Search Results (588)

Background: Ligamentum flavum hypertrophy (LFH) is a major structural contributor to lumbar spinal stenosis and is characterized by extracellular matrix (ECM) remodeling with an increasingly recognized immune-associated component. However, the regulatory architecture linking disease-associated microRNA (miRNA) dysregulation to LFH transcriptomic remodeling remains incompletely defined. Methods: Public Gene Expression Omnibus datasets were analyzed using an integrated mRNA–miRNA transcriptomic framework. Single-cell RNA sequencing (GSE294458) was used to characterize the cellular landscape of hypertrophic and non-hypertrophic ligamentum flavum, whereas bulk transcriptomic analysis (GSE113212) identified LFH-associated differentially expressed genes. Differentially expressed miRNAs from an ossified ligamentum flavum dataset (GSE106256) were integrated with LFH-associated mRNA profiles through inverse miRNA–mRNA regulatory filtering. Functional enrichment, STRING protein–protein interaction (PPI) analysis, cytoHubba hub gene prioritization, LASSO regression, ROC analysis, remodeling-signature scoring, DGIdb, ChEA, and database-supported miRNA–target annotation were subsequently performed. Results: Single-cell analysis supported fibroblast-, myofibroblast-, and ECM-associated remodeling in hypertrophic ligamentum flavum. Bulk analysis identified nine significant differentially expressed genes, and integration with 33 dysregulated miRNAs generated 651 inverse-regulated core genes. Enrichment analyses highlighted ECM organization, proteoglycan/glycosaminoglycan (GAG) biology, immune cell differentiation, antigen presentation, and NF-κB/Wnt-related pathways. The STRING network included 650 nodes and 547 edges, with significant PPI enrichment (p = 2.04 × 10⁻¹⁴). LASSO prioritized PABPC1 and RPL4 as exploratory candidate hub features showing apparent discovery-cohort discrimination (AUC = 1.00); supplementary uncertainty, internal-stability, and separation-aware sensitivity analyses supported interpretation of this finding as a discovery-cohort signal rather than as independent diagnostic validation. Remodeling-signature analyses showed increased ECM fibrosis and proteoglycan/GAG scores, with inverse associations involving PABPC1 and RPL4. Multi-layer prioritization identified hsa-miR-708-5p as the leading candidate, followed by hsa-miR-23b-3p, hsa-miR-191-5p, hsa-miR-181a-5p, and hsa-miR-653-5p. Conclusions: This integrated mRNA–miRNA transcriptomic analysis delineated a coordinated fibrotic–immune remodeling landscape in LFH and prioritized experimentally testable miRNA candidates linked to network-central regulatory pathways. Full article

Background: Triple-negative breast cancer (TNBC) is an aggressive subtype with persistent disparities in stage at diagnosis. While transportation-related exposures are increasingly recognized as environmental health risks, the biological mechanisms linking these exposures to cancer progression remain unclear. This study evaluated whether tumor microRNAs (miRNAs), which regulate gene expression and tumor behavior, mediate the association between transportation burden and TNBC stage at diagnosis. Methods: We analyzed 434 TNBC cases from the Louisiana Tumor Registry (2009–2019). The transportation burden from the 2022 Environmental Justice Index reflects the residential proximity to high-volume roads, railways, and airports. miRNA expression was measured via high-throughput sequencing and normalized using the trimmed mean of M-values method. Three-phase analysis (screening, individual, and multiple mediation) was performed. KEGG pathway enrichment analysis assessed downstream biological pathways. Results: After adjusting for age, race, body mass index, marital status, primary payer, and concentrated disadvantage index (CDI), every 0.10 (10-percentile) increase in transportation burden rank was significantly associated with a 9% increase in the odds of late-stage TNBC diagnosis (Adjusted OR = 1.09, 95% CI: 1.01–1.17, p = 0.021). Five miRNAs significantly mediated this relationship: downregulated hsa-let-7c-5p, hsa-let-7b-5p, hsa-miR-30a-3p, and hsa-miR-92a-3p, and upregulated hsa-miR-151a-3p. Joint mediation analysis demonstrated complete mediation (indirect effect = 0.324; p = 0.027). hsa-let-7c-5p was the primary independent mediator. The enriched pathways included MAPK, PI3K-Akt, Wnt, and p53 signaling. Conclusