemokine array and The Cancer Genome Atlas (TCGA) analyses identified midkine (MDK) as a potential molecular target of TSAIII and its clinical relevance in NPC. Western blotting, qRT-PCR, immunofluorescence, and chromatin immunoprecipitation (ChIP) assays were used to investigate the underlying signaling mechanisms. Antimetastatic activity was further evaluated in C.B17-SCID mice. Results: TSAIII suppressed NPC cell migration and invasion with minimal cytotoxicity, accompanied by reduced the expression of p-AKT (Ser473), estrogen receptor β(ERβ) nuclear localization, and MDK expression. Wortmannin enhanced these effects, while TSAIII attenuated latent membrane protein 1 (LMP1)-induced signaling. In vivo, TSAIII significantly reduced lung metastasis without apparent toxicity. Conclusions: TSAIII suppresses NPC metastasis by targeting the LMP1/p-AKT/ERβ/MDK signaling axis, supporting its potential as an antimetastatic therapeutic candidate for NPC.

Cells doi: 10.3390/cells15211987

Authors: Nuray Üremiş Muhammed Mehdi Üremiş

Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) are considered promising biological carriers for biomolecule and drug delivery because of their inherent biological activities, biocompatibility, and capacity to carry a variety of therapeutic cargoes. However, the limited cargo-loading capacity of native EVs, their inefficient delivery to target cells, intracellular trafficking barriers, and limitations in the functional release of their cargo restrict their therapeutic applications. EV engineering encompasses a range of approaches to overcome these limitations, including donor cell modification, endogenous cargo loading and active sorting mechanisms, post-isolation loading methods, surface engineering, and EV–lipid-based hybrid systems. In this review, engineered MSC-EVs are evaluated in terms of cargo loading, targeting, cellular uptake, intracellular trafficking, and functional cargo delivery. We also compare the loading strategies used for nucleic acids, proteins, enzymes, and small molecules, in terms of their advantages, their limitations, and their effect on EV biological activity. The therapeutic applications of MSC-EV-based systems in cardiovascular diseases, wound healing, neurological and musculoskeletal disorders, inflammatory diseases, and cancer are evaluated. The central conclusion of this review is that the therapeutic efficacy of MSC-EVs depends not only on the amount of cargo delivered but also on its delivery to the appropriate target cells, intracellular targeting, and preservation of its biological activity. Accordingly, future EV engineering strategies should aim to simultaneously improve cargo loading, disease-specific targeting, local retention, and cargo stability. Progress on these four fronts is what would make MSC-EVs predictable enough for clinical use.

Cells doi: 10.3390/cells15211986

Authors: Anita Steib Katalin Rozmer Zoltán Sándor Éva Szőke Dávid Semjén Béla Kajtár Krisztina Pohóczky Zsuzsanna Helyes

Transient receptor potential ankyrin 1 (TRPA1) and vanilloid 1 (TRPV1) have been implicated in cancer biology, but their functional relevance in prostate adenocarcinoma (PAC) remains unclear. Therefore, we characterized TRPA1/TRPV1 expression and function in benign prostatic hyperplasia (BPH), PAC tissues, and PAC cell lines with different characteristics and examined the effects of their agonists on cancer-cell viability. TRPA1 and TRPV1 transcripts were assessed by RNAscope in situ hybridization and RT-qPCR, channel activity by Fura-2-based Ca2+ imaging, and viability/proliferation by ATP-based luminescence and automated cell counting. TRPA1 and/or TRPV1 mRNAs were detected in PAC tissues and cell lines, but selective stimulation with allyl-isothiocyanate (AITC) or capsaicin did not evoke Ca2+ influx, indicating no detectable functional channel activity under our experimental conditions. Non-selective TRPA1 activators induced Ca2+ influx responses that were not inhibited by the receptor antagonist A-967079. AITC concentration dependently reduced viability and proliferation of the androgen-sensitive 22Rv1 and LNCaP cells, and these effects were not prevented by TRPA1 antagonists. Capsaicin had no effect at TRPV1-selective concentrations. Thus, transcript expression does not predict functional TRPA1/TRPV1 activity in PAC cells, and AITC reduces PAC-cell viability through TRPA1-independent mechanisms.

Cells doi: 10.3390/cells15211985

Authors: Ning Liu Xiaowen Chen Hailong Zhao Bo He

Protein tyrosine sulfation (PTS), an important post-translational modification (PTM), has garnered increasing attention in life sciences in recent years. Following classical and emerging PTMs such as phosphorylation, lactylation, acetylation, and palmitoylation, PTS has now emerged as a novel entry point in disease mechanistic studies. As an evolutionarily conserved modification ubiquitously present in organisms, PTS acts as a core molecular switch that modulates fundamental biological processes via specific conjugation of sulfate groups to tyrosine residues of target proteins. Accumulating evidence has revealed that PTS modifies numerous pivotal functional proteins, such as chemokine receptors, histones, and coagulation factors, and participates in multiple physiological and pathological events, including hormonal modulation, gene transcription, hemostasis, inflammatory responses, cancer progression, and viral infection. Consequently, deeper insight into the mechanisms of PTS and its targeted intervention holds promise for novel therapeutic strategies for related diseases. Nevertheless, the molecular regulatory network of PTS remains poorly understood, which is largely attributed to the limitations of current techniques for the efficient preparation and accurate detection of PTS-modified proteins. This review systematically summarizes the latest advances in the biological functions and regulatory mechanisms of PTS, aiming to provide a theoretical reference for subsequent fundamental research and offer potential therapeutic targets for the clinical treatment of PTS-related diseases.

Cells doi: 10.3390/cells15211984

Authors: Maddox Harrison Nicolaos Achilleos Evangelia Tsiani

Insulin-sensitive tissues, including skeletal muscle, adipose tissue, and liver, are central regulators of systemic glucose and lipid homeostasis. Insulin signaling disruption in these tissues is a key pathological feature underlying insulin resistance and contributes to the development of metabolic disorders such as type 2 diabetes mellitus (T2DM), obesity, and liver disease. Increasing evidence suggests that berberine (BBR), a naturally occurring isoquinoline alkaloid derived from Berberis species, attenuates insulin resistance and exhibits antidiabetic, lipid-lowering, and anti-inflammatory properties across multiple metabolic tissues. This review analyzes current in vitro, in vivo, and clinical data examining the effects of BBR in insulin-sensitive tissues. In skeletal muscle, BBR promotes glucose consumption while in adipose tissue, it attenuates adipogenesis, lipid accumulation and inflammation. In hepatic tissue, BBR suppresses gluconeogenesis, improves lipid metabolism and reduces hepatic steatosis. Clinical trials report reductions in fasting glucose, serum lipids, visceral adiposity, and hepatic fat content. Collectively, current evidence indicates that BBR exerts multi-tissue insulin-sensitizing effects with therapeutic potential for metabolic disease. However, mechanistic human data directly linking tissue-specific actions remain limited. Future clinical studies incorporating tissue-specific metabolic assessments, dose–response analyses, metabolomic profiling, and consideration of sex- and age-related variability are warranted to fully elucidate its clinical efficacy.

Cells doi: 10.3390/cells15211983

Authors: Jia-Yi Zhang Xin-Yi Tong Run-Min Tan Zhi-Lin Yu Jian-Ming Ouyang

Magnesium ammonium phosphate (MAP) stones feature a markedly high recurrence rate. The inflammatory microenvironment contributes to MAP crystal-induced urothelial injury, yet the specific regulatory mechanisms remain unclear. In this study, MAP crystals were synthesized, and pharmacological intervention with the ROS scavenger N-acetylcysteine (NAC), the CTSB inhibitor CA-074 Me, and the NLRP3 inhibitor MCC950 was used to explore the signaling pathway through which MAP crystals induce inflammatory responses; an inflammatory model of SV-HUC-1 urothelial cells was established using lipopolysaccharides (LPSs) to observe the regulation of key molecules in the inflammatory signaling pathway and its effects on MAP crystal adhesion and cellular injury. Results showed that MAP crystals elevated intracellular ROS levels, aggravated lysosomal membrane permeabilization, triggered CTSB leakage, and activated the NLRP3 inflammasome pathway. In the LPS-induced inflammatory microenvironment, MAP crystals exacerbated cellular inflammatory responses and cell injury. Meanwhile, the expression of adhesion-related proteins, including OPN, CD44, and ICAM-1, was further upregulated, accompanied by aggravated MAP crystal adhesion. In summary, MAP crystals induced inflammation in SV-HUC-1 cells via the ROS–CTSB–NLRP3 axis, and the LPS-induced inflammatory microenvironment further amplified cellular inflammation, cell damage, and crystal adhesion. This study provides theoretical evidence supporting the pro-inflammatory effect of the inflammatory microenvironment on MAP stone formation.

Cells doi: 10.3390/cells15211982

Authors: Raquel Gomez Junco Lobo Akshay Uttarkar Jeannette Lager Aditi Bhargava

Maternal stressors during pregnancy are linked to adverse birth outcomes, but the biochemical pathways involved remain poorly defined. We previously reported that the human placenta buffers the fetus from several effects of maternal perceived stress in a cohort of term pregnancies. Here, using the same cohort and no additional participants, we analyze a broader steroid panel and, for the first time, a targeted bile acid metabolomic panel across four maternal–fetal compartments: maternal plasma, placental tissue, cord plasma, and maternal hair. Nulliparous women were recruited during the third trimester at UCSF Mission Bay Hospital; perceived stress was measured using the 10-item Perceived Stress Scale (PSS) and retained as a continuous variable for the primary analysis from 17 women. Of 212 analyte compartment pairs evaluated, 38 metabolites met a predefined detectability criterion and were tested for association with PSS and sociodemographic scores using Kendall’s tau b, with permutation p-values, bootstrap confidence intervals, and Benjamini–Hochberg correction for multiple comparisons. No association survived multiple testing corrections. In exploratory, uncorrected comparisons (p < 0.05), three metabolites were nominally lower in a cohort-derived, higher-PSS group: placental estrone (E1), cord plasma glycochenodeoxycholic acid (GCDCA), and maternal plasma ursodeoxycholic acid (UDCA). Nearly 47.1% of women had gestational diabetes; 20-hydroxyprogesterone (20-OH-PROG) in maternal plasma, dihydroprogesterone (DHP), and pregnenolone (PELN) in placental tissues were nominally decreased in women who developed gestational diabetes compared to women who did not develop gestational diabetes. Maternal hair was not a good compartment for detection of bile acid metabolites. Despite a wide range of perceived stress/sociodemographic scores and a substantial burden of maternal clinical comorbidities in this cohort, all pregnancies resulted in favorable neonatal outcomes at delivery. These findings support the hypothesis that the placenta functions as a buffering organ that protects the developing fetus against broad insults while remaining susceptible to more selective disruptions.

Cells doi: 10.3390/cells15211981

Authors: Yueying Zhang Keying Li Xixi Yang Wei Zhang Mingtao Ma Nanjian Luo

Transcripts annotated as non-coding can yield both an RNA molecule and a translated product, complicating causal attribution in skeletal muscle. Three distinct claims require different evidence: a product exists, the nominated RNA generated it, and the product drives a defined phenotype. We examine them in myogenesis and regeneration using a qualitative, two-axis evidence map. The T axis records translation and RNA-of-origin evidence, whereas the F axis records experiments separating RNA-dependent from product-dependent effects for a stated model and endpoint. Endogenous coding-selective experiments support qualified product-dependent assignments for LINC00961–SPAR and MTLN/MPM. Linc-RAM/MRLN demonstrates biologically active RNA and peptide outputs, but not that both cause the same phenotype. Among the selected circRNA systems, circZNF609, circFAM188B, circNEB, and circSIK2 retain unresolved source and/or functional attribution, and none meets both T4 and F4. Rather than ranking study quality or assuming transfer across species and myogenic stages, the map identifies the experiment still needed to establish causality.

Cells doi: 10.3390/cells15211980

Authors: Yaolong Song Yingchao Li Mingshuai Chen Chenyang Shi Qirui Zang Wanjing Jin Qian Ma Binhuan Ma Panpan Tong Shicheng Wan Aili Aierken Mengfei Zhang

Bovine mastitis is a major infectious disease in dairy cattle, and the increasing prevalence of antimicrobial-resistant pathogens has created an urgent need for the development of effective antimicrobial agents. In this study, bovine enteric β-defensin (EBD) from Xinjiang Brown Cattle was cloned, expressed, and preliminarily evaluated for its antibacterial and protective effects against Enterococcus faecalis-induced mastitis. Bioinformatic analysis identified EBD as a secreted protein belonging to the bovine β-defensin family. The recombinant PCDH-EBD construct was successfully transfected into 293T cells, resulting in markedly increased EBD mRNA expression and detectable recombinant EBD protein expression, as confirmed by SDS-PAGE and Western blotting. In vitro agar diffusion assays showed that the EBD-containing preparation exhibited inhibitory activity against E. faecalis. In a murine mastitis model, EBD treatment significantly attenuated mammary histopathological lesions and reduced the expression of Mpo, Tnf, and Il6 compared with the mastitis group. 16S rRNA sequencing of mammary gland tissues showed that EBD treatment was associated with a reduced relative abundance of Enterococcus and partial restoration of microbial diversity. Transcriptomic analysis further revealed that EBD treatment was associated with alterations in NOD-like receptor signaling- and phagosome-related pathways, suggesting that these processes may contribute to its protective effects. Collectively, these findings indicate that enteric β-defensin EBD from Xinjiang Brown Cattle possesses antibacterial and anti-inflammatory potential, making it a candidate for developing antimicrobial agents against bovine mastitis.

Cells doi: 10.3390/cells15211979

Authors: Katarzyna Herbetko Aleksandra Herbetko Sara Suwała Jakub Czarny Julia Rudno-Rudzińska Olga Milbrandt Julita Kulbacka

Colorectal cancer (CRC) remains one of the most common malignancies worldwide and a leading cause of cancer-related mortality. Recent advances in CRC molecular biology have focused on biomarkers for early detection, prognostic stratification, treatment selection, and disease monitoring. Clinically validated biomarkers, including microsatellite instability (MSI), RAS and BRAF mutations, HER2 amplification, and carcinoembryonic antigen (CEA), guide targeted therapy and predict treatment response. However, tumor heterogeneity, clonal evolution, and microenvironmental influences limit the predictive value of single biomarkers. Recent molecular diagnostics therefore increasingly focus on dynamic and minimally invasive approaches. Circulating tumor DNA (ctDNA) is a promising biomarker for detecting minimal residual disease, monitoring recurrence, and assessing tumor evolution in real time. Multi-omics profiling, spatial transcriptomics, and artificial intelligence further expand tumor characterization except for conventional genomic alterations. Our review integrates common CRC biomarkers with developing liquid biopsy, multi-omics, spatial transcriptomics, and artificial intelligence approaches across early detection, prognosis, treatment selection, and longitudinal monitoring. Future biomarker strategies will likely rely on integrated molecular models combined with longitudinal liquid-biopsy assessment. However, most new biomarkers remain at the investigational stage, and assay heterogeneity, limited standardization, and insufficient prospective clinical validation currently restrict their routine clinical use.

Cells doi: 10.3390/cells15211978

Authors: Saima Usman William Andrew Yeudall Ahmad Jamal Muy-Teck Teh Ahmad Waseem

Cancer stem cells (CSCs) are a small population of cancer cells that endow unique traits of self-renewal, differentiation, tumorigenicity, quiescence and therapeutic resistance, ultimately leading to cancer relapse and poor prognosis. Conventional chemo- and radiotherapies effectively destroy rapidly dividing cancer cells; however, CSCs are largely refractory to these treatments due to special characteristics that include metabolic plasticity, low reactive oxygen species (ROS) levels, apoptosis inhibition and modified DNA repair systems, effectively hijacking the tumour microenvironment and evading host immune surveillance. This review summarises the potential CSC diagnostic methods that can be used in routine clinical investigation for tailoring personalised treatment plans both pre- and post-surgery. In this way, a CSC gene signature, reactivity and load can be estimated and, by using artificial intelligence and machine learning algorithms in a personalised therapeutic regimen, the most effective treatment response can be predicted. Furthermore, we propose incorporation of CSC-targeted therapies in conjugation with conventional chemo- and radiotherapies to eradicate CSCs, thereby eliminating the risk of relapse and therapy failure in cancer patients. This review also describes multiple potential molecular targets that may be used to target CSCs and emphasises the need for further evaluation, in order that they may be applied clinically.

Cells doi: 10.3390/cells15211977

Authors: Ching-Yi Lin Alex Nelson Jason Lin Wayne Lee Sidney Bihrer Yu-Shang Lee

Spinal cord injury (SCI) affects millions of people worldwide, resulting in persistent direct and secondary effects including but not limited to motor, sensory, and autonomic dysfunction. The widespread nature of this condition, as well as its detrimental effects, makes recovery following SCI an important clinical problem. One primary issue identified in SCI among other neurological conditions is fibrotic scarring, as its dynamic nature provides both beneficial and detrimental effects over its developmental course. Initially, early scar formation contributes to wound closure and tissue stabilization, but chronic, persistent fibrosis later interferes with axonal growth and other neural repair mechanisms. The scar itself is highly dynamic, consisting of components including collagens, fibronectin, proteoglycans, and laminins, all of which have diverse localization and functionality throughout scar maturation. Its formation is regulated by various signaling mechanisms, including inflammatory signaling, profibrotic pathways, vascular responses, interactions between fibroblasts and neighboring cells, and changes in the specific mechanical environment of the injury site. This review focuses primarily on SCI, with findings from additional central nervous system injury and disease models used to provide a holistic, clinically relevant overview of fibrotic scar organization, profibrotic signaling, and potential therapeutic targets.

Cells doi: 10.3390/cells15211976

Authors: Ilya Vyatchin Vyacheslav Dyachuk

This study investigated the formation of the contractile apparatus in bivalve mollusk larvae. Elucidation of the molecular aspects of larval development in bivalves is significantly complicated by the lack of antibodies that can selectively label proteins and their structures. To address this issue, polyclonal antibodies were produced against main proteins of bivalve muscles. Immunohistochemistry was used to obtain high-quality confocal images of muscles in bivalve larvae to describe the inclusion of main muscle proteins in the contractile apparatus during the development of the oyster Magallana gigas. The appearance of these proteins in muscle cells was not uniform and, surprisingly, the leading protein in the formation of muscle structures, including striations, was twitchin. The findings obtained support the hypothesis that this protein, similar to vertebrate titin (a protein from the same family), may play an essential role in the assembly of the contractile apparatus. Like another protein detected during this study, myorod, twitchin is believed to be involved in the formation of the catch state (an energy-efficient state of molluskan smooth muscles). The strong evidence for the presence of these proteins within the striated muscles in the larval stage of bivalves raises questions about the origin of this important evolutionary adaptation.

Cells doi: 10.3390/cells15211975

Authors: Shontol Bhowmic Pritha Roy Choudhury Prodipto Das Saptak Banerjee Jayanta Chakrabarti Subir Kumar Juin Sweta Ghosh

Resistance to chemotherapy continues to be a significant challenge for cancer treatment and a contributing factor to treatment failures and tumor progression. Recently, the gut microbiome has emerged as an important modulator of the response to chemotherapy. Numerous pre-clinical and clinical studies have demonstrated a direct association between gut microbiota composition and therapy effectiveness in many cancer types, such as colorectal, breast, pancreatic, gastric, and lung cancers, melanoma, and hematological cancers. Gut microbiota effectively regulate the efficiency of chemotherapeutic agents in various ways, either by biotransformation of chemotherapeutic agents or modulation of host immunity. In addition, microbial metabolites such as short-chain fatty acids, secondary bile acids, and tryptophan metabolites are also involved in the regulation of chemotherapy response through modulating apoptosis, immunity, and epithelial–mesenchymal transition. Finally, dysbiosis induced by chemotherapy and antibiotics has also been reported to reduce the effectiveness of therapy. This review will emphasize the ways in which gut microbiota can modulate chemotherapeutic response and induce chemoresistance. Understanding the interactions between chemotherapeutic drugs and the gut microbiota can help to enhance the efficiency of chemotherapy and find more appropriate personalized treatments.

Cells doi: 10.3390/cells15211974

Authors: Adnan Amin Wajid Zaman

Plants rarely encounter environmental constraints in isolation. Water deficit, temperature extremes, salinity, flooding, excessive irradiance, nutrient imbalance, herbivory, and microbial infection frequently overlap in time and space, creating composite cellular states that cannot be inferred by adding single-stress responses. This review develops an organelle-centred framework for such integration. We synthesize evidence that the plasma membrane and cell wall initiate partially shared Ca2+, reactive oxygen species (ROS), kinase, lipid, and electrical signals, whereas chloroplasts, mitochondria, peroxisomes, the endoplasmic reticulum (ER), and the vacuolar system transform these inputs into compartment-specific retrograde information. We distinguish experimentally supported plant contact-site functions from the hypothesis that contact-site remodeling encodes combined biotic–abiotic stress. We further examine how ABA-, SA-, JA-, and ET-centered hormonal circuits intersect with organellar signals, how the unfolded protein response and selective autophagy set stress thresholds, and how systemic signaling and chromatin-associated memory alter subsequent responses. Because most molecular memory evidence derives from single-stress acclimation or immune priming, persistent memory after combined stress remains an open question. An interface-to-memory model is proposed in which stress combinations are encoded through the timing, amplitude, localization, and persistence of signals, with implications for organelle-resolved experiments and field validation.

Cells doi: 10.3390/cells15211973

Authors: Zahra Enteshari Najafabadi Ireneusz Ryszkiel Hossein Baharvand Amir Sadeghi Agata Stanek Anna Meyfour

Gastric organoids are advanced three-dimensional models generated from adult or pluripotent stem cells. By reproducing key structural, cellular, and functional features of gastric tissue, they overcome several limitations of conventional two-dimensional cultures and animal models. Gastric organoids provide physiologically relevant platforms for studying stomach development, cellular differentiation, and the molecular mechanisms underlying gastric diseases. This review summarizes the principal cell sources and protocols used to generate gastric organoids and discusses recent advances in culture systems, including two-dimensional organoid-derived monolayers, air–liquid interface cultures, co-culture systems, and organoid-on-a-chip technologies. Particular attention is given to the application of gastric organoids in modeling Helicobacter pylori infection, SARS-CoV-2 infection, and gastric cancer, as well as in evaluating therapeutic responses. Their roles in developmental biology, the investigation of congenital disorders, drug screening, potential precision-medicine application, biobanking, and regenerative medicine are also discussed. Despite their considerable potential, current gastric organoid models remain limited: conventional epithelial organoids lack stromal, immune, neural, and vascular components; PSC-derived organoids contain only a limited mesenchymal population; and more complex engineered multicellular tissues still depend largely on in vivo engraftment for maturation. Variable maturation, dependence on poorly defined extracellular matrices, and insufficient standardization further limit their translation.

