Search Results (1,190)

Hepatocellular carcinoma (HCC) remains a major global health burden, characterized by late diagnosis, limited therapeutic options, and high mortality rates. Conventional diagnostic tools such as serum α-fetoprotein testing and imaging lack sufficient sensitivity for early detection. In recent years, liquid biopsy has emerged as a minimally invasive approach that enables real-time molecular profiling of tumors through the analysis of circulating biomarkers such as nucleic acids, proteins, and extracellular vesicles. Recent advances have underscored exosomes—nano-sized extracellular vesicles (EVs) secreted by nearly all cell types—as pivotal mediators of intercellular communication and dynamic carriers of tumor-derived molecular information, offering exciting prospects for early cancer detection and personalized therapy. In HCC, EV microRNAs (miRNAs) participate in multiple oncogenic processes, including proliferation, angiogenesis, epithelial–mesenchymal transition, and immune modulation. Specific EV-associated miRNAs, such as miR-21, miR-122, miR-224, and miR-221, show distinctive expression profiles in HCC and correlate with tumor stage, metastasis, and patient prognosis. Moreover, panels of circulating EV-associated miRNAs demonstrate superior diagnostic accuracy compared with traditional biomarkers, underscoring their potential as non-invasive tools for early detection and disease monitoring. Their inherent stability in biofluids and resistance to enzymatic degradation further support their application in liquid biopsy approaches. Despite promising results, continued research is essential to validate EV-associated miRNA signatures and to integrate these “silent messengers” into routine clinical practice for precision management of hepatocellular carcinoma. Full article

The global expansion of highly pathogenic avian influenza A (H5N1), particularly the clade 2.3.4.4b lineage, has renewed urgent concerns about its pandemic potential in the context of its ongoing panzootic expansion and increasing cross-species transmission. Despite decades of preparedness initiatives, critical technological and structural gaps persist, especially in low- and middle-income countries (LMICs), where both vaccine access and sustainable manufacturing capacity remain limited. In this perspective, we examine key lessons from past influenza pandemics and global preparedness strategies, including the Global Action Plan for Influenza Vaccines, highlighting persistent challenges related to sustainable manufacturing capacity and equitable vaccine access. Additionally, we examine the potential of messenger RNA (mRNA) vaccine platforms to address these limitations, given their rapid design, scalable manufacturing, and adaptability to emerging pathogens. Moreover, we examine the role of neuraminidase (NA) as a complementary antigen capable of broadening immune protection and reducing viral transmission. Finally, we describe recent advances in Latin America, focusing on Argentina’s participation in the mRNA Technology Transfer Programme co-led by the World Health Organization (WHO) and the Medicines Patent Pool (MPP), as a model for strengthening regional manufacturing capacity and contributing to global pandemic preparedness. Together, these elements indicate that effective H5N1 pandemic preparedness will require the integration of improved antigen design, flexible mRNA platforms, and sustainable regional manufacturing systems aligned with global procurement strategies. Full article

The northern pike (Esox lucius) is an economically important cold-water fish species in northern China. It exhibits pronounced sexual dimorphism, yet the molecular mechanism underlying its sex differentiation remains unclear, which hinders the development of aquaculture. Whole-transcriptome sequencing is a powerful approach for screening sex-related genes; however, no such study has been reported for this species to date. In this study, gonadal tissues from three female and three male E. lucius were collected for whole-transcriptome sequencing. A total of 14,941 differentially expressed messengerRNAs, 119 differentially expressed microRNAs, 229 differentially expressed circularRNAs, and 2055 differentially expressed long non-codingRNAs were identified. Functional enrichment analysis revealed that the differentially expressed genes were significantly enriched in pathways closely associated with sex differentiation, such as steroid hormone biosynthesis and oocyte meiosis. Several key sex-biased genes were identified, including female-biased genes (FANCL, DDX5, SRSF5B) and male-biased genes (STAR, FDX1B, ITGA2B). Furthermore, a competing endogenous RNA (ceRNA) regulatory network involving dre-miR-107b was constructed, which may represent a candidate for further investigation into sex differentiation in E. lucius. This study provides the first comprehensive whole-transcriptome dataset of female and male gonads in E. lucius, identifies key sex-biased genes and core pathways involved in its sex differentiation, and thereby identifies the dre-miR-107b-centered ceRNA network and key sex-biased genes (FANCL, DDX5, SRSF5B, STAR, FDX1B, ITGA2B) as core molecular players in sex differentiation of this species. Full article

