International Journal of Molecular Sciences

13 pages, 3306 KB

Open AccessCase Report

Intrafamilial Variability in NRXN1-Associated Neurodevelopmental Disorders: Clinical and Genetic Insights from a Family Case Study with Literature Review

by Nikolina Kastratovic, Marina Gazdic Jankovic, Marina Miletic Kovacevic, Sandra Nikolic, Dragica Pavlovic, Dijana Perovic, Vladimir Janjic and Biljana Ljujic

Int. J. Mol. Sci. 2026, 27(14), 6241; https://doi.org/10.3390/ijms27146241 (registering DOI) - 13 Jul 2026

Abstract

The neurexin1 gene (NRXN1) encodes a presynaptic adhesion molecule that plays a critical role in synapse formation, maintenance, and function. Copy-number variants (CNVs) affecting the NRXN1 locus, including submicroscopic deletions, represent rare variant acting as a predisposition for neurodevelopmental disorders, such [...] Read more.

The neurexin1 gene (NRXN1) encodes a presynaptic adhesion molecule that plays a critical role in synapse formation, maintenance, and function. Copy-number variants (CNVs) affecting the NRXN1 locus, including submicroscopic deletions, represent rare variant acting as a predisposition for neurodevelopmental disorders, such as Pitt–Hopkins-like syndrome type 2 (MIM #614325) and susceptibility to schizophrenia (MIM #621407). Variations in NRXN1 gene are associated with marked clinical heterogeneity. We present a familial case involving two male siblings (aged 6 and 5 years) and their 28-year-old mother, all exhibiting variable neurodevelopmental phenotypes. Both children demonstrated disharmonic developmental profiles characterized by impaired communication, speech largely intelligible only to their parents, and behaviors consistent with autism spectrum disorder, including reduced eye contact. Mother represents a carrier with only subtle, nonspecific behavioral traits, further supporting the concept of incomplete penetrance and variable expressivity associated with this genetic alteration. Genetic analysis identified a 317 kb NRXN1 deletion shared by all affected family members, accompanied by significant intrafamilial phenotypic variability, suggesting the contribution of additional genetic and/or modifying factors. These findings support the concept that NRXN1 deletions alone do not determine clinical outcome but rather act within a broader genetic and biological context. The marked intrafamilial phenotypic variability and incomplete penetrance observed in this family is compatible with a multiple-hit model, whereby NRXN1 deletions act as susceptibility factors whose phenotypic consequences are shaped by additional genetic and modifying influences. However, the genetic mechanisms underlying the observed phenotypic variability warrant further investigation. Full article

(This article belongs to the Special Issue Molecular Investigations in Neurodevelopmental Disorders: 2nd Edition)

22 pages, 922 KB

Open AccessArticle

Volatilomic Signatures of Parental and Oxaliplatin-Resistant HCT116 Colon Cancer Cell Lines

by Christine Heinzle, Andreas Leiherer, Axel Muendlein, Clemens Ager, Agnieszka Królicka, Chris A. Mayhew and Pawel Mochalski

Int. J. Mol. Sci. 2026, 27(14), 6240; https://doi.org/10.3390/ijms27146240 (registering DOI) - 13 Jul 2026

Abstract

Volatile organic compounds (VOCs) reflect cellular metabolic activities and may serve as non-invasive biomarkers in oncology. This study investigated whether acquired oxaliplatin resistance in colorectal cancer is associated with distinct volatilomic alterations. The human colorectal cancer cell line HCT116 and its oxaliplatin-resistant derivative [...] Read more.

Volatile organic compounds (VOCs) reflect cellular metabolic activities and may serve as non-invasive biomarkers in oncology. This study investigated whether acquired oxaliplatin resistance in colorectal cancer is associated with distinct volatilomic alterations. The human colorectal cancer cell line HCT116 and its oxaliplatin-resistant derivative (OXrHCT116) were analyzed under basal conditions and following oxaliplatin exposure. Chemoresistance was confirmed using dose–response, cell viability, and colony formation assays. VOCs were analyzed by headspace needle trap extraction coupled with gas chromatography–mass spectrometry (HS-NTE-GC-MS). OXrHCT116 cells exhibited markedly reduced oxaliplatin sensitivity, increased IC₅₀ values, and reduced proliferative and clonogenic capacity. Volatilomic profiling identified 55 significantly altered VOCs. Parental HCT116 cells displayed broader VOC diversity and higher turnover than OXrHCT116 cells. Hydrocarbons associated with lipid peroxidation and oxidative stress were more abundant in HCT116 cells, whereas resistant cells showed markedly reduced emission of these compounds. Additional alterations in aldehydes, alcohols, and aromatic compounds suggested reduced metabolic flux in resistant cells. Oxaliplatin exposure induced pronounced volatilomic changes in HCT116 cells but only minimal modulation in OXrHCT116 cells. These findings suggest that oxaliplatin resistance may be associated with distinct metabolic reprogramming and support VOC profiling as a promising approach for monitoring chemoresistance. Full article

(This article belongs to the Special Issue Machine Learning and Bioinformatics in Human Health and Disease: 2nd Edition)

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15 pages, 1226 KB

Open AccessArticle

Identification of RhoGAP Gene Family in Soybean (Glycine max L.) and its Role in the Response to Rhizobium Infection

by Chengcheng Qin, Han Huang, Yanbo Sun, Ruixue Luo, Xin Zhang, Bohong Su and Jian Song

Int. J. Mol. Sci. 2026, 27(14), 6239; https://doi.org/10.3390/ijms27146239 (registering DOI) - 13 Jul 2026

Abstract

Rho GTPase-activating proteins (RhoGAPs) are characterized by a conserved RhoGAP domain and function as negative regulators of Rho GTPases, playing important roles in plant growth, development, and responses to environmental stimuli. In this study, 19 GmRhoGAP genes were identified in the soybean genome [...] Read more.

Rho GTPase-activating proteins (RhoGAPs) are characterized by a conserved RhoGAP domain and function as negative regulators of Rho GTPases, playing important roles in plant growth, development, and responses to environmental stimuli. In this study, 19 GmRhoGAP genes were identified in the soybean genome and found to be unevenly distributed across 11 chromosomes. Comprehensive analyses were performed, including gene structure, conserved motifs, protein domains, gene duplication, synteny, cis-acting elements, tissue-specific expression, and quantitative real-time PCR under rhizobial infection. Structural analysis revealed substantial diversity in intron-exon organization but high conservation of motif composition, with all members containing the conserved RhoGAP domain. A total of 20 segmentally duplicated gene pairs were identified, indicating expansion of the GmRhoGAP family in soybean. Inter-species synteny analysis showed closer evolutionary relationships with Arabidopsis than with rice, and Ka/Ks analysis suggested strong purifying selection during evolution. Promoter analysis indicated potential involvement in development, phytohormone signaling, stress responses, and light responsiveness. Expression profiling demonstrated distinct tissue-specific patterns. qRT-PCR further showed that GmRhoGAP genes respond differentially to rhizobial infection, with 10 generally upregulated genes and 4 downregulated genes. These analyses provide a framework for understanding this gene family and identify candidate genes for future research. Full article

(This article belongs to the Special Issue Latest Research on Plant Genomics and Genome Editing, 2nd Edition)

21 pages, 4304 KB

Open AccessArticle

Evaluating KRAS-Associated Responses to Sulfasalazine and 5-Fluorouracil in Colorectal Cancer Using Integrated 2D and PEGDA Microwell-Based 3D Tumor Models

by Mehrdad Bandegi, Ezgi Biltekin, Yasemin M. Akay and Metin Akay

Int. J. Mol. Sci. 2026, 27(14), 6238; https://doi.org/10.3390/ijms27146238 (registering DOI) - 13 Jul 2026

Abstract

Colorectal cancer (CRC) is a major cause of cancer-related mortality among adults younger than 50 years of age, and many tumors show incomplete response or develop resistance to 5-fluorouracil (5-FU)-based chemotherapy. Therefore, new therapeutic approaches that improve CRC sensitivity to existing chemotherapeutic agents [...] Read more.

Colorectal cancer (CRC) is a major cause of cancer-related mortality among adults younger than 50 years of age, and many tumors show incomplete response or develop resistance to 5-fluorouracil (5-FU)-based chemotherapy. Therefore, new therapeutic approaches that improve CRC sensitivity to existing chemotherapeutic agents are needed. KRAS-associated signaling contributes to CRC growth, metabolic adaptation and treatment resistance. In this study, we investigated whether sulfasalazine (SSZ), a U.S. Food and Drug Administration (FDA)-approved anti-inflammatory drug, could enhance the response of CRC cells to 5-FU and modulate KRAS/mitogen-activated protein kinase (MAPK)-associated signaling. Public dataset analysis using cBioPortal, Kaplan–Meier Plotter and DepMap showed that KRAS is commonly altered in CRC. The analysis also showed that higher KRAS expression was associated with shorter overall survival in 1061 CRC patients, while CRC cell-line models demonstrated KRAS dependency. In 2D cultures, both KRAS-mutant HCT116 and KRAS-wild-type RKO cells showed lower cell viability, reduced colony formation and decreased KRAS expression after SSZ treatment. In 3D cultures, exposure to SSZ reduced early spheroid formation, both as a single treatment and when combined with 5-FU. In established spheroids, SSZ-containing treatments affected cell viability, spheroid growth and morphology, with the most noticeable suppressive effect observed in RKO aggregates. SynergyFinder+ dose-matrix analysis identified dose ranges where SSZ and 5-FU showed additive-to-synergistic effects, leading us to select 600 μM SSZ with 25 μM 5-FU for further validation. Western blot results from PEGDA microwell-derived 3D spheroids showed that SSZ + 5-FU treatment reduced KRAS expression and affected KRAS/MAPK-related signaling. This effect was more pronounced in RKO cells, where downstream pathway suppression was stronger. The combination treatment also increased apoptosis-associated PARP cleavage. At the same time, it reduced Cyclin D1 and GPX4 protein levels and changed the expression of stemness-related markers, including ALDH1A3, CD44, and CD133. Together, these results support SSZ as a candidate repurposed adjuvant that may improve the response to 5-FU in CRC spheroid models and support the use of PEGDA microwell-based 3D platforms for testing combination therapy approaches. Full article

(This article belongs to the Special Issue Novel Therapeutic Targets in Cancers: 5th Edition)

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19 pages, 1354 KB

Open AccessArticle

Juglone/MWCNT-Modified Electrode for High-Performance Melatonin Detection

by Joanna Smajdor-Baran

Int. J. Mol. Sci. 2026, 27(14), 6237; https://doi.org/10.3390/ijms27146237 (registering DOI) - 13 Jul 2026

Abstract

The integration of carbon nanomaterials with organic compounds offers a promising strategy for developing next-generation electrode materials with superior properties. A novel type of carbon paste electrode was fabricated by modifying a graphite matrix with functionalized multiwalled carbon nanotubes and juglone as a [...] Read more.

The integration of carbon nanomaterials with organic compounds offers a promising strategy for developing next-generation electrode materials with superior properties. A novel type of carbon paste electrode was fabricated by modifying a graphite matrix with functionalized multiwalled carbon nanotubes and juglone as a redox mediator, and then it was deposited by manual packing into a PEEK body (JUG-MWCNT/CPE). The surface morphology and structural parameters of the composite materials were meticulously characterized using scanning electron microscopy (SEM), nitrogen adsorption–desorption isotherms, and spectroscopic techniques, while their electrochemical properties were rigorously evaluated using cyclic voltammetry (CV) and differential pulse voltammetry (DPV). This study demonstrates that the synergistic interaction between the conductive nanotube network and the electroactive juglone significantly reduces the oxidation overpotential and enhances the peak current response of melatonin. Under optimized DPV parameters, the developed sensor presents outstanding analytical performance, featuring a wide linear response range from 0.002 to 0.16 mg L⁻¹, a low detection limit of 0.49 µg L⁻¹, excellent long-term signal stability for up to 30 days, and valid applicability for real-sample monitoring in commercial tablets and dietary supplements. Full article

(This article belongs to the Special Issue Electrochemical Detection: A Molecular-Level Perspective)

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22 pages, 1879 KB

Open AccessArticle

Multi-Omics and Experimental Validation Reveal the Protective Effect of Paeoniflorin Against Coronary Heart Disease in Mice via Inhibiting the C3-Cfd-C3aR Pathway

by Ying Yang, Xiang Li, Wenjing Zong, Sijia Wu, Yingying Li, Danli Tang and Huamin Zhang

Int. J. Mol. Sci. 2026, 27(14), 6236; https://doi.org/10.3390/ijms27146236 (registering DOI) - 13 Jul 2026

Abstract

Coronary heart disease (CHD) is a global cardiovascular disease with high morbidity and mortality, and its complex pathological mechanism poses great challenges to clinical prevention and treatment. Paeoniflorin (PA), a monoterpene glycoside active ingredient from Ranunculaceae plants, has shown potential in cardiovascular protection, [...] Read more.

Coronary heart disease (CHD) is a global cardiovascular disease with high morbidity and mortality, and its complex pathological mechanism poses great challenges to clinical prevention and treatment. Paeoniflorin (PA), a monoterpene glycoside active ingredient from Ranunculaceae plants, has shown potential in cardiovascular protection, but its specific anti-CHD molecular targets and systematic regulatory networks remain unclear. In this study, a mouse model of CHD was established, and a multi-omics strategy combining label-free quantitative proteomics and metabolomics was adopted to explore the mechanism of PA in treating CHD. The results showed that PA significantly improved cardiac function, alleviated myocardial pathological injury and fibrosis, and regulated lipid metabolism in CHD model mice, with the high-dose group showing the optimal effect. Proteomic analysis identified 51 key differentially expressed proteins (DEPs) reversed by PA, which were mainly enriched in complement and coagulation cascades, and neutrophil extracellular trap formation pathways, with the C3-Cfd-C3aR signaling axis as the core hub. Further verification confirmed that PA could downregulate the expression of C3, Cfd, C3aR, and their downstream molecule BTK, thereby inhibiting myocardial inflammatory response and cardiomyocyte apoptosis. In addition, PA downregulated the expression of platelet activation markers ITGA2B/ITGB3. Metabolomic analysis revealed that PA reversed 57 abnormal metabolites in CHD mice, which were enriched in GABAergic synapse, retrograde endocannabinoid signaling and other pathways. Molecular docking confirmed that PA could stably bind to C3, Cfd, C3aR, BTK, and ITGA2B/ITGB3 with strong binding activity. In conclusion, PA exerts anti-CHD effects through a multi-target and multi-pathway synergism, mainly by targeting the C3-Cfd-C3aR axis to inhibit inflammation, apoptosis and platelet activation, and regulating metabolic disorders. This study provides experimental evidence and theoretical support for the clinical application of PA as a multi-target therapeutic drug for CHD. Full article

(This article belongs to the Special Issue Bioactive Natural Products: From Molecular Mechanisms to Biological Function)

13 pages, 1165 KB

Open AccessArticle

Synthesis of Silver Phosphate (Ag₃PO₄) and Its Supplementation to Enhance Phytochemical Accumulation and Antioxidant Activity of Cryptocoryne albida Under Hydroponic Cultivation

by Somkiat Seesanong, Banjong Boonchom, Chaowared Seangarun, Nongnuch Laohavisuti, Benyakorn Labantao, Pesak Rungrojchaipon, Sirichet Punthipayanon and Wimonmat Boonmee

Int. J. Mol. Sci. 2026, 27(14), 6235; https://doi.org/10.3390/ijms27146235 (registering DOI) - 13 Jul 2026

Abstract

This study investigated the supplementation of silver phosphate (Ag₃PO₄) to enhance the growth and antioxidant activity of Cryptocoryne albida in a hydroponic system. Ag₃PO₄ was synthesized via a simple precipitation method using AgNO₃ and (NH [...] Read more.