Cells doi: 10.3390/cells15211971

Authors: Nils Schellekens Jutta Horwath-Winter Sophia Bittner Wolfgang Schuh Katharina Jüngert Ingrid Zahn Martin Schicht Friedrich Paulsen

This study investigated the distribution of IgA-expressing cells in human ocular tissues and IgA subclass profiles in Schirmer strip extracts from patients with dry eye disease (DED) and primary Sjögren’s syndrome (pSS). Immunofluorescence staining was performed on paraffin-embedded lacrimal gland, conjunctival, and corneal tissues from body donors. Independently, Schirmer strip extracts from healthy controls (n = 18), patients with DED (n = 75), and patients with pSS (n = 10) were analyzed. Total protein concentrations were determined using a bicinchoninic acid assay, while total IgA, IgA1, and IgA2 were quantified by ELISA and normalized to total protein. CD38+ and CD38− IgA-expressing cells were detected in the lacrimal gland and conjunctival stroma, with IgA1-expressing cells predominating over IgA2-expressing cells. No IgA-positive cells were detected in the examined corneal sections. Total protein concentrations were lower in pSS than in DED. Normalized IgA2 abundance was reduced in both DED and pSS compared with controls, while normalized total IgA was additionally reduced in pSS. Unnormalized extract concentrations showed broadly similar patterns. These independent findings demonstrate differences in recovered IgA subclass profiles, although differences in collection materials and patient characteristics preclude attributing these alterations exclusively to the underlying diseases.

Cells doi: 10.3390/cells15211972

Authors: Jillian P. Gady Shanjida Akter Mark VandeWiele Feng Dong

Premenopausal women without diabetes have a well-established cardiovascular advantage over men of the same age, mainly due to the benefits of estrogen, which has vasoprotective, anti-inflammatory, and antioxidant effects on the vascular endothelium. However, women with type 1 or type 2 diabetes have a 40–50% higher relative risk of coronary heart disease and suffer a greater burden of heart failure and cardiovascular mortality compared with diabetic men. The mechanisms involved in the disparity are not well understood and are seldom considered in clinical guidelines or trials, where women are underrepresented. The paradoxical relationship between chronic hyperglycemia and estrogen signaling plays a critical role, since estradiol, rather than having its usual cardioprotective effect, instead leads to oxidative stress and endothelial dysfunction. This occurs through eNOS uncoupling, activation of the AGE-RAGE axis, and a shift towards pro-inflammatory signaling via estrogen receptors. Sex-related differences in body composition and the distribution of fat tissue also make a significant contribution to the variations in cardiovascular risk. Modifiable lifestyle factors, especially physical activity and skeletal muscle mass, are important targets for intervention, with evidence showing that their benefits on endothelial function, body composition, and systemic inflammation are particularly noticeable in females. In addition, the sex-specific cardiovascular responses to treatments such as glucagon-like peptide-1 receptor agonists (GLP-1RAs) and sodium-glucose cotransporter-2 (SGLT2) inhibitors should be investigated more closely. This review summarizes the current evidence, identifies critical knowledge gaps, and proposes a framework for sex-informed cardiovascular prevention in diabetes.

Cells doi: 10.3390/cells15211970

Authors: Yang Lou Qian Zhu Yuhang Tao Yameng Chen Shiquan Chen Zuwen Zhang Hongqiang Cheng Guosheng Fu Chenyang Jiang

Atrial fibrillation (AF) is the most prevalent sustained cardiac arrhythmia worldwide; hypertension is one of the most important risk factors for AF and contributes to both its initiation and maintenance. This study investigated the contribution of enolase 1 (ENO1) to hypertension-associated atrial remodeling and susceptibility to pacing-induced AF. Long-term hypertension was associated with increased AF inducibility in rats, and atrial transcriptomic analyses at two stages of hypertension identified persistent ENO1 upregulation. ENO1 was also increased in two external human left atrial AF datasets. An angiotensin II (Ang II)-induced atrial cardiomyocyte injury model was constructed to evaluate the role of ENO1 in Ang II-induced cardiomyocyte hypertrophy. In Ang II-stimulated primary neonatal rat atrial cardiomyocytes, siRNA-mediated ENO1 knockdown attenuated hypertrophic and oxidative-stress-related changes and was accompanied by lower PI3K/AKT phosphorylation and NADPH oxidase 4 (NOX4) expression. AP-III-a4 treatment produced concordant changes in selected cellular endpoints. PI3K inhibition reduced NOX4 expression and intracellular hydrogen peroxide levels. In ENO1-silenced cardiomyocytes, PI3K activation with 740Y-P partially increased both PI3K/AKT phosphorylation and NOX4 expression. In Ang II-infused hypertensive mice, AP-III-a4 treatment reduced atrial enlargement, fibrosis, PI3K/AKT phosphorylation, NOX4 expression, and atrial reactive oxygen species signals; it also attenuated atrial effective refractory period shortening. AP-III-a4 significantly shortened the duration of pacing-induced AF and was associated with numerically lower AF inducibility, although the latter difference did not reach statistical significance. Collectively, these findings support the involvement of PI3K/AKT signaling in ENO1-associated regulation of NOX4 and suggest that this relationship may contribute to the observed attenuation of hypertension-associated atrial remodeling.

Cells doi: 10.3390/cells15211969

Authors: Kyosuke Oda Arnel Belmonte David Boulifard Silvia Jakubski Lindsey White Charmagne Beckett Andrew Letizia Chad Porter Eileen Villasante Yuliya S. Johnson Wathsala Wijayalath

Extracellular vesicles (EVs) carry status quo messages across the body that can help decode the long-term impact of Post-Acute Sequelae of SARS-CoV-2 infection (PASC). Here, we analyzed paired peripheral blood mononuclear cell (PBMC)-EVs (pre- and 6–8 months post-infection) collected from a subset of young United States (US) Marines with and without a history of PASC symptoms lasting >30 days post-infection, who participated in the COVID-19 Health Action and Response for Marines (CHARM) studies. Integrated nanospray LC-MS/MS, STRING and Reactome analysis of PBMC-EVs identified distinct proteomic signatures and biological pathways associated with different PASC symptoms. An individual who reported PASC headache and drowsiness (n = 1) had elevated histones, prelamin A/C, lysozyme C and S100-A9, among other proteins enriched with cellular stress pathways, including DNA damage, oxidative/metabolic/cytoskeletal stress, senescence, neutrophil degranulation and neurodegenerative-like diseases. Conversely, some participants who reported pulmonary PASC (n = 5, pooled) had elevated histones but decreased fibrinogen, integrin β-3 and S100-A9, mapped to senescence, platelet dysfunction, and immune exhaustion pathways. Non-PASC individuals (n = 9) exhibited active inflammation resolution, without returning to the pre-infection homeostasis even after 6–8 months post-infection. These distinct PBMC-EV proteomic signatures suggest potential long-term impact of long COVID with implications for the public health and mission readiness of US service members.

Cells doi: 10.3390/cells15211968

Authors: Gabriela Zamora-Herrera Rubí Romo-Rodríguez Jebea A. López-Blanco Laura Alfaro-Hernández Diana Casique-Aguirre Juan Carlos Núñez-Enríquez Michael Schnoor Dalia Ramírez-Ramírez Rosana Pelayo

In the original publication [...]

Cells doi: 10.3390/cells15211967

Authors: Michael Linnebacher Sandra Schwarz Clemens Schafmayer

Temperature is a fundamental physical variable affecting protein folding, proteostasis, cell-cycle kinetics, and immune function, yet cancer cell biology is performed almost exclusively at a fixed 37 °C, and distinct thermal phenomena are often conflated under the single term “hyperthermia.” This review examines the biology of mild, fever-range temperature excursions of approximately 38–41 °C, which are mechanistically distinct from externally induced hyperthermia. We synthesize three strands of evidence that have rarely been considered together. First, tumor cells display heterogeneous thermal sensitivity, with differences in growth, cell-cycle responses, and thermal tolerance that may reflect intrinsic molecular features and proteostatic reserve, including the heat-shock response mediated by HSF1 and HSP70/HSP90. Second, fever-range temperatures remodel antitumor immunity across innate and adaptive compartments, affecting immune-cell trafficking, membrane organization, metabolism, and tumor–immune recognition. Third, the clinical record appears controversial: febrile infections have been associated with lower subsequent cancer risk, whereas postoperative fever has been associated with worse prognosis. We argue that these observations cannot be interpreted without considering host state, timing, and the biological context in which temperature elevation occurs. The three evidence streams are of unequal strength: cell-intrinsic thermal sensitivity and fever-range immune effects rest on direct experimental data, much of it from colorectal cancer, which we adopt as the principal proof-of-concept model, whereas the clinical associations are largely retrospective, and we treat them with correspondingly different confidence. We therefore frame a central testable question: is cell-intrinsic thermal sensitivity a cause or a correlate of fever’s effects on cancer? We propose an experimental agenda that separates cell-autonomous thermal responses from host-mediated effects and establishes temperature as an explicit, biologically informative variable in cancer research.

Cells doi: 10.3390/cells15211966

Authors: María Norte-Muñoz Joaquín Cantón-Sandoval Fernando Lucas-Ruiz Victoriano Mulero Marta Agudo-Barriuso David García-Bernal

Stem cell-based therapies have emerged as promising strategies for neurodegenerative disorders, partly supported by the concept of central nervous system immune privilege, which has encouraged xenogeneic transplantation approaches under the assumption of limited host immune responses. However, the immunological consequences of xenogeneic cell delivery into the central nervous system remain poorly characterized. In this study, we investigated the immune response elicited by intravitreal transplantation of human bone marrow-derived mesenchymal stromal cells (BM-MSCs) into healthy mouse retinas. Twenty-one days after transplantation, retinal CD45+ and/or CD11b+ cells, including microglia, macrophages, and infiltrating leukocytes, were isolated and subjected to exploratory single-cell RNA sequencing. Candidate immune cell populations and pathways identified transcriptomically were subsequently evaluated by immunostaining, flow cytometry, and plasma antibody analyses. Xenogeneic BM-MSC transplantation induced marked transcriptional changes consistent with immune activation, enhanced antigen presentation, and leukocyte recruitment, features that are not typically associated with central nervous system homeostasis. These findings were supported by complementary cellular and systemic analyses, collectively revealing innate and adaptive immune responses directed against the transplanted human cells. The results demonstrate that xenogeneic BM-MSC transplantation can overcome local immune regulatory mechanisms and trigger robust immune activation within the healthy retina. These findings identify previously underappreciated immunological barriers to xenogeneic stem cell-based therapies and reveal coordinated innate and adaptive immune responses despite the presumed immune privilege of the CNS. Consideration of these immune responses will be essential for the safe and effective translation of regenerative therapies.

Cells doi: 10.3390/cells15211965

Authors: Yuanyuan Pan Lumiao Zhang Rongyuan Zhu Fangfang Sun Guofeng Wu

Background: Bone marrow mesenchymal stromal cells (BMSCs) are widely used as seed cells in bone tissue engineering. However, sufficient cell quantities for effective engineering typically require prolonged in vitro expansion of BMSCs, which can lead to cellular senescence after excessive subculturing. Objective: This study aimed to investigate the effects of human induced pluripotent stem cell (hiPSC) extract on replicative senescence, clonogenic ability, proliferation-associated characteristics, and osteogenic differentiation capacity of human alveolar bone-derived mesenchymal stromal cells (AB-MSCs). Methods: AB-MSCs were isolated from alveolar bone fragments discarded during dental implant surgery. Cells were divided into three experimental groups: P3 AB-MSCs (Young), P10 AB-MSCs (senescence-associated, YRS), and P10 AB-MSCs treated with hiPSC extract (YRSiE). Senescence markers, clonogenic ability, proliferation-associated characteristics, osteogenic potential, mRNA sequencing and pathway-related expression changes were compared among groups. Small RNA sequencing on hiPSCs and AB-MSCs with qRT-PCR validation were performed to identify candidate miRNAs in hiPSC extract that were differentially expressed in this setting. Results: In the presence of hiPSC extract, senescent AB-MSCs showed altered morphology, a lower proportion of β-galactosidase-positive cells, higher Ki67 expression, and lower expression of the senescence-associated genes and proteins P16 and P21. Treated senescent AB-MSCs also showed higher proliferation-associated metabolic activity, clonogenicity, and osteogenic differentiation than untreated senescent cells. Pluripotency-related genes (NANOG, OCT-4) and osteogenic markers (ALP, RUNX2, OCN) were higher in the treated group. qRT-PCR validation of small RNA sequencing data showed that miR-514a-3p, miR-582-5p and miR-520c-3p were upregulated in senescent AB-MSCs after hiPSC extract treatment. Concurrently, pathway enrichment and pathway-related expression changes were consistent with an association with the NF-κB pathway. Conclusion: In this in vitro study, hiPSC extract attenuates senescence-associated phenotypes in AB-MSCs, and the observed transcriptional and pathway-related expression changes were consistent with NF-κB pathway enrichment, with the identified miRNAs upregulated in treated cells and previously reported to target NF-κB pathway components. However, these mechanisms require further validation.

Cells doi: 10.3390/cells15211964

Authors: Vassa D. Davydova Anastasia A. Zotova Nikolay M. Kapranov Dmitry V. Dianov Yulia O. Davydova Tatiana A. Nenasheva Irina Y. Zaichenko Sergei D. Losev Kseniia A. Nikiforova Irina V. Galtseva Apollinariya V. Bogolyubova

The development of chimeric antigen receptor (CAR) T cell therapies for acute myeloid leukemia (AML) remains challenging, underscoring the need for reliable preclinical models. Although AML cell lines are widely used in CAR-T studies, their ability to reflect clinically relevant antigen expression patterns and functional characteristics of primary AML cells has not been systematically evaluated. We characterized nine AML cell lines (KG-1, HL-60, Kasumi-1, OCI-AML2, OCI-AML5, THP-1, U-937, MV4-11, and MOLM-13) as targets for in vitro CAR-T studies. We assessed the expression of established CAR-T target antigens (e.g., CLL1, CD33), cell lines similarity to minimal residual disease (MRD) cells and matched healthy monocytes from 161 AML patients, the expression of emerging AML-associated antigens (Siglec-6, CSF1R, and CD70), growth behavior, and susceptibility to non-specific killing by non-transduced T cells. AML cell lines demonstrated substantial heterogeneity in antigen expression with several cell lines showing antigen densities exceeding those observed in AML blasts. Several widely used lines, including U-937, were susceptible to non-transduced T-cell cytotoxicity, potentially confounding CAR-T functional assays and overestimating CAR-T activity in vitro. The examined parameters were integrated into a scoring framework to rank AML cell lines according to their suitability for in vitro CAR-T studies. Based on this framework and additional assay-related considerations, OCI-AML2, OCI-AML5, MV4-11, and MOLM-13 were identified as most suitable for in vitro evaluation of CAR-T products, particularly those targeting CD33 and CLL1. These findings may facilitate more informed selection of AML cell lines for preclinical development of CAR-T therapies for AML.

Cells doi: 10.3390/cells15211963

Authors: Jianhong Dong Xianji Wei Yifan Qian Yiming Qian Wei Zhang

Bone remodeling requires coordinated transcriptional regulation across osteoblasts, osteoclasts, and osteocytes. Histone acetylation connects cellular metabolism to these lineage-specific programs, yet its regulatory effects depend not simply on total cellular acetyl-CoA abundance, but also on metabolic carbon routing, subcellular acetyl-CoA availability, and the targeting of chromatin-modifying enzymes to specific genomic regions. This review examines how these processes shape skeletal-cell differentiation, function, and communication within the bone-remodeling unit. We integrate evidence linking acetyl-CoA metabolism to histone acetylation across skeletal lineages, highlighting the distinct contributions of metabolic substrate utilization, acetylation and deacetylation machinery, and locus-specific chromatin regulation. We further discuss how mechanical loading, aging, inflammation, and metabolic disease reshape these regulatory networks and influence the balance between bone formation and resorption. Particular attention is given to the distinction between metabolic changes and their chromatin consequences, as well as to the experimental approaches needed to resolve acetyl-CoA compartmentalization and trace metabolic carbon into histone acetyl groups. Finally, we evaluate therapeutic opportunities and challenges in targeting metabolic–chromatin pathways while preserving physiological bone remodeling. Integrating metabolic routing, chromatin regulation, and intercellular communication provides a framework for understanding skeletal adaptation and identifying new avenues for therapeutic development.

Cells doi: 10.3390/cells15211960

Authors: Martina Mietto Maria Sofia Falzarano Annarita Armaroli Fernanda Fortunato Stefania Bigoni Roberta Schininà Beatrice Vianello Federico Grasso Daniela Benati Andrea Cerase Milena Cau Maria Antonietta Melis Xiao Dang Mingyan Fang Alessandra Recchia Salvatore Fioriniello Floriana Della Ragione Alessandra Ferlini

NF-κB-activating protein (NKAP) gene maps to Xq24 and encodes a nuclear protein involved in NF-κB transcription factor activation. Missense variants in NKAP have previously been reported in males with X-linked intellectual disability. Here, we identified a recurrent, novel frameshift variant, c.172del (p.His58Ilefs74*), by targeted X-chromosome sequencing (X-TRS) in seven females from a Sardinian family showing fully skewed X-chromosome inactivation (XCI). To investigate the functional effect of the mutation, we knocked out (KO) and knocked down (KD) NKAP in healthy donor fibroblasts and mouse embryonic stem cells, respectively. We performed RNA sequencing, immunofluorescence, and Western blot analysis on fibroblasts derived from the index subject. We demonstrated that the NKAP frameshift variant leads to monoallelic wild-type (WT) transcription and does not directly affect XCI, indicating secondary rather than primary XCI skewing. NKAP-edited human fibroblasts underwent cell death, leading to the positive selection of WT cells. The NKAP correctly localized to the nucleus but showed reduced expression. We also showed that XIST expression was not affected by this NKAP mutation, further excluding a primary XCI event. Our findings demonstrate that this novel NKAP frameshift variant causes early cell death and post-mitotic negative selection of mutant cells, resulting in a fully secondary XCI. Female carriers are healthy and fertile, whereas affected males are likely nonviable. In conclusion, our results provide insights into both the role of NKAP in cells and novel clinical implications for human fertility.

Cells doi: 10.3390/cells15211961

Authors: Florence Fioretti Guoqiang Hua Nadia Benkirane-Jessel Rana Smaida

For young and middle-aged patients with irreparable meniscal tears, preserving or restoring meniscal tissue is critical to maintaining joint function and delaying osteoarthritis. Yet, when repair is not feasible, partial meniscectomy remains common and can compromise the load-bearing function of the knee, underscoring a highly unmet medical need for regenerative strategies that restore functional tissue rather than simply relieve symptoms. Induced pluripotent stem cells (iPSCs) offer a scalable and potentially patient-matched source of progenitor cells for meniscal regeneration, with iPSC-derived cartilage already entering clinical study for articular cartilage defects. However, translating this progress to the meniscus presents a fundamental biological challenge: restoring a specialized, zonally organized tissue rather than simply generating sufficient cells. The meniscus is a structurally, mechanically, and biologically graded tissue with distinct inner, transitional, and outer zones. Consequently, generating fibrocartilage is unlikely to reproduce native meniscal function. In this review, we examine iPSC-based strategies through the framework of zone-specific meniscal regeneration, focusing on meniscal fibrochondrocyte generation and reconstruction of native tissue architecture. We compare biofabrication strategies against native structural, vascular, and mechanical benchmarks, and outline the preclinical, manufacturing and regulatory requirements for translating iPSC-derived meniscal constructs toward patient-centered clinical application.

Cells doi: 10.3390/cells15211962

Authors: Sunjiangnan Wu Linggeng Kong Miao Zhu Bing Yao Shanmeizi Zhao Li Chen

The stress-inducible molecular chaperone HSP90α, encoded by Hsp90aa1, plays an essential role in male reproduction. Systemic Hsp90aa1 deficiency causes pachytene arrest, germ cell loss, and azoospermia. However, systemic models cannot distinguish effects originating in germ cells from those mediated by the testicular somatic microenvironment. Sertoli cells sustain germ cell development through physical support, nutrient supply and regulatory signaling. The function of HSP90α in Sertoli cells has previously been studied mainly in cultured cells, with a lack of in vivo evidence. To investigate its role in Sertoli cells, we generated Hsp90aa1flox/flox; Amhr2-Cre+ conditional knockout (CKO) mice. HSP90α was markedly reduced in Sertoli cell-enriched fractions from CKO testes, and testicular immunofluorescence showed reduced signal in Sertoli cell regions. Despite this reduction, CKO males remained fertile and showed no significant abnormalities in testicular morphology, spermatogenic progression, the measured sperm parameters, Sertoli cell counts, or the localization of the junction-associated proteins. However, total germ cell counts were modestly reduced in CKO testes, with the reduction primarily observed in spermatocytes. These findings indicate that Sertoli-cell Hsp90aa1 deletion is associated with a modest reduction in spermatocyte numbers but does not overtly impair male fertility under the conditions examined. The contrast with systemic Hsp90aa1 deficiency suggests that the requirement for HSP90α in male reproduction is cell type-specific.

Cells doi: 10.3390/cells15211959

Authors: Nikolas Ala-Kokko Emory Gregory Alan Woessner Bing Guan Winifred Middleton Donald J. Johann Narasimhan Rajaram Timothy J. Muldoon Younghye Song

Metabolic reprogramming is a well-established hallmark of cancer, contributing to adverse behaviors like invasion, metastasis and recruitment of tissue-resident cells to the tumor microenvironment. Among recruitment capabilities, innervation has emerged as an active contributor of disease progression. Despite association with poor patient prognosis, mechanisms governing nerve recruitment at various stages of progression remain poorly understood. This study investigates how different stages of breast cancer cells are associated with the metabolic link of cancer and their pro-axonogenic capability. Here, we used human isogenic MCF10 mammary epithelial cell line series, three-dimensional collagen hydrogels, human adipose stromal cells (ASCs) and freshly isolated rat sensory neurons. Multiphoton optical redox imaging coupled with ATP production rate assays validated direct metabolic inhibition and highlighted the utilization of glutamine in progressed cells. We identified the expressions of brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) from pre-malignant and malignant cultures, respectively. Interestingly, when treated with metabolic inhibitors, monocultures of MCF10 cells reduced neurotrophin release while cocultures with ASCs showed no or even elevated neurotrophin secretion. Together, these findings raise the possibility that breast tumor innervation is shaped in part by stage-specific metabolic-neurotrophic coupling, presenting a concrete mechanistic question for future investigation.