Tuberculosis (TB) remains a major global public health burden. This study aimed to identify differentially expressed messenger RNAs (mRNAs) and circulating microRNAs (miRNAs) associated with TB and to validate their potential roles in the disease. We performed RNA sequencing (RNA-Seq) on peripheral blood samples from 10 patients with active pulmonary TB and 10 healthy controls, using peripheral blood mononuclear cells (PBMCs) for mRNA sequencing and plasma for miRNA sequencing. Given the exploratory nature of the plasma miRNA data and the limitations of the U6 normalization method, the results for circulating miRNAs will need to be validated using alternative methods in subsequent experiments. A total of 1323 differentially expressed mRNAs and 49 differentially expressed miRNAs were identified. Functional annotation of differentially expressed genes was conducted using the Database for Annotation, Visualization and Integrated Discovery (DAVID), followed by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, which revealed two TB-associated pathways: “MicroRNAs in cancer” and “Small cell lung cancer.” Two key mRNAs—tumor protein p53 (TP53) and forkhead box protein P1 (FOXP1)—and one key miRNA (hsa-miR-29b-3p) were identified as potential core regulatory factors. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) validation confirmed that the expression patterns of these candidate molecules were consistent with the RNA-Seq results. Three potential candidate molecules associated with TB were ultimately identified, although their disease specificity remains to be determined. Full article

Chimeric antigen receptor T (CAR-T) cell therapy has transformed the treatment of hematologic malignancies, yet its broader application, particularly in solid tumors, remains constrained by high cost, labor-intensive manufacturing, limited production capacity, and variable clinical performance, as well as barriers such as poor trafficking, antigen heterogeneity, and an immunosuppressive tumor microenvironment. In vivo CAR-T cell engineering, in which CAR-T cells are generated directly within the patient, offers a paradigm shift by eliminating the need for ex vivo cell processing and complex logistical infrastructure. Among emerging approaches, messenger RNA (mRNA)-loaded lipid nanoparticles (LNPs) have emerged as a promising and clinically tractable platform for in vivo CAR-T cell generation, enabling direct reprogramming of T lymphocytes within the patient and thereby circumventing the need for leukapheresis, viral vector production, and prolonged ex vivo culture, effectively transforming the patient into their own cell therapy factory. This review synthesizes advances in mRNA-LNP-mediated in vivo CAR-T cell generation, encompassing ionizable lipid chemistry and emerging T cell-targeted delivery strategies, including surface functionalization approaches. We discuss the implications of transient CAR expression for immune activation, safety, and therapeutic durability, alongside CAR design optimization through co-stimulatory domains and safety switches. Preclinical evidence from murine tumor models and non-human primates is integrated with current regulatory considerations, and key barriers to clinical translation are highlighted. Collectively, progress in nucleic acid delivery, synthetic immunology, and precision medicine positions in vivo mRNA-CAR-T therapy as a promising modality for oncology and beyond. Full article

Placenta-derived extracellular vesicles (EVs), particularly exosomes, serve as key mediators that influence metabolic programming in offspring under adverse early nutritional conditions, such as maternal obesity or gestational diabetes. They respond to maternal nutritional disturbances—such as obesity or gestational diabetes—by altering the composition of the miRNAs and proteins they carry. Evidence from in vivo and in vitro studies suggests that these modified EVs influence offspring metabolic programming through multiple putative pathways: regulating fetal pancreatic β-cell development and function, modulating lipogenesis via PPARγ signaling, affecting placental angiogenesis, and promoting inflammation and epigenetic alterations. By transmitting maternal environmental signals to the fetus, placental EVs are hypothesized to contribute to long-term metabolic phenotypes and disease susceptibility. This review critically examines the current evidence positioning placental EVs as key messengers in maternal–fetal communication, evaluates the strength of evidence supporting their role in shaping offspring metabolic health, identifies major knowledge gaps (e.g., limited direct evidence in human offspring, lack of standardized isolation methods), and suggests their potential as early intervention biomarkers or therapeutic targets for preventing metabolic disorders in offspring. We also highlight the need for prospective cohort studies and mechanistic validation in appropriate animal models to establish causality. Full article

Lipid nanoparticles (LNPs) have emerged as popular nucleic acid delivery systems, yet the dynamic mechanisms related to their self-assembly and structural maturation remain insufficiently understood due to the limitations of traditional offline characterization tools. This study establishes a time-resolved (TR) in situ small-angle X-ray scattering (SAXS) methodology to monitor the structural evolution of LNPs during microfluidic formulation and subsequent maturation. By integrating a dual-channel microfluidic mixing system with a SAXS measurement platform, we successfully captured the real-time scattering profiles of both empty and messenger RNA-loaded nanoparticles (mRNA-LNPs). The results demonstrate distinct assembly pathways for empty-LNPs and those encapsulated with mRNA. The empty-LNPs undergo a gradual transition toward periodic nanostructures, whereas mRNA-LNPs exhibit rapid complexation into stable subunits followed by hierarchical assembly. Furthermore, the platform effectively tracked nanoscale structural rearrangements during a microfluidic dilution process, revealed by subtle shifts in scattering peaks and internal periodicity. Overall, this time-resolved approach provides a robust experimental framework for capturing transient intermediate states, offering a valuable tool to elucidate molecular assembly mechanisms and facilitate the rational design of next-generation nanomedicines. Full article

Messenger RNA (mRNA) therapeutics are redefining treatment approaches in vaccines, cancer immunotherapy, protein replacement, and gene editing. Lipid nanoparticles have enabled early clinical successes, but they can be limited by liver-dominant biodistribution, long-term storage stability, and systemic tolerability. Polymeric delivery systems offer a versatile alternative, with tunable physicochemical properties enabling precise control over mRNA complexation, protection, release, and targeting. This review examines recent progress across polyethyleneimine derivatives, poly(β-amino ester)s, poly(amino acid)s, polyesters, dendrimers, charge-altering releasable transporters, and lipid-polymer hybrids. We highlight strategies such as structural modification, stimuli-responsive designs, and high-throughput polymer screening that enhance stability, reduce cytotoxicity, and enable organ- or cell-specific delivery. Addressing challenges in immunogenicity, biodistribution, and manufacturing scalability will be pivotal to translating these innovations into safe and effective mRNA therapeutics. Full article

Zoonotic tuberculosis (zoonotic TB) caused by Mycobacterium bovis (M. bovis) accounts for up to 10% of human tuberculosis cases in some regions, but the underlying pathogenic mechanisms remain incompletely understood, especially those involved in cellular pyroptosis. This study aimed to explore the regulatory roles of non-coding RNA (ncRNA) in the pyroptosis of human monocytic THP-1 cells induced by M. bovis infection. An in vitro pyroptosis model was established by infecting THP-1 cells with M. bovis, followed by whole-transcriptome sequencing to identify differentially expressed messenger RNA (mRNA), long non-coding RNA (lncRNA), microRNA (miRNA), and circular RNA (circRNA). Bioinformatics analysis was performed to construct an lncRNA–miRNA–mRNA regulatory network associated with infection-induced pyroptosis; in addition, overexpression, knockdown, and dual-luciferase reporter assays and quantitative PCR were conducted to verify the interactions and functions of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), miR-20b-5p, and purinergic receptor P2X7 (P2RX7). Transcriptome analysis detected 741 mRNAs, 1049 lncRNAs, 25 circRNAs, and 40 miRNAs with significant differential expression in infected THP-1 cells. Specifically, MALAT1 and P2RX7 were upregulated, while miR-20b-5p was downregulated after infection. Knockdown of MALAT1 or P2RX7 and overexpression of miR-20b-5p relieved M. bovis-induced pyroptosis in THP-1 cells. Mechanistically, MALAT1 targeted miR-20b-5p, which directly targeted P2RX7, and overexpression of miR-20b-5p partially reversed P2RX7 upregulation mediated by MALAT1 overexpression. This study provides a transcriptomic characterization of M. bovis-induced pyroptosis in THP-1 cells and supports the MALAT1–miR-20b-5p–P2RX7 axis as a potential regulatory mechanism involved in this process, offering initial molecular insights into the pathogenesis of zoonotic TB. Full article

Messenger RNA (mRNA) vaccines are emerging as promising tools capable of reshaping how cancer interacts with the immune system and responds to immunotherapy. These vaccines not only act as platforms for antigen delivery but can also influence the tumor microenvironment (TME), fostering a shift from immunologically “cold’’ conditions toward “hotter’’ and treatment-responsive states. In melanoma, this capability has been found to enhance the efficacy of the immune checkpoint inhibitors (ICIs), as mRNA-based priming can provide the robust antitumor activation needed for more effective checkpoint blockade. Early clinical studies with personalized or off-the-shelf vaccines showed benefits in patients with high-risk resected melanoma or refractory to PD-1 inhibition. Combining mRNA vaccines with ICIs, along with other immunomodulatory strategies, may be helpful to overcome resistance arising from the TME and achieve more durable clinical benefits. Besides these advances, computational and in silico modeling are providing new insights into how mRNA vaccines modulate the TME, helping to identify factors such as antigen-presenting cell (APC) density, CD8⁺ T-cell infiltration, and macrophage polarization that may predict treatment success and guide personalized strategies. Together, these developments indicate that combining mRNA vaccination with ICIs, supported by computational tools, may improve clinical outcomes in melanoma and, potentially, in selected tumor types with favorable immunological features, although important biological constraints limit direct extrapolation to less immunogenic malignancies. Full article

Segmented and non-segmented negative-strand RNA viruses share the same general pathway for genome transcription, which generates messenger RNA, and genome replication which duplicates the viral RNA. These processes are performed by the viral polymerase and necessitate the viral RNA to be coated by a non-covalent polymer of nucleoproteins known as nucleocapsid. The non-segmented negative-strand RNA viruses (nsNSVs) have rigid nucleocapsids covering the entire tightly bound genome and require a phosphoprotein cofactor for proper replication and transcription by the polymerase, while the segmented negative-strand RNA viruses (sNSVs) have very flexible nucleocapsids with only few nucleotides tightly bound to each nucleoprotein, and their viral RNA genome ends are directly bound to the polymerase. We discuss here how the differences in RNA binding are likely to be crucial for proper replication and transcription in both nsNSVs and sNSVs. Full article

Triple-negative breast cancer is defined by the absence of druggable receptor targets and by a biologically dynamic phenotype that renders static, single-timepoint biomarker strategies fundamentally inadequate. Current predictive markers, including PD-L1 expression, tumor mutational burden, and genomic profiling, fail to capture the therapy-induced transcriptional reprogramming, spatial heterogeneity, and drug-tolerant persister states that drive resistance and relapse. In this review, we argue that RNA, particularly non-coding RNA (ncRNA), represents a complementary and state-aware platform for biomarker development in TNBC, capable of capturing transcriptional adaptation, regulatory threshold dynamics, and cell state transitions that static genomic markers cannot fully detect. Unlike messenger RNAs, which reflect active transcriptional programs, long non-coding RNAs and circular RNAs modulate the stability of state transitions and are specifically induced under conditions of therapeutic stress, immune exclusion, and drug tolerance, which are properties that make them suitable as potential early and sensitive indicators of adaptive reprogramming. We review the biological rationale for RNA as a state-aware readout across five dimensions: tumor plasticity, immune context, stress response, therapy adaptation, and microenvironment composition. An examination is conducted regarding how spatial transcriptomics can map RNA-defined resistant niches within TNBC, how serial liquid biopsy RNA measurements, including extracellular vesicle RNA and circulating tumor RNA, enable temporal monitoring of transcriptional state shifts before radiologic progression, and what analytical and clinical standards deployable RNA assays must meet. Finally, a state-guided adaptive management framework is proposed in which RNA signatures function as iteratively updated measurement layers informing therapy selection, on-treatment monitoring, and early resistance detection. This review outlines trial design models and defines the validation standards required before RNA-guided adaptation can enter clinical practice. Full article

mRNA vaccines are a relevant approach in cancer immunotherapy, using messenger RNA to induce immune responses against tumor-associated antigens. In this review, BNT111 and BNT122 are compared as representative off-the-shelf and personalized models. BNT111 is a fixed mRNA vaccine that has demonstrated significant antitumor efficacy against shared melanoma antigens, particularly when combined with immune checkpoint inhibitors. It allows a standardized production via in vitro transcription (IVT) in a cell-free system. Conversely, BNT122 is a personalized vaccine designed to match an individual’s tumor mutations by targeting patient-specific neoantigens to elicit more robust immune responses. It has significant suitability in the adjuvant setting to target minimal residual disease. Despite favorable safety and immunogenicity, the effectiveness of these vaccines is influenced by various factors, including tumor heterogeneity, differences in antigen expression, off-target effects on mRNA-LNP distribution, molecular instability, and complex manufacturing constraints. Neither approach can be directly considered as the definitive optimal vaccine. A comprehensive analysis of their strengths and limitations is vital for a balanced and objective future research direction. Collectively, this emphasizes the need for further improvements in vaccine design and strategies, prioritizing high-quality, safe, and accessible treatments for every cancer-based patient and ensuring their successful integration into healthcare. Full article

Triple-negative breast cancer is an aggressive and heterogeneous type of invasive breast cancer (BC) in which the cancer cells lack estrogen and progesterone receptors, as well as expression of the human epidermal growth factor 2 protein. This cancer tends to grow and spread faster than other BC subtypes, and is associated with a poor prognosis due to early visceral and neurological recurrences. Multidisciplinary management includes surgery, chemotherapy, radiation therapy, and immunotherapy with targeted immune checkpoint inhibitors (ICIs). The introduction of ICIs has improved outcomes in patients with TNBC, particularly in the metastatic and neoadjuvant settings. Despite these advances, a significant proportion of patients either do not respond to treatment or develop resistance to it. TNBC mortality remains high, underscoring the urgent need to identify novel prognostic and predictive biomarkers to overcome resistance to immunotherapy. Following a brief overview of the clinical features and established biomarkers of TNBC, the current review focuses on immune checkpoint proteins (ICPs) beyond PD-1 and PD-L1, and on the potential use of soluble ICPs rather than the well-established membrane-bound assays. These soluble ICPs are produced through the alternative splicing of messenger (m)RNA or the cleavage/shedding of membrane-bound proteins. This is followed by an overview of current treatment and novel predictive targets in TNBC. Additionally, the involvement of the B- and T-lymphocyte attenuator (BTLA)/herpes virus entry mediator (HVEM)/CD160 pathway and its role in the pathogenesis of BC and TNBC are reviewed, highlighting the potential use of BTLA and HVEM as biomarkers. Full article

Fragile X Syndrome (FXS) is an inherited cause of intellectual disability and autism, arising from silencing of the Fmr1 gene and loss of Fragile X Messenger Ribonucleoprotein 1 (FMRP). FMRP is an RNA-binding protein critically involved in neurodevelopmental processes, including neurogenesis. We examined the proliferation and maturation of adult-born dentate granule cells (abDGCs) and glial populations in Fmr1 knockout (KO) and wild-type (WT) mice at 4, 12, and 24 weeks of age under control and early-life stress (ELS) conditions. Based on prior findings, we hypothesized that KO mice would exhibit increased neurogenesis and atypical responses to ELS compared with WT mice. Using immunohistochemistry, we quantified multiple stages of neurogenesis in the dentate gyrus, including proliferating (Ki67+), immature (doublecortin [DCX]+), and apoptotic (cleaved caspase-3 [CC3]+) cells. We also assessed glia using Iba1 (microglia) and GFAP (astrocytes) immunoreactivity. KO mice displayed significantly increased Ki67+ proliferating and reduced CC3+ apoptotic cells across ages, accompanied by increased Iba1+ and GFAP+ glial densities. However, KO mice exhibited fewer DCX+ neuroblasts at later time points. When reared in ELS conditions, KO mice show blunted or no changes in neurogenesis and glial populations relative to WT mice reared in ELS conditions or KO mice in control conditions. These results indicate that FMRP loss disrupts hippocampal neurogenesis by increasing cell proliferation while limiting neuronal maturation and expanding glial populations. Moreover, the absence of neurogenic and glial responses to ELS in KO mice highlights a gene–environment interaction that may influence FXS-related neuropathology by limiting the adaptive capacity of the hippocampal neurogenic niche. Full article