This study investigated the supplementation of silver phosphate (Ag₃PO₄) to enhance the growth and antioxidant activity of Cryptocoryne albida in a hydroponic system. Ag₃PO₄ was synthesized via a simple precipitation method using AgNO₃ and (NH₄)₂HPO₄ as precursors, with alcohol employed as the reaction medium to control particle growth and agglomeration. FTIR analysis confirmed the presence of characteristic phosphate (PO₄³⁻) functional groups, while XRD revealed diffraction peaks consistent with Ag₃PO₄ as the dominant crystalline phase. The SEM image exhibits relatively uniform spherical particles of 300–500 nm. Moreover, the XRF results for the synthesized Ag₃PO₄ indicated 86.80% of Ag₂O and 12.60% of P₂O₅ as the major components. The application experiment was conducted in a DFT hydroponic system containing different concentrations of Ag₃PO₄-derived supplementation (0.00, 0.01, 0.10, and 1.00 ppm). The highest concentration (1.00 ppm) of Ag₃PO₄ in the KMITL2 nutrient solution significantly reduced plant growth (p < 0.05). In contrast, Ag₃PO₄-derived supplementation with 0.01 ppm resulted in significantly higher phytochemical accumulation, including total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activities evaluated by DPPH and ABTS assays (p < 0.05). These results suggest that low-concentration Ag₃PO₄-derived supplementation may promote phytochemical accumulation and antioxidant activity in C. albida. Overall, this study highlights the potential of Ag₃PO₄-derived supplementation for modulating phytochemical accumulation and antioxidant activity in hydroponically cultivated C. albida. Full article

(This article belongs to the Special Issue Nanostructured Strategies for Bioactive Compounds)

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37 pages, 1266 KB

Open AccessReview

Membrane-Targeted Consequences of Acetaminophen Toxicity and Off-Target Effects of Antimicrobial Peptides on Host Cell Membranes

by Oksana M. Voloshchuk, Volodymyr Berest and Oleksii Skorokhod

Int. J. Mol. Sci. 2026, 27(14), 6234; https://doi.org/10.3390/ijms27146234 (registering DOI) - 13 Jul 2026

Abstract

Acetaminophen (paracetamol, APAP) is a widely used analgesic and antipyretic drug. Under normal physiological conditions, it does not directly interact with or disrupt cellular membranes. However, in cases of acetaminophen overdose or toxicity, severe cellular damage has been described, involving a broad spectrum [...] Read more.

Acetaminophen (paracetamol, APAP) is a widely used analgesic and antipyretic drug. Under normal physiological conditions, it does not directly interact with or disrupt cellular membranes. However, in cases of acetaminophen overdose or toxicity, severe cellular damage has been described, involving a broad spectrum of effects at different cellular levels. These toxic effects may involve membrane structures, including mitochondrial, plasma, and other intracellular membranes. Antimicrobial peptides (AMPs) are short, usually cationic and amphipathic peptides produced by both microorganisms and multicellular organisms, serving diverse defensive and competitive functions. In many cases, they exert their antimicrobial activity by direct interaction with bacterial or fungal membranes, leading to membrane destabilization and cell death. Owing to this membrane-targeting mechanism, AMPs may also interact with eukaryotic cell membranes, thereby exerting toxic or off-target effects under certain conditions. Here, we review the current knowledge on the membrane-related effects of acetaminophen toxicity and the mechanisms by which AMPs interact with biological membranes. In the event of combined exposure to acetaminophen and AMPs in therapeutic or experimental settings, the biological consequences remain unexplored. Such combined exposure may give rise to toxic effects and membrane-associated alterations. We further discuss potential mechanisms of interference, additive toxicity, and synergistic interactions between acetaminophen and AMPs, highlighting critical knowledge gaps and directions for future research. Full article

(This article belongs to the Special Issue Bioactive Molecules in Infection and Toxicity: Antimicrobial Peptides and Drug-Induced Damage in Disease Models)

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22 pages, 1964 KB

Open AccessArticle

Comparative Analysis of Endothelial Cell Culture Models Under Altered Mechanical Conditions and Glucose Variations

by Augusta Volkevičiūtė, Jayashree Sahana, Estéfano Pinilla, Daniela Melnik, Luis Fernando González-Torres, Markus Wehland, Edgaras Stankevicius, Daniela Grimm and Ulf Simonsen

Int. J. Mol. Sci. 2026, 27(14), 6233; https://doi.org/10.3390/ijms27146233 (registering DOI) - 13 Jul 2026

Abstract

Endothelial dysfunction is a defining feature of diabetic vascular disease and is characterized by impaired nitric oxide signaling, inflammatory activation, altered mechanotransduction, and disturbed angiogenic responses. The present study investigated whether hyperglycemia modulates endothelial phenotype in a model-dependent manner under distinct structural and [...] Read more.

Endothelial dysfunction is a defining feature of diabetic vascular disease and is characterized by impaired nitric oxide signaling, inflammatory activation, altered mechanotransduction, and disturbed angiogenic responses. The present study investigated whether hyperglycemia modulates endothelial phenotype in a model-dependent manner under distinct structural and mechanical culture conditions. EA.hy926 endothelial cells were cultured for 7 days under normoglycemic (5.5 mM) or high-glucose (25 mM) conditions as static monolayers (G-force = 1 g), adherent clinorotated cells, multicellular spheroids (MCSs) generated during clinorotation, and Matrigel-derived endothelial structures. Gene expression was assessed by qPCR using marker panels related to nitric oxide signaling, PI3K CA-AKT-mTOR signaling, inflammatory adhesion, angiogenesis, and structural adhesion, whereas protein abundance and spatial distribution of eNOS, AKT1, VCAM1, vinculin, and VEGFA together with CDH5/VE-cadherin were analyzed by Western blotting and confocal microscopy. Clinorotation generated both adherent endothelial cells and MCSs. High glucose reduced eNOS-related expression in most models, with the strongest decreases in adherent clinorotated cells and MCSs, whereas Matrigel cultures showed a divergent transcriptional response. PI3K CA expression was markedly suppressed by high glucose in clinostat-derived populations, while mTOR was differentially regulated in spheroids and Matrigel cultures. VCAM1 expression was most prominent in MCSs, whereas ICAM1 was highest in Matrigel cultures. VEGF-related signaling differed substantially among models, and Matrigel cultures showed the strongest VEGFA-associated protein signal and the clearest angiogenic organization under normoglycemic conditions, which became less distinct under high glucose. Vinculin protein abundance was highest in MCSs and Matrigel cultures, reflecting pronounced differences in structural organization. Overall, these findings show that endothelial responses to hyperglycemia are strongly shaped by mechanical and structural context and support the use of complementary in vitro models for studying diabetic endothelial dysfunction. Full article

(This article belongs to the Special Issue Microgravity, Cell Shape and Gene Expression: State of the Art, Challenges and Prospects)

17 pages, 7809 KB

Open AccessArticle

Toll-like Receptor 8 (TLR8) Is Expressed in Mouse Hippocampal Neurons and Modulates Neuronal Excitability

by Julian Fenkart, Alice Santagostino, Stephanie Seidlberger, Armin Schmuck, Katharina Donat, Meinrad Drexel and Sandra Santos-Sierra

Int. J. Mol. Sci. 2026, 27(14), 6232; https://doi.org/10.3390/ijms27146232 (registering DOI) - 13 Jul 2026

Abstract

Toll-like receptors (TLRs) are essential in the innate immune response. Furthermore, neuronal TLRs have been involved in regulating neuronal dendritic outgrowth and excitability. However, a deeper understanding of neuronal-TLR function is essential. We investigated the expression and role of TLR8 using mouse postnatal [...] Read more.

Toll-like receptors (TLRs) are essential in the innate immune response. Furthermore, neuronal TLRs have been involved in regulating neuronal dendritic outgrowth and excitability. However, a deeper understanding of neuronal-TLR function is essential. We investigated the expression and role of TLR8 using mouse postnatal hippocampal neuronal cultures. We assessed TLR8 (and TLR7) expression by immunocytochemistry and TLR8 functionality via electrophysiological recordings in a high-density multi-electrode array system and calcium flux imaging. Pathway activation following fast TLR8 stimulation was performed via Western blot. TLR8 and TLR7 are expressed in postnatal hippocampal neurons at DIV14 and in neurosphere-derived neurons. TLR8 expression was detected in the cell body and dendrites of GABAergic, parvalbumin-expressing interneurons. Stimulation with the TLR8 agonist TL8-506 led to increased neuronal activity in the short term and enhanced neuronal synchronization in short- and long-term recordings. The calcium flux induced by TLR8 was reduced by blocking NMDA receptors with D-AP5. TLR8 stimulation did not lead to IkB-α degradation. Our findings demonstrate that TLR8 and TLR7 are expressed in mouse postnatal hippocampal neurons. TLR8 activation leads to increased neuronal excitability that was decreased by NMDA antagonism. However, acute TLR8 stimulation failed to robustly activate the canonical TLR signaling pathway. We propose that TLR8 activation might lead to the enhancement of excitatory NMDA receptors. Full article

(This article belongs to the Special Issue The Role of Toll-Like Receptors (TLRs) in Infection and Inflammation: 4th Edition)

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18 pages, 15530 KB

Open AccessArticle

Study of Cytotoxicity of Pyrrolo[3,4-d]isoxazoline and Pyrrolo[2,1-a]isoquinoline Derivatives Against Tumor Cell Lines

by Andrew S. Drachuk, Sergey S. Mkrtchan, Stanislav V. Shmakov, Sergey Yu. Vyazmin, Kristina A. Kim, Alexander V. Stepakov and Vitali M. Boitsov

Int. J. Mol. Sci. 2026, 27(14), 6231; https://doi.org/10.3390/ijms27146231 - 13 Jul 2026

Abstract

Antiproliferative activity of pyrrolo[3,4-d]isoxazolines and pyrrolo[2,1-a]isoquinolines derived from them was studied against human erythroleukemia (K562), cervical carcinoma (HeLa), and melanoma (Sk-mel-2) cell lines in vitro by MTS assays followed by study of their effect on actin cytoskeleton and cell [...] Read more.

Antiproliferative activity of pyrrolo[3,4-d]isoxazolines and pyrrolo[2,1-a]isoquinolines derived from them was studied against human erythroleukemia (K562), cervical carcinoma (HeLa), and melanoma (Sk-mel-2) cell lines in vitro by MTS assays followed by study of their effect on actin cytoskeleton and cell motility by confocal microscopy, and apoptotic activity by flow cytometry. Most effective among the screened compounds were bicyclic hydroxylactams 6–9 with a pyrrolo[3,4-d]isoxazoline structure; they showed IC₅₀ values ranging from 12 to 36 μg/mL for all tested cancer cell lines with selectivity indexes up to 12 (as compared to the embryonic kidney HEK293T cell line). Loss of stress fibers with diffuse redistribution of granular actin throughout the cytoplasm in up to 25% of treated cells and a decrease in filopodia-like protrusions up to 69% were observed by confocal microscopy during an actin cytoskeleton study. Such cytoskeletal changes and the proposed altered cell motility were confirmed by scratch-test (revealed a three-fold decrease in cell motility). Full article

(This article belongs to the Special Issue Bioactive Compounds in Cancers: Second Edition)

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1 pages, 147 KB

Open AccessRetraction

RETRACTED: Ahmad et al. Deciphering the Potential Neuroprotective Effects of Luteolin Against Aβ₁–₄₂-Induced Alzheimer’s Disease. Int. J. Mol. Sci. 2021, 22, 9583

by Sareer Ahmad, Myeung Hoon Jo, Muhammad Ikram, Amjad Khan and Myeong Ok Kim

Int. J. Mol. Sci. 2026, 27(14), 6230; https://doi.org/10.3390/ijms27146230 - 13 Jul 2026

Abstract

The journal retracts the article titled “Deciphering the Potential Neuroprotective Effects of Luteolin against Aβ₁–₄₂-Induced Alzheimer’s Disease” [...] Full article

(This article belongs to the Special Issue Natural Products and Neuroprotection 3.0)

13 pages, 4526 KB

Open AccessCase Report

Evolution of Recurrent Myxofibrosarcoma of the Thoracic Wall at Single-Cell Resolution: A Case Report

by Elena E. Kopantseva, Artem L. Toropov, Timur I. Fetisov, Alexander V. Ikonnikov, Alexandra V. Sentyabreva, Maxim E. Menyailo, Anastasia A. Tararykova, Polina A. Shtompel, Victoria Y. Zinovieva, Kirill I. Kirsanov, Evgeny V. Denisov and Marianna G. Yakubovskaya

Int. J. Mol. Sci. 2026, 27(14), 6229; https://doi.org/10.3390/ijms27146229 - 13 Jul 2026

Abstract

Myxofibrosarcoma (MFS) is a common yet understudied type of soft tissue sarcoma. It is characterized by diverse cellular morphology, unusual growth patterns, and a propensity for local tumor recurrences. A 76-year-old patient underwent multiple surgical removals of MFS recurrences of the thoracic cavity [...] Read more.

Myxofibrosarcoma (MFS) is a common yet understudied type of soft tissue sarcoma. It is characterized by diverse cellular morphology, unusual growth patterns, and a propensity for local tumor recurrences. A 76-year-old patient underwent multiple surgical removals of MFS recurrences of the thoracic cavity and surrounding tissues with the subsequent development of metastases. Single-cell RNA sequencing was used to analyze the earlier (R7) and later (R8) MFS recurrences, with R8 removed less than a year before the detection of metastases in the lungs. The earlier MFS recurrence displays tumor clusters with multiple immunomodulatory markers (DKK1, APP, CD24, GRN) and genes involved in lipid metabolism and cell stress (DDIT3, ABCA1, ABCA10, ATF4, NEU1), combined with an anti-inflammatory TME. The later MFS recurrence shifts to a functionally less diverse phenotype, enriched in fibroblast-typical markers (POSTN, NES, COL1A1/A2), and a pro-inflammatory TME. The proliferative (MKI67, TOP2A, BUB1B) and mRNA splicing and processing (SNRNP70, SRSF2/5/11, RSRP1, LUC7L/7L3, SRRM1/2) tumor clusters are observed in both MFS recurrences, and their signatures match the previous data on primary MFS tumor populations. ScRNA-seq analysis of two subsequent MFS recurrences from one patient show a change from functionally diverse to more uniform ECM remodeling enriched tumor populations, an accompanying shift to the pro-inflammatory TME, and the presence of two common tumor clusters enriched in proliferative and mRNA-processing gene signatures. Full article

(This article belongs to the Special Issue Novel Therapeutic Targets in Cancers: 5th Edition)

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29 pages, 1730 KB

Open AccessReview

Advances in Immunotherapy for Intrahepatic Cholangiocarcinoma

by Huimin Qi, Jialin Pan and Hailong Wu

Int. J. Mol. Sci. 2026, 27(14), 6228; https://doi.org/10.3390/ijms27146228 - 13 Jul 2026

Abstract

Intrahepatic cholangiocarcinoma (iCCA) is a highly lethal and heterogeneous primary liver malignancy with a dismal prognosis. Approximately 70% of patients are diagnosed at locally advanced or metastatic stages, therefore missing the opportunity for curative surgery, and conventional chemotherapy offers limited survival benefits. Immunotherapy, [...] Read more.

Intrahepatic cholangiocarcinoma (iCCA) is a highly lethal and heterogeneous primary liver malignancy with a dismal prognosis. Approximately 70% of patients are diagnosed at locally advanced or metastatic stages, therefore missing the opportunity for curative surgery, and conventional chemotherapy offers limited survival benefits. Immunotherapy, especially immune checkpoint blockade, represents a promising strategy, yet its efficacy as monotherapy in iCCA remains modest primarily due to the profoundly immunosuppressive and desmoplastic tumor microenvironment. This review examines the immune cell infiltration landscape of iCCA, focusing on the distinct roles of lymphoid cells and myeloid cells in shaping immune evasion. We then analyze key factors affecting immune responses, such as tumor-intrinsic driver mutations, immune regulatory mechanisms, and acquired resistance. Furthermore, we summarize current clinical advances in iCCA immunotherapy, including immune checkpoint inhibitor monotherapy, bispecific antibodies, combination strategies with chemotherapy or targeted therapy, cancer vaccines, and adoptive cell therapy. Despite some progress, the overall response to immunotherapy remains suboptimal, and future strategies need to focus on deciphering context-specific resistance mechanisms and enhancing the tumor-specific immune response. Full article

(This article belongs to the Section Molecular Oncology)

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11 pages, 818 KB

Open AccessArticle

Clinical Manifestations and Genetic Profile of Chinese Patients with NK-Cell Large Granular Lymphocytic Leukemia—A Single-Center Retrospective Analysis

by Zhe Zhuang, Huiying Zhu, Chao Chen, Yiao Di, Zhangyuting He, Wei Zhang, Daobin Zhou and Yan Zhang

Int. J. Mol. Sci. 2026, 27(14), 6227; https://doi.org/10.3390/ijms27146227 - 13 Jul 2026

Abstract

Natural killer cell large granular lymphocytic leukemia (NK-LGLL) is a rare and heterogenous lymphoproliferative disorder. This study retrospectively evaluated 35 consecutive Chinese patients (median age 58 years) to evaluate their unique clinical–biological profiles and treatment responses. Our Chinese population exhibited a distinct comorbidity [...] Read more.

Natural killer cell large granular lymphocytic leukemia (NK-LGLL) is a rare and heterogenous lymphoproliferative disorder. This study retrospectively evaluated 35 consecutive Chinese patients (median age 58 years) to evaluate their unique clinical–biological profiles and treatment responses. Our Chinese population exhibited a distinct comorbidity spectrum, characterized by a lower prevalence of concurrent arthritis (2.9%) and secondary malignancies, compared with Western cohorts. At diagnosis, 31.4% of the cohort had neutropenia, 42.9% had anemia, and 31.4% had thrombocytopenia. The median large granular lymphocyte count was 3.9 × 10⁹/L (range 0.11–114.8 × 10⁹/L; IQR 1.9 × 10⁹/L, 5.9 × 10⁹/L). Immunophenotyping consistently identified as a CD3 CD56+ clone. Notably, genomic profiling via NGS revealed a STAT3 mutation rate of 14.3%. Regarding therapeutic efficacy, frontline immunosuppressive therapy with cyclophosphamide or cyclosporine was associated with favorable clinical responses (best overall response, complete remission rate 66.7% for both). Additionally, sirolimus emerged as a potentially highly effective salvage option, yielding an overall response rate of 85.7% (95%CI 42.1–99.6%) and complete remission rate of 57.1%. With an estimated 3-year overall survival rate of 85.6% (95%CI 73.3%, 99.8%), our findings suggest a generally indolent clinical course of NK-LGLL in this Chinese cohort and highlight the potential of mTOR inhibition in refractory cases, warranting further prospective investigation. Full article

(This article belongs to the Section Molecular Immunology)

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20 pages, 7906 KB

Open AccessArticle

Targeting Phosphatidylserine Synthesis for Tumor Cell Suppression in Esophageal Squamous Cell Carcinoma and Glioblastoma

by Yixuan Hu, Yaqi Cui, Zeqiong Xu, Duo Wu and Xiaochun Yu

Int. J. Mol. Sci. 2026, 27(14), 6226; https://doi.org/10.3390/ijms27146226 - 13 Jul 2026

Abstract

While altered lipid metabolism is a hallmark of cancer, specific phospholipid dependencies remain poorly defined. Here, we identify phosphatidylserine synthase 1 (PTDSS1) as a targetable metabolic vulnerability in esophageal squamous cell carcinoma (ESCC) and glioblastoma (GBM). Pharmacological inhibition of PTDSS1 selectively and potently [...] Read more.

While altered lipid metabolism is a hallmark of cancer, specific phospholipid dependencies remain poorly defined. Here, we identify phosphatidylserine synthase 1 (PTDSS1) as a targetable metabolic vulnerability in esophageal squamous cell carcinoma (ESCC) and glioblastoma (GBM). Pharmacological inhibition of PTDSS1 selectively and potently suppresses tumor growth both in vitro and in vivo. Mechanistically, PTDSS1 blockade triggers a rapid collapse of cellular phosphatidylserine (PS) and phosphatidylethanolamine (PE) pools, fundamentally disrupting endoplasmic reticulum (ER) homeostasis. This targeted lipid depletion activates a PERK-mediated autophagic response that ultimately yields to apoptosis. Clinically, pan-cancer transcriptomic analysis links elevated PTDSS1 expression to reduced overall survival across diverse malignancies. Collectively, we establish PTDSS1 as an essential maintainer of ER integrity and highlight PS biosynthesis as a viable therapeutic target for exploiting tumor-specific metabolic dependencies. Full article

(This article belongs to the Section Molecular Oncology)

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17 pages, 8657 KB

Open AccessArticle

Soluble Immune Checkpoints as Prognostic Biomarkers in Small Cell Lung Cancer Patients Treated with Chemotherapy and Anti-PD-L1

by Albert Guinart-Cuadra, Aida Piedra, Sergio Martínez-Recio, Maria Mulet, Carlos Zamora, Rubén Osuna-Gómez, Elisabet Cantó, Maria Angels Ortiz, Andrés Barba, Judit Sanz-Beltran, Jorgina Serra-López, Luís Paz-Ares, Edurne Arriola, Alberto Luis Moreno, Rosario Garcia-Campelo, Cristina Martí-Blanco, Dolores Isla, Ángel Callejo, Margarita Majem and Silvia Vidal

Int. J. Mol. Sci. 2026, 27(14), 6225; https://doi.org/10.3390/ijms27146225 - 13 Jul 2026

Abstract

Soluble immune checkpoints (sICs) have emerged as potential biomarkers in various cancers. However, their role in extensive-stage small cell lung cancer (ES-SCLC), an aggressive tumor type with limited therapeutic options and poor prognosis, remains poorly characterized. We analyzed 14 circulating sICs, leukocyte–platelet (PLT) [...] Read more.

Soluble immune checkpoints (sICs) have emerged as potential biomarkers in various cancers. However, their role in extensive-stage small cell lung cancer (ES-SCLC), an aggressive tumor type with limited therapeutic options and poor prognosis, remains poorly characterized. We analyzed 14 circulating sICs, leukocyte–platelet (PLT) complexes, and PD-L1 surface expression in ES-SCLC patients treated with chemoimmunotherapy (n = 41) prior to treatment and healthy donors (HD, n = 10). We assessed their associations with clinical and demographic factors and performed survival analyses using Kaplan–Meier and log-rank tests. Levels of soluble (s) PD-L1, PD-L2, BTLA, HVEM, TIM-3, and CD27 were higher in plasma from ES-SCLC patients compared to those from HD (p < 0.05). Patients with liver metastases exhibited higher sIC levels, and a multivariate model demonstrated discriminative power. Network analyses revealed distinct patterns of immune dysregulation between ES-SCLC and HD. We observed correlations among sICs and leukocyte–PLT complexes and leukocyte PD-L1 expression, indicating links between systemic immune status and tumor–immune interactions. Furthermore, increased concentrations of sTIM-3, sCD27, sHVEM and sPD-L1 were associated with a poor prognosis. Overall, sICs reflect relevant clinical, prognostic, and immunological features in ES-SCLC. Their plasma measurement could aid in non-invasive patient stratification regarding liver metastases and survival risk. Full article