Cells doi: 10.3390/cells15211958

Authors: Bianca Fiorillo Alessandra Guarracino Oriana Vitale Federica Moraca Bruno Catalanotti

The rapid increase in experimentally resolved G protein-coupled receptor (GPCR)–biologic complexes has substantially improved our understanding of the molecular basis of receptor recognition and modulation. However, despite the growing number of available structures, a comparative framework integrating the major GPCR classes is still lacking. In this review, we analysed experimentally resolved GPCR–biologic complexes across receptor Classes A, B, C, and F to identify the structural principles governing biologic recognition. Our analysis shows that the architecture of each GPCR class shapes distinct recognition paradigms, while two interdependent structural variables emerge across the superfamily: the accessibility and organization of receptor surfaces and the conformational states associated with these regions. Class A receptors primarily expose compact extracellular surfaces, Class B receptors exploit large extracellular domains that support modular ligand recognition, whereas Class C and Class F receptors rely on structured extracellular domains (Venus Flytrap and cysteine-rich domains, respectively) that enable conformational selection and allosteric regulation. Together, these observations establish a structural framework linking receptor accessibility and conformational dynamics to biologic recognition and pharmacological modulation. This framework provides a conceptual foundation for the rational design of next-generation GPCR-targeted biologics with improved selectivity, efficacy, and functional precision.

Cells doi: 10.3390/cells15211957

Authors: Mykola Zdioruk Eleni Panagioti Noe B. Mercado Vincent van Unen Ennio Antonio Chiocca Michal O. Nowicki James A. Lederer Sean E. Lawler Charles H. Cook

Glioblastoma (GBM) is a highly aggressive brain tumor with no effective treatments. It is characterized by rapid progression and marked heterogeneity, which contributes to therapeutic resistance. Human cytomegalovirus (hCMV), a highly prevalent β-herpesvirus, has been associated with GBM in numerous studies, and both antiviral drugs and hCMV-targeted immunotherapies have shown evidence of therapeutic activity in clinical trials. However, the mechanisms underlying this association remain poorly understood. To address this, we previously developed a murine model of GBM with murine CMV (mCMV) infection, which revealed accelerated tumor progression in infected mice. Here, we investigated whether tumor heterogeneity influences the effects of mCMV infection on intracranial tumor growth across four murine GBM models (GL261, CT-2A, M005 and Mut3). Tumor progression was monitored longitudinally by MRI and evaluated by survival analysis, histopathology and characterization of the tumor microenvironment. Systemic mCMV infection was associated with accelerated GBM progression across all models, characterized by increased tumor growth and significantly reduced survival. Enhanced tumor progression was accompanied by increased vascularization, elevated myeloid cell activation and intratumoral viral activity in a subset of tumors. Mass cytometry (CyTOF) revealed remodeling of the tumor immune landscape, including a marked increase in intratumoral virus-specific CD8+ T cells, highlighting a potential avenue for therapeutic targeting. Together, these findings indicate that CMV is associated with accelerated GBM progression across genetically distinct murine models, accompanied by remodeling of the tumor microenvironment, including increased vascularization and altered immune infiltration.

Cells doi: 10.3390/cells15211956

Authors: Albert Despot Ivan Sabol Zlatko Hrgović Ana-Marija Despot Ivica Vlatkovic Mara Tešanović Dora Fureš Sanja Malinac Malojčić Vesna Ćosić Nikola Sinković Rajko Fureš

Cervical cancer and breast cancer represent two serious challenges in modern gynecological oncology. Despite their anatomical accessibility for screening, significant investments in research, and the development of therapeutic strategies, their global incidence and mortality rates continue to rise. Interstitial spaces, paracrine signaling, hormonal imbalance linked to xenoestrogens (XE) and the consequent inhibition of apoptosis constitute a common, yet unfortunately insufficiently recognized, link in the carcinogenesis of both cervical and breast cancer. The aim of this paper is to supplement the classical paradigm of cervical and breast carcinogenesis by incorporating a broader context currently missing from the understanding of both forms of cancer. XEs are widely distributed chemical substances, found in a wide range of everyday products, food and environment, that mimic the action of endogenous estrogen. In the world of basic science, the connection between estrogen dominance and apoptosis avoidance is unquestionable and can undeniably impact progression of breast and cervical cancer. In the development of breast cancer, the association between XE facilitated estrogen dominance and arrested apoptosis is of great importance. In the pathogenesis of cervical cancer, the activity of viral oncogenes E6/E7 is the main driver of the pathological condition; however, in the context of arrested apoptosis, there is an area of overlap between estrogen dominance and HPV virus action. In this sense, estrogen dominance, triggered by the agonistic action of XEs on the membrane-bound GPER receptor of fibroblasts, is a cofactor in cervical carcinogenesis that remains insufficiently researched.

Cells doi: 10.3390/cells15211955

Authors: Giuseppe Zardo Gloria Pasqualini Gianfranco Catalano Alessandra Zaza Nelida Ines Noguera

Metabolic reprogramming is increasingly recognized as a fundamental component of acute myeloid leukemia (AML) biology, but AML cannot be defined by a single metabolic phenotype. Leukemic cells occupy dynamic metabolic states shaped by differentiation, genetic background, the bone marrow microenvironment, and therapeutic pressure. In this Review, we propose that metabolic biomarkers should capture functional dependencies and metabolic plasticity rather than static metabolic states, with functional dependencies defined as the reliance of AML cells on a specific metabolic pathway or process for their survival, proliferation or maintenance, rather than a simple glycolytic-versus-oxidative dichotomy. Normal hematopoietic stem cells (HSCs), leukemic stem cells (LSCs), and bulk AML blasts display distinct metabolic programs. Quiescent HSCs maintain a low-energy state, whereas LSCs rely strongly on mitochondrial oxidative phosphorylation (OXPHOS), amino acid metabolism, mitochondrial quality control, and antioxidant defenses. AML blasts exhibit greater metabolic heterogeneity and can switch between glycolytic and oxidative pathways. Importantly, increased mitochondrial activity does not necessarily indicate an LSC phenotype, as therapy-resistant AML cells may acquire enhanced OXPHOS, fatty acid oxidation (FAO), and mitochondrial fitness as adaptive states. Acute promyelocytic leukemia (APL) provides a model of lineage-specific metabolic rewiring, with PML::RARA suppressing glycolysis while promoting FAO, TCA cycle activity, OXPHOS, and arginine-associated anaplerosis. We propose that metabolic biomarkers should identify functional dependencies and escape routes. Integrating metabolic profiling with multi-omics approaches may enable identification of AML subgroups with distinct vulnerabilities and guide rational combination therapies.

Cells doi: 10.3390/cells15211954

Authors: Martina Milella Francesco Lasorsa Monica Rutigliano Alessandro Caniglia Emanuele Naglieri Savio Domenico Pandolfo Pasquale Ditonno Giuseppe Lucarelli

Renal cell carcinoma (RCC), particularly its clear-cell subtype (ccRCC), is characterized by metabolic reprogramming and a complex tumor microenvironment that influence responses to immune checkpoint inhibitors (ICIs). Despite clinical advances, therapeutic resistance remains a major hurdle. Here we review the molecular determinants governing ccRCC sensitivity and resistance to ICIs, integrating insights from genomic alterations such as PBRM1 and BAP1 mutations, immune cell exhaustion dynamics, stromal and metabolic barriers, and the role of endogenous retroviral antigens. We discuss how these factors modulate immune evasion and impact therapeutic outcomes. Emerging strategies including biomarker-driven patient stratification, HIF-2α inhibition, and personalized immunotherapies show promise in overcoming resistance. Our synthesis highlights the necessity of multi-omics approaches to tailor immunotherapy, aiming to enhance durable responses and survival in ccRCC patients.

Cells doi: 10.3390/cells15211953

Authors: Mariangela Centrone Ines Angelini Mariagrazia D’Agostino Annarita Di Mise Barbara Barile Grazia Paola Nicchia Giampaolo Morciano Stine Julie Tingskov Xabier Sørtvedt Birgitte Mønster Christensen Marianna Ranieri Giovanna Valenti Rikke Nørregaard Grazia Tamma

Lithium is widely used to treat psychiatric disorders, including bipolar disorder. However, long-term lithium therapy is frequently associated with acquired nephrogenic diabetes insipidus (NDI), a disorder characterized by impaired urinary concentrating ability and excessive water loss. Tamoxifen (TAM), a selective estrogen receptor modulator, has previously been shown to alleviate lithium-induced NDI by restoring AQP2 and AQP3 expression. Here, we investigated whether necroptosis-related signaling, primary cilium remodeling, and epithelial–mesenchymal transition (EMT)-like changes contribute to lithium-induced collecting duct alterations and whether TAM modulates these responses. Using complementary in vitro, ex vivo, and in vivo approaches, we found that lithium significantly reduced the expression of the necroptosis-related proteins receptor-interacting protein kinase 3 (RIPK3) and mixed lineage kinase domain-like protein (MLKL), whereas TAM prevented their downregulation. Lithium also induced primary cilium elongation and promoted EMT-like changes, characterized by reduced E-cadherin and increased vimentin expression, which were attenuated by TAM. Collectively, these findings demonstrate that lithium profoundly alters collecting duct epithelial homeostasis by modulating the abundance of necroptosis-related proteins, primary cilium morphology, and epithelial phenotype. The ability of TAM to counteract these alterations indicates that its protective effects extend beyond aquaporin regulation, supporting its potential as a therapeutic strategy for lithium-induced NDI.

Cells doi: 10.3390/cells15211952

Authors: Irene Fancello Veronica Candino Daniel Gamo Gonzalez Maurizio Pesce Gloria Garoffolo

Non-coding RNAs (ncRNAs) have emerged as major regulators of gene expression and cellular homeostasis, with growing evidence supporting their involvement in cardiovascular physiology and disease. This review summarizes the classification, mechanisms of action, and functional roles of ncRNAs, with particular focus on mechano-ncRNAs involved in cardiovascular remodeling and fibrosis. Different classes of ncRNAs are discussed in relation to their contributions to vascular, valvular, and myocardial homeostasis, as well as to pathological processes including inflammation, endothelial dysfunction, and fibrotic responses. Special emphasis is placed on biomechanical stimuli, including increased matrix stiffness, cyclic strain, and altered shear stress, which can modulate ncRNA expression and activity through mechanotransduction pathways. Notable examples of mechano-ncRNAs include miR-21, miR-29, miR-132, MIAT, MALAT1, and MEG3, which have been implicated in maladaptive cardiac remodeling and fibrosis. Concurrently, we highlight the importance of technical advances in ncRNA discovery and cardiac-specific delivery systems, including viral and nanoparticle-based vectors, for the development of ncRNA-based therapeutic strategies with potential for clinical translation. Overall, ncRNAs are shown to represent promising biomarkers and therapeutic targets in cardiovascular diseases, particularly in mechanically driven pathological conditions. However, challenges related to delivery specificity, stability, off-target effects, and safety remain and will need to be addressed to enable the clinical translation of ncRNA-based therapies in precision cardiovascular medicine.

Cells doi: 10.3390/cells15211951

Authors: Ling Wei Michael Qize Jiang Yingying Zhao Daiana Takashima Yi Li Khan Hekmatyar Lingyue Wu Ananth Sastry Jenny J. Yang Shan Ping Yu

Sensory dysfunction, such as olfactory and visual deficits, is a notorious symptom during the progression of Alzheimer’s disease (AD) and related dementia (ADRD). The causal factor/s and structural disruptions underlying the sensory dysfunction in the AD/ADRD brain are not well understood. The NMDA receptor subunit GluN3A (NR3A) negatively regulates the receptor activity to uphold cellular Ca2+ homeostasis. In GluN3A knockout (KO) mice, animals spontaneously develop chronic neurodegenerative excitotoxicity, cognitive decline, and AD-like pathology. Using diffusion tensor imaging (DTI), we examined neuronal network destruction together with sensory dysfunction in the GluN3A KO mouse. Specific attention was given to the selective vulnerability of the olfactory and visual nuclei, white matter injury in regional connections between subcortical centers and the cortex and hippocampus. Compared to age-matched wild-type (WT) controls (male), consistent with our early reports in younger ages, 18-month-old aging/aged GluN3A KO mice (male) suffered significant olfactory, visual and social deficits, accompanied by marked Aβ42 deposition in the brain. Ex vivo MR images of the GluN3A KO and control brains were acquired on the 7T MRI system, using a four-channel mouse array RF coil. High-resolution 3D T2-weighted RARE anatomical and DTI scans were performed. The DTI analysis revealed that GluN3A KO mice developed brain atrophy, involving significant damage in the white and gray matter of the olfactory and visual systems and impaired pathways among the superior colliculus, lateral geniculate nucleus, and the hippocampus and/or the cortex. This investigation disclosed a receptor-mediated neurodegeneration that explains sensory syndromes featured in a sporadic AD mouse model.

Cells doi: 10.3390/cells15211950

Authors: Taketo Taguchi Pamela Maher

Aging is accompanied by a progressive disruption of brain homeostasis, yet the molecular mechanisms underlying this age-associated deterioration remain poorly understood. Microglia play a critical role in maintaining brain homeostasis by clearing cellular debris and neurotoxic protein aggregates. Here, we investigated the effects of methylglyoxal (MG), a highly reactive endogenous metabolite that accumulates with aging, on microglial functions using the murine microglial cell line BV2. We assessed the effects of MG exposure on multiple aspects of microglial biology in both control and bacterial lipopolysaccharide (LPS)-treated cells, including phagocytic activity, cellular proliferation, cytokine secretion, mitochondrial bioenergetics, mitochondrial membrane potential, and intracellular and mitochondrial reactive oxygen species. We found that MG significantly impaired mitochondrial bioenergetics, leading to reduced cellular energy production, and markedly suppressed multiple essential microglial functions, including phagocytosis. Given the high reactivity of MG with arginine, we also investigated whether arginine depletion contributes to these functional impairments. Arginine depletion recapitulated the impairment of mitochondrial bioenergetics and microglial function induced by MG, demonstrating that arginine availability is a critical determinant of microglial metabolic fitness and immune function. These findings suggest that the age-associated accumulation of MG impairs mitochondrial bioenergetics and microglial functions, thereby potentially accelerating neurodegenerative processes.

Cells doi: 10.3390/cells15211949

Authors: Sheng Chen Yufan Luo Qiuzhi Zhou Hong Chen

Metabolic abnormalities in rheumatoid arthritis (RA) differ across cell populations and anatomical compartments. Glycolytic synovial fibroblasts coexist with blood T-cell populations showing impaired glycolytic induction or mitochondrial function, while macrophage metabolism can sustain inflammation or tissue protection. These differences complicate systemic metabolic inhibition. This narrative review integrates human RA studies, experimental arthritis and mechanistic work in other systems to identify dependencies that support defined pathogenic functions. We examine metabolite transport, nutrient adaptation and nutrient and energy sensing, distinguishing causal perturbation from descriptive signatures and preclinical hypotheses from clinical efficacy signals. Systemic cardiometabolic disease and nutritional interventions provide a clinical context for interpreting local metabolic findings. Small trials of repurposed drugs suggest possible benefit during treatment, but do not establish durable metabolic correction. We propose that a claim of reprogramming requires persistent functional change after drug removal, exclusion of residual exposure and selective cell loss, and preservation of immune regulation and host defence. These criteria could guide more selective metabolic interventions and their evaluation in patient tissue.

Cells doi: 10.3390/cells15211948

Authors: Hao Yang Yunqian Zeng Yiyang Teng Qi Zhao Minbo Hou Zhikai Cheng Xuemei Zhang Xijie Wang

Understanding early cardiac developmental abnormalities requires linking tissue phenotypes to the cellular populations produced during differentiation. We established a cardiac organoid platform derived from human induced pluripotent stem cells (hiPSCs) and combined phenotypic assays, time-course and day-9 single-cell transcriptomics, and cleavage under targets and tagmentation (CUT&Tag) to investigate developmental perturbations. Organoids contained multiple cell populations and exhibited stage-dependent cardiac transcriptional programs. Thalidomide exposure altered morphology and reduced TNNT2-positive tissue area, with lower calcium transient amplitude at 0.01 μg/mL. RNA sequencing at 10 μg/mL identified downregulation of heart and neural crest derivatives expressed 1 (HAND1), prompting a separate genetic investigation. Across three paired differentiation experiments using one characterized HAND1-knockout clone, reduced cardiomyocyte representation accompanied relative enrichment of mesodermal/mesenchymal-like cells. Selected cardiac-associated expression abnormalities persisted at day 15. Expression and chromatin occupancy analyses identified candidate regulatory associations, with locus-specific quantitative PCR supporting HAND1 occupancy at selected cardiac-associated regions. Independent genetic and functional validation is needed to test causal relationships. Together, these findings place HAND1 loss-associated tissue abnormalities in their cellular context and identify candidate occupied regions for investigating early cardiac differentiation in a human three-dimensional model.

Cells doi: 10.3390/cells15211947

Authors: Argyro Lamprou Cristina Porcheri Thimios A. Mitsiadis

Rodent incisors exhibit pronounced labial–lingual asymmetry, with enamel deposited exclusively on the labial surface and cementum on the lingual surface. This asymmetry arises early in development as the initially symmetrical dental epithelium rotates and reorients along the jaw axis. Here, we investigated the cellular and molecular mechanisms underlying this process. Quantitative cell-cycle analyses and transcriptomic profiling before and during epithelial rotation revealed accelerated proliferation, shorter cell-cycle duration, and enrichment of DNA replication and mitotic genes within the labial epithelium compared with the lingual epithelium. The evolutionarily conserved Notch signalling pathway, a regulator of cell proliferation and fate, was asymmetrically active during this period, with Notch2 expression restricted to the labial domain. Lineage tracing demonstrated that Notch2-expressing cells and their progeny preferentially contribute to the enamel-producing compartment. Epithelial-specific deletion of Notch2 produced no obvious early morphological phenotype and induced upregulation of other Notch pathway components. By contrast, pharmacological Notch inhibition reduced epithelial asymmetry and altered S-phase progression, and decreased Ccnd1 and Cdk6 expression. Together, these findings implicate Notch signalling in regional cell-cycle dynamics during early mouse incisor asymmetry, while suggesting that other Notch components may compensate for Notch2 loss.

Cells doi: 10.3390/cells15211946

Authors: Sean Michael Carr Tharmarajan Ramprasath Ming-Hui Zou

Objective: Chronic kidney disease (CKD) worsens outcomes of peripheral artery diseases (PADs), partly through impaired vascular regeneration and microvascular remodeling. CKD promotes the accumulation of uremic toxins, but their contributions to microvascular dysfunction remain poorly understood. This study investigated the effects of uremic toxin indoxyl sulfate (IS) on vascular smooth muscle cell (VSMC) function and microvascular remodeling in a mouse model of CKD. Methods: C57BL6/J mice were fed 0.25% adenine diet for 10 days to induce CKD. CKD mice underwent hindlimb ligation surgery. VSMC migration assays, microscopy, and Western blotting were utilized to elucidate the underlying mechanisms underlying the IS-induced impairment of vascular remodeling. Results: CKD mice exhibited significantly impaired blood flow restoration (44.7% relative blood flow) compared with controls (82.6% relative blood flow) accompanied by a reduced α-smooth muscle expression and decreased formation of smooth muscle-covered micro vessels in ischemic limbs. Wound healing and transwell migrations assays demonstrated IS significantly reduced VSMC motility. IS-exposure also disrupted actin cytoskeletal organization, impaired cell attachment, reduced cell size, and decreased focal adhesion formation. Mechanistically, IS reduced focal adhesion kinase (FAK) expression, impairing VSMC adhesion and migration. Reduced FAK signaling was associated with decreased N-WASP expression and suppression of the Arp2/3 complex, which regulates actin polymerization and cytoskeletal remodeling. These alterations suggest that IS disrupts the cellular processes required for VSMC recruitment and structural remodeling of the microvascular, potentially limiting vessel maturation and stabilization during ischemic recovery. Conclusions: IS impairs VSMC migration and cytoskeletal organization through the disruption of the FAK-N-WASP-Arp2/3 signaling axis, contributing to defective microvascular remodeling and impaired perfusion recovery in CKD-associated PAD. These findings identify uremic toxin-mediated cytoskeletal dysfunction as a potential therapeutic target for improving microvascular regeneration and vascular recovery in patients with CKD.

Cells doi: 10.3390/cells15211945

Authors: Diana Bellavia Sabrina Zema Maria Valeria Giuli Giulia Ferrera Samantha Cialfi Saula Checquolo Rocco Palermo Claudio Talora

In the concept of cancer evolution is implicit a causal necessity that dictates the tumor’s evolutionary trajectory. It is recognized as a dynamic process in which tumor cells continuously adapt to environmental, metabolic, and therapeutic pressures. Such adaptability relies on the progressive engagement of molecular mechanisms that under physiological conditions govern cellular flexibility and fate transitions. During tumorigenesis these regulatory systems are frequently repurposed and integrated into novel functional networks that extend far beyond their canonical roles. From this perspective, the complexity of cancer evolution does not reside merely in the accumulation of genetic alterations, but in the ability of tumor cells to preserve the plasticity necessary to continuously redefine their biological identity. The ability to switch between distinct phenotypic states and respond to selective pressures represents a fundamental hallmark of cancer progression. Among the signaling pathways implicated in these adaptive processes, Notch signaling might play a particularly prominent role. Traditionally regarded as a master regulator of cell fate specification during development, Notch signaling is increasingly emerging as a broader regulator of cellular adaptability. Rather than functioning exclusively as a developmental determinant or alternatively as a classical oncogene or tumor suppressor, accumulating evidence suggests that Notch acts as a context-dependent adaptive network capable of integrating signals generated by genomic instability, DNA damage, metabolic stress, therapeutic exposure, and microenvironmental interactions. In this review, we propose a conceptual framework in which Notch signaling is interpreted as a molecular system governing tumor adaptability. We discuss how post-translational regulators, including PLK1, PIN1, and ubiquitin-dependent mechanisms, shape Notch activity during cellular stress responses. Here, we discuss how tumor cells dynamically regulate Notch signaling and explore how context-dependent Notch signaling contributes to adaptive cellular programs thereby providing a signaling platform through which tumor cells preserve the phenotypic flexibility that underlies adaptation and evolution.

Cells doi: 10.3390/cells15211944

Authors: Hon Shing Ong Deepika Malhotra Dawn J. H. Neo Xiao Yu Ng Isha Chaudhari Gary S. L. Peh Jodhbir S. Mehta

Purpose: This study seeks to evaluate the effects of pre-treating donor corneal tissue with a Rho-associated kinase inhibitor (ROCKi), Netarsudil (AR-13324), on the viability of corneal endothelial cells (CECs) during Descemet’s membrane endothelial keratoplasty (DMEK) graft preparation and insertion using endothelium-in and endothelium-out techniques. Methods: This ex vivo comparative study utilized 24 pairs of research-grade human donor corneas preserved in Optisol-GS. One cornea from each pair was incubated in 1 µM AR-13324 for 2 h, while the contralateral cornea served as a control. DMEK tissues were prepared and simulated insertion was performed using either the endothelium-in (Endoglide) or endothelium-out (Geuder glass injector) technique. The surgeon performing the tissue preparation and insertion was masked to the ROCKi treatment allocation. Post-manipulation, tissues underwent live/dead cytotoxicity assay using Calcein-AM and Ethidium homodimer-1 staining. Percentages of live, fallout, and dead CECs were quantified via fluorescence microscopy and ImageJ (version 1.54p) analysis. Correlations between donor age, preservation duration, and cell viability were also analyzed. Results: For the endothelium-out model, live cell percentage was lower in controls compared to ROCKi-treated corneas (89.6 ± 5.35% vs. 95.1 ± 3.74%, p = 0.0001), while the percentage of dead cells was higher in controls (7.13 ± 3.71% vs. 2.16 ± 1.39%, p = 0.0017). For the endothelium-in model, live cell percentage was lower in controls compared to ROCKi-treated corneas (86.8 ± 4.81% vs. 94.4 ± 3.26%, p = 0.0001), with corresponding higher percentage fallout (8.21 ± 5.02% vs. 4.05 ± 2.82%; p = 0.0036) and higher percentage dead cells (5.04 ± 3.52% to 1.56 ± 1.13%; p = 0.0070). No significant correlations were found between donor age or preservation time and cell viability outcomes. Comparative analysis showed no significant difference in cell survivability between the two DMEK techniques with ROCKi treatment. Conclusions: Pre-incubation with the ROCKi AR-13324 significantly enhances corneal endothelial cell viability and reduces cell death during DMEK graft preparation and manipulation, regardless of the graft insertion technique used. Incorporating ROCKi treatment into tissue preparation protocols may enhance endothelial preservation, potentially expanding its clinical utility in eye banking and improving corneal graft survival.

Cells doi: 10.3390/cells15211943

Authors: Jingjing Li Chuntao Su Xuliang Luo Zhen Zhang Yi Xu Yanping Feng

Semen quality in roosters is a critical reproductive trait in poultry, yet its genetic regulatory mechanisms remain poorly understood. Sertoli cells (SCs) are essential for establishing the blood–testis barrier and supporting spermatogenesis. Our previous single-cell transcriptomic analysis identified FAM65B (Family with sequence similarity 65, member B) as a SC-specifically enriched gene that is significantly upregulated in testes of roosters with high semen quality. Given its known roles in cell proliferation, adhesion, and cytoskeletal remodeling, we systematically investigated the spatiotemporal expression pattern of FAM65B in chicken testes and its functional roles in SC physiology using a combination of in vitro and tissue-based approaches. Our results demonstrate that FAM65B is predominantly localized in the SC cytoplasm. Transcriptomic analysis revealed that FAM65B knockdown downregulated genes involved in cell adhesion, cytokine signaling, and the PI3K-Akt and NF-κB pathways. Knockdown of FAM65B significantly inhibited SC proliferation, reduced the expression of junction-associated molecules, and impaired cytoskeletal rearrangement, while its overexpression promoted proliferation, enhanced F-actin filamentous structures, and strengthened cell–cell contacts. Collectively, our findings establish FAM65B as a novel positive regulator of SC function in chickens. These cellular findings, together with the association between FAM65B expression and semen quality traits, provide a foundation for further investigation of SC biology in avian male reproduction.

Cells doi: 10.3390/cells15211942

Authors: Polina S. Ermakova Anna A. Barinova Alexandra Y. Bogomolova Arseniy L. Potapov Polina M. Zharova Maxim A. Baten’kin Mariya Yu. Zakharina Alexey N. Konev Sergey A. Chesnokov Maria M. Karabut Vadim V. Elagin Alexandra N. Bogomazova Aleksandra V. Kashina Alexandra V. Panova Elena V. Zagaynova

Allogeneic cell replacement therapy for diabetes using induced pluripotent stem cell (iPSC)-derived pancreatic progenitors (PPs) is currently limited by immune rejection. Alginate encapsulation and the genetic ablation of HLA-I via β2-microglobulin (B2M) knockout represent distinct strategies to reduce immune recognition and to protect the transplanted cells. We generated encapsulated B2M-knockout pancreatic progenitors (IPS-∆B2M-PPs) and evaluated their immunogenicity in vitro using each of: peripheral blood mononuclear cell (PBMC) co-culture, natural killer cell (NK cell) degranulation, T-cell activation, and rat macrophage adhesion. In vivo studies were also performed to assess capsule biocompatibility and structural stability, following transplantation into rat omentum. While the B2M knockout did not compromise differentiation to pancreatic lineage, encapsulation largely prevented NK cell degranulation regardless of the encapsulated cell type, although T-cells did show residual activation after 7 days of co-culture with encapsulated progenitors, albeit significantly reduced in comparison with non-encapsulated controls. IPS-∆B2M-PP-containing capsules showed reduced macrophage adhesion in vitro compared to wild-type progenitors, while Interleukin-1β (IL-1β) secretion by the macrophages remained negligible. In vivo, all capsules formed a myofibroblastic zone without giant foreign-body cells; IPS-∆B2M-PP implants exhibited the thinnest fibrotic capsules and minimal lymphocytic infiltration. No capsule deformation was observed at 2 weeks or 1 month, indicating preserved structural stability. Although this characterizes both the local tissue response to the biomaterial and capsule stability, it does not establish immune evasion or reduced immunogenicity of the encapsulated cells in vivo. Overall, this study demonstrates the feasibility of combining B2M knockout with alginate encapsulation and suggests a potential additional protective role for B2M deletion under conditions of compromised capsule integrity that requires further investigation.

Cells doi: 10.3390/cells15211941

Authors: Mohammad Golam Sabbir Karin Hsiao

Mitochondrial dysfunction contributes to Alzheimer’s disease (AD), but the roles of individual respiratory-chain proteins remain incompletely understood. We examined mitochondrially encoded cytochrome c oxidase subunit II (MT-CO2) and nuclear encoded succinate dehydrogenase complex iron-sulfur subunit B (SDHB) in postmortem temporal cortex from cognitively normal (CN) individuals and patients with AD, Lewy body dementia/Parkinson’s disease (LBD/PD), or progressive supranuclear palsy (PSP). MT-CO2 abundance was lower in AD than in CN and LBD/PD, whereas SDHB was lower in AD than in CN, LBD/PD, and PSP. A significant protein × diagnosis interaction indicated distinct diagnostic-group patterns for MT-CO2 and SDHB. In BE(2)-C neuroblastoma cells, repeated 650 nm red-light and 808 nm near-infrared (NIR) photobiomodulation increased MT-CO2 abundance, with the largest responses following 10- and 15-min exposures. shRNA-mediated MT-CO2 reduction impaired Complex IV-dependent respiration and attenuated the bioenergetic response to 808 nm treatment. Photobiomodulation did not significantly alter mitochondrially encoded MT-CO1 or MT-CO3 abundance. These findings identify alterations in MT-CO2 and SDHB in AD temporal cortex and demonstrate that MT-CO2 contributes to Complex IV-dependent respiration and photobiomodulation-associated bioenergetic enhancement. Because mitochondrial content was not independently measured, the tissue findings cannot distinguish broader mitochondrial loss from protein-specific dysregulation.

Cells doi: 10.3390/cells15211940

Authors: Julia Niebora Julia Spaczyńska Aleksandra Partyńska Natalia Penciak Dominika Domagała Krzysztof Data Paul Mozdziak Artur Bryja Piotr P. Chmielewski Paweł Dąbrowski Teresa Wysocka Maciej Zabel Marzenna Podhorska-Okołów Piotr Dzięgiel Bartosz Kempisty Ievgeniia Kocherova

Ovarian cancer is one of the deadliest gynecological cancers worldwide. One promising therapeutic approach is the use of combined biomarkers. The Risk of Ovarian Malignancy Algorithm (ROMA) which combines CA125 with Human Epididymal Protein 4 (HE4) is an example of combining biomarkers to gain an optimal predictive outcome. Tumor Protein p53 (TP53) is a transcription factor with tumor suppressor properties. Mutations in TP53 constitute the most frequent genetic alterations observed in ovarian cancer. In contrast to conventional markers such as CA125, circulating tumor DNA (ctDNA) provides information about tumor mutations and insights into treatment response. Analysis of ctDNA enables the identification of tumor-associated mutations. Non-coding RNAs (ncRNAs) also influence cellular processes and act as effectors in oncogenic pathways by binding to RNA or proteins. One effector is Metastasis-Associated Lung Adenocarcinoma Transcript 1 (MALAT1), whose increased expression is associated with an aggressive tumor phenotype. MicroRNAs (miRNAs) are crucial in regulating the expression of genes associated with ovarian cancer development and progression. Exosomes carrying various molecules, including miRNAs and lncRNAs, contribute to ovarian cancer progression. Adipocytokines play an important role in the regulation of metabolic and inflammatory processes. Despite their diagnostic potential, the identified biomarkers require further validation.

Cells doi: 10.3390/cells15211939

Authors: Yu Ou Pucheng Wu Bingkun Xu Sheng Wang Shengxiao Cao Yuhan Sun Zixin Zhang Jinshuo Song Xue Sun Ning Qin Rifu Xu

Follicle-stimulating hormone (FSH) supports granulosa cell survival during follicular development, but the RNA networks involved remain incompletely understood. To investigate these networks, we integrated whole-transcriptome profiling and functional assays in cultured granulosa cells from chicken prehierarchical follicles. Transcriptomic comparisons included control, FSH and FSH plus leptomycin B (LMB), a nuclear export inhibitor. Integrated analysis of these profiles identified a candidate circARHGAP21/gga-miR-146a-5p/DEAF1 regulatory module. FSH increased circARHGAP21 and DEAF1 expression and decreased gga-miR-146a-5p abundance, whereas LMB opposed these changes in FSH-treated cells. Reporter assays showed that gga-miR-146a-5p suppressed reporter activity through the predicted sites in circARHGAP21 and the DEAF1 3′ untranslated region. Transfection with the oe-circARHGAP21 construct reduced apoptosis and increased follicle-stimulating hormone receptor (FSHR) expression. Construct transfection also increased proliferating cell nuclear antigen (PCNA) expression and decreased caspase-3 (CASP3) expression. Cotransfection with a gga-miR-146a-5p mimic attenuated these effects, whereas DEAF1 overexpression produced changes similar to those observed after oe-circARHGAP21 construct transfection. Compared with the oe-circARHGAP21 construct group, the corresponding LMB-treated group showed lower DEAF1, FSHR and PCNA expression and higher CASP3 expression and apoptosis. Together, these findings identify an FSH-responsive circARHGAP21/gga-miR-146a-5p/DEAF1 module associated with granulosa cell survival and follicle-selection-related gene expression.

Cells doi: 10.3390/cells15211938

Authors: Margarita Maiak Anna Efremova Tatiana Budagova Diana Mokrousova Dmitry Goldshtein

Adoptive T-cell therapy has demonstrated substantial clinical efficacy in several hematological malignancies, but autologous manufacturing remains limited by variability in starting cell material, manufacturing time, product heterogeneity, and T-cell dysfunction. Induced pluripotent stem cells (iPSCs) provide a potentially scalable and genetically engineerable source for the generation of standardized allogeneic T-cell products. This review summarizes current approaches to the differentiation of iPSCs into distinct T-cell phenotypes and evaluates their potential for cancer immunotherapy. We first discuss iPSC-derived conventional αβ T cells, including recent advances in feeder-free and scalable differentiation, maturation, and genetic engineering, as well as preclinical and early clinical evidence. We then examine γδ T cells, invariant natural killer T cells, and mucosal-associated invariant T cells, focusing on their distinctive antigen-recognition mechanisms, effector functions, tissue tropism, differentiation strategies, and therapeutic applications. Across these populations, iPSC technology may facilitate standardized and scalable production while enabling targeted engineering of T-cell specificity and functional properties. However, important challenges remain, including efficient lineage-specific differentiation, maturation, phenotypic consistency, long-term persistence, tumor infiltration, and control of alloreactivity and immunogenicity. Overall, iPSC-derived T cells represent a versatile platform for developing diverse cellular immunotherapies, although their clinical potential remains dependent on further technological and translational advances.

Cells doi: 10.3390/cells15211937

Authors: Gao-Xiang Ham George J. Augustine

Although the claustrum (CLA) has been increasingly implicated in salience and attentional processing, its relationship with the basolateral amygdala (BLA)—a key structure for emotional valence—remains poorly defined. Here, we used viral tracing, optogenetics, and brain slice electrophysiology to characterize the spatial organization and function of BLA–CLA circuitry. We found a clear anatomical compartmentalization of axonal projections between the BLA and CLA within the ring-like outer shell of the CLA, with little axonal labeling in the CLA core. Given this anatomical segregation, it was surprising that optogenetic photostimulation of BLA axons produced comparable excitatory responses in neurons in both the CLA core and shell. Pharmacological experiments indicated that BLA axons make monosynaptic contact with both shell and core neurons, with the latter likely doing so via their distal dendrites that extend into the shell. When CLA neurons were stratified according to their projection targets, BLA axons overlapped more with CLA neurons projecting to the motor cortex than those projecting to the retrosplenial cortex. However, synaptic responses to BLA input were similar in CLA neurons projecting to both cortical targets. Our results provide the first functional characterization of a BLA input pathway that potentially carries valence-related signals to the CLA. Because the excitatory synaptic responses produced in CLA neurons by BLA input were small and subthreshold for action potential generation, our results also argue that the BLA input contributes to a broader convergence of excitatory inputs instead of being a primary driver of CLA activity.

Cells doi: 10.3390/cells15211935

Authors: Baojun Liu Gang Liu Jianyu Bai Yujia Zhang Shu Wang Aixing Gu W. G. Dilantha Fernando

Cotton, as an economically important crop, suffers significant yield losses from infection by Fusarium oxysporum f. sp. vasinfectum race 7 (Fov7) infection. To elucidate the molecular mechanisms underlying host responses to Fov7 infection, we conducted a comprehensive transcriptional analysis of Gossypium barbadense using integrated bulk RNA sequencing (RNA-seq) and single-nucleus RNA sequencing (snRNA-seq) approaches. Our time-course study encompassed four critical infection stages (0, 2, 12, and 48 h post-inoculation) to characterize spatiotemporal transcriptomic dynamics. Bulk RNA-seq analysis revealed that the number of differentially expressed gene (DEG) numbers peaked at 2 h post-inoculation (hpi), identifying 9230 DEGs and demonstrating substantial transcriptional activation during early pathogen challenge. K-means clustering classified DEGs into six temporal expression patterns, with cluster C4 showing significant enrichment of defense-related pathways. Weighted gene co-expression network analysis (WGCNA) identified ten distinct co-expression modules, among which the turquoise module exhibited strong correlation with early infection responses. Integration of K-means clustering and WGCNA results pinpointed 1503 key defense-related genes. A single-nucleus atlas of cotton roots during Fov7 infection was constructed, comprising 32,773 nuclei, which revealed the cellular heterogeneity of cotton root tips. Functional enrichment analysis using these key genes emphasized the crucial roles of root cap and epidermal cells in pathogen perception and defense activation. Pseudotime trajectory mapping further delineated transcriptional dynamics in these cell populations during infection progression, demonstrating pathway activation in the root cap and epidermis at later infection stages. This multi-omics study establishes a valuable transcriptional resource for dissecting cotton’s resistance mechanisms against Fov7 and provides theoretical insights for developing disease-resistant cultivars through molecular breeding strategies.

Cells doi: 10.3390/cells15211934

Authors: Xiaofei Duan Yiqing Lu Hongyu Zhou Zi Zhang Zhanan Zhou Miao Wang Zhenna Chen Xiaoling Dun Yang Zhu Hanzhong Wang Lixi Jiang

Chemotherapy treatment of colorectal cancer (CRC) with cisplatin (CDDP) faces drug resistance and cytotoxicity issues, highlighting the urgent need for combination strategies. Selenium-enriched rapeseed extracts exhibit anticancer effects; however, their bioactive components and mechanisms remain unclear. Here, we applied different solvents to fractionate extracts of selenium-enriched rapeseed and found that the water extract (WE) fraction significantly enhanced the cytotoxic effect of CDDP on cancer cells without damaging normal cells, namely, HK-2 and NCM460. High-performance liquid chromatography–inductively coupled plasma–mass spectrometry (HPLC-ICP-MS) analysis revealed methylselenocysteine (MSC) and selenocysteine (SeCys2) as the main selenium species in the WE. Through cell biology and integrative multi-omics analysis, we found a synergistic anti-CRC cell effect upon combining CDDP with MSC, sulforaphane (SFN), celastrol (Cel), indole-3-carbinol (I3C), α-linolenic acid (ALA) or linoleic acid (LA). We propose that CDDP-WE (cisplatin and water extract) combination treatment holds promise for improving antitumor efficacy against chemo-refractory cells.

Cells doi: 10.3390/cells15211936

Authors: Shiqiao Ye Rabina Shrestha Yang Yu Matthew Alonzo Cankun Wang Qin Ma Ming-Tao Zhao

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) show decreasing cell-cycle activity during prolonged culture. However, it remains unclear whether changes in cell-cycle activity occur before, after, or concurrently with transcriptional changes in cell-cycle and maturation-associated programs. Here, we combined a TNNT2-FUCCI reporter, Ki67 immunostaining, and bulk RNA sequencing to define how cell-cycle activity changes during prolonged human iPSC-cardiomyocyte culture. FUCCI and Ki67 analyses showed similar levels of cell-cycle activity at days 20 and 30 of cardiomyocyte differentiation but declined more clearly by day 40. In contrast, transcriptomic profiling revealed that canonical cell-cycle and DNA-replication programs were already reduced by day 30. The subsequent day-30-to-day-40 interval was characterized by increased expression of sarcomeric, contractile, and calcium-handling gene programs rather than a further broad loss of mitotic pathways. These findings support a staged model of human iPSC-cardiomyocyte maturation-associated transcriptional remodeling in which a reduction in cell-cycle-associated transcription is detectable before the subsequent decline in cardiomyocyte cell-cycle activity, thereby identifying distinct temporal patterns of cell-cycle and maturation-associated changes during prolonged hiPSC-CM culture.

Cells doi: 10.3390/cells15211933

Authors: Sung Woo Cho Gil Won Lee Pil Soo Sung

Immunoglobulin A (IgA) connects mucosal defense with hepatic immune regulation, but its effects vary by anatomical compartment, antibody properties, and responding cell type. This narrative review synthesizes experimental and clinical evidence on the gut–liver IgA axis in alcohol-associated liver disease, metabolic dysfunction-associated steatotic liver disease and steatohepatitis, and hepatocellular carcinoma. We distinguish IgA-producing cells from secreted antibodies and IgA-responsive cells and examine antibody production, cellular trafficking, hepatic clearance, and biliary export. Experimental studies demonstrate the protective effects of mucosal IgA delivery and the context-dependent contributions of IgA-associated responses to inflammation, fibrosis, and impaired antitumor immunity. Human studies identify tissue and prognostic associations but do not establish a common intestinal source of pathogenic hepatic IgA. Collectively, these findings provide a rationale for investigating disease-specific, compartment-directed approaches targeting antibody production, cellular trafficking, protective export, or local effector responses. Clinical translation requires validation of causal mechanisms and intervention-specific biomarkers, as well as evidence of improved liver outcomes while preserving mucosal protection.

Cells doi: 10.3390/cells15211932

Authors: Eugenia Rubino Abraham Tettey-Matey Alice Giusto Francesca Sbrana Margherita Festa Maria Antonietta Coppola Cristiana Picco Paola Gavazzo Elena Angeli Michael Pusch Raffaella Barbieri

Endosomal ClC-3 and ClC-4 Cl−/H+ antiporters are essential regulators of vesicular ion homeostasis and neuronal function, yet the mechanisms controlling their activity remain incompletely understood. TMEM9A has recently emerged as an accessory β-subunit of endosomal CLC transporters. Here, we define the functional and molecular specificity of TMEM9A across the CLC family. Electrophysiological recordings in Xenopus laevis oocytes and HEK293 cells demonstrated that TMEM9A strongly suppresses ClC-3- and ClC-4-mediated transport, while producing no detectable inhibition of ClC-2 or ClC-7 and only a modest effect on ClC-1. Consistent with these functional effects, colocalization microscopy and FLIM-FRET revealed close molecular association of TMEM9A with ClC-3 and ClC-4, with FRET efficiencies of 0.19 ± 0.02 and 0.15 ± 0.02, respectively, whereas substantially weaker interactions were detected with ClC-1 and ClC-7. In the mouse brain, Tmem9a, Tmem9b, Clcn3, and Clcn4 displayed broad and overlapping expression across cortex, hippocampus, and cerebellum. Moreover, immunofluorescence analysis showed moderate colocalization of TMEM9A with ClC-3 in primary neurons and substantial colocalization with ClC-3 and ClC-4 in heterologous cells. Together, these findings establish a preferential functional and molecular association of TMEM9A with ClC-3 and ClC-4, closely mirroring the selectivity of TMEM9B, and support the TMEM9 proteins as a regulatory β-subunit system for the neuronal endosomal CLC transporters.

Cells doi: 10.3390/cells15211931

Authors: Marianna Weener Konstantin Kotliar

Vertebrate rod and cone photoreceptors are among the most morphologically extreme cells in human biology, resembling very thin sticks with elongated outer segments connected to the inner segments by a narrow connecting cilium (CC) of approximately 200–300 nm in diameter. This specialized organelle serves as the sole conduit for the massive daily protein and membrane trafficking required for outer segment renewal, while simultaneously providing structural anchorage. Dysfunction of the CC and its associated molecular machinery, most directly the failure of protein and lipid trafficking across the cilium, underlies numerous retinal ciliopathies, including retinitis pigmentosa (RP) and Leber congenital amaurosis (LCA). Recent ultrastructural studies have revealed an inner protein scaffold within the CC comprising FAM161A, POC5, and centrin that acts as a “structural zipper” maintaining microtubule cohesion. Despite these advances, the mechanical properties of the CC remain uncharacterized, and whether biomechanical compromise of the CC contributes to ciliopathy progression is unknown. This review critically synthesizes current knowledge of photoreceptor CC structure, biomechanics, and dysfunction; evaluates existing and repurposed pharmacological agents that may protect photoreceptors through ciliary or non-ciliary mechanisms; and assesses the speculative but intriguing concept of exogenous organelle-scale mechanical stabilization. We identify critical gaps at the intersection of mechanobiology, nanobiomaterials, and retinal therapeutics, proposing a research agenda for testing whether biomechanical insights can be translated into clinically actionable strategies.