(This article belongs to the Special Issue Molecular Biomarkers in Cancers: Advances and Challenges, 2nd Edition)

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16 pages, 1805 KB

Open AccessArticle

Structural Modifications of Hybrid O-Alkylsulfonyl-β-(Benzimidazol-1-yl)propioamidoximes as a Pathway to the Antimicrobial, Antifungal and Antidiabetic Drugs

by Lyudmila Kayukova, Anna Vologzhanina, Ekaterina Dubasova, Roza Seidakhmetova, Azamat Yerlanuly, Aruzhan Sartoyeva and Aigul Malmakova

Int. J. Mol. Sci. 2026, 27(14), 6224; https://doi.org/10.3390/ijms27146224 - 12 Jul 2026

Abstract

The continuous search for new chemical structures more active and less toxic than those currently in practice is justified on the basis of the use of proven pharmacophoric building blocks (benzimidazole heterocycle, sulfonyl group and amidoxime framework in our case). The aim of [...] Read more.

The continuous search for new chemical structures more active and less toxic than those currently in practice is justified on the basis of the use of proven pharmacophoric building blocks (benzimidazole heterocycle, sulfonyl group and amidoxime framework in our case). The aim of the work was to synthesize new O-alkylsulfonyl-β-(benzimidazol-1-yl)propioamidoximes and to test them for in vitro biological activities to identify potentially effective agents. Novel O-alkylsulfonylamidoximes were synthesized in hydrochloride and base forms by the reaction of β-(benzimidazol-1-yl)propioamidoxime with alkylsulfonyl chlorides AlkSO₂Cl (Alk = CH₃, n-C₃H₇, i-C₃H₇, n-C₄H₉) in a mixture of water:acetone in hydrochloride and base forms. Hydrochlorides and bases of the O-alkylsulfonyl-β-(benzimidazol-1-yl)propioamidoximes were obtained in moderate to high yields. The structures of the synthesized compounds were established by physicochemical and spectral (elemental analysis, FT-IR, NMR and X-ray diffraction) methods. The obtained derivatives were tested for antimicrobial, antifungal and antidiabetic activity. Biological screening found effective samples of amidoximes with antimicrobial and antifungal activities that were near or exceeded the activity of the reference drugs gentamicin and nystatin; in addition, two samples with antidiabetic activity higher than acarbose were found. The results of the present study open new possibilities for the novel β-aminopropioamidoxime class as active antimicrobial and antifungal agents, as well as antidiabetic ones. Full article

(This article belongs to the Special Issue Advances in Organic Synthesis in Drug Discovery)

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16 pages, 1144 KB

Open AccessCase Report

Genotype-Driven Diagnosis Enables Targeted Pharmacological Treatment in Brunner Syndrome: A Novel Splice-Site MAOA Variant and Case-Based Review

by Elisa Gravagno, Melissa Bellini, Enrico Ambrosini, Anita Luberto, Sabrina Busciglio, Giulia Vitetta, Ilenia Rita Cannizzaro, Antonietta Taiani, Valeria Barili, Antonio Percesepe, Vera Uliana and Davide Martorana

Int. J. Mol. Sci. 2026, 27(14), 6223; https://doi.org/10.3390/ijms27146223 - 12 Jul 2026

Abstract

Brunner syndrome is a rare X-linked neurodevelopmental disorder caused by loss-of-function (LOF) variants in the monoamine oxidase A gene (MAOA), which encodes monoamine oxidase A, a key enzyme involved in the degradation of monoamine neurotransmitters such as serotonin, norepinephrine, and epinephrine. [...] Read more.

Brunner syndrome is a rare X-linked neurodevelopmental disorder caused by loss-of-function (LOF) variants in the monoamine oxidase A gene (MAOA), which encodes monoamine oxidase A, a key enzyme involved in the degradation of monoamine neurotransmitters such as serotonin, norepinephrine, and epinephrine. Impaired MAOA activity leads to abnormal monoamine accumulation and disruption of monoaminergic signalling, resulting in intellectual disability and behavioural dysregulation. Here, we systematically summarize the molecular landscape and report a genotype-driven diagnosis of MAOA deficiency in a patient presenting with intellectual disability and no reported family history. Clinical exome sequencing (cES) identified a novel splice-site variant in the MAOA gene that had not been detected by first-line diagnostic approaches. Functional analysis of patient-derived mRNA demonstrated intron 8 retention leading to a premature stop codon, consistent with a LOF mechanism. Based on the molecular diagnosis, the patient received treatment with serotonin antagonist and reuptake inhibitor (SARI) class medication, which was associated with improvement in social behaviour and sleep disturbances. Notably, to the best of our knowledge, this represents the first reported use of SARI therapy in MAOA deficiency. Although SARI therapy in this condition remains off-label, this observation provides preliminary evidence suggesting a potential therapeutic benefit. Our findings expand the mutational spectrum of the MAOA gene and highlight the importance of molecular diagnosis driving personalized management in rare neurogenetic disorders. Full article

(This article belongs to the Special Issue Molecular Mechanisms Underlying the Pathogenesis of Genetic Diseases)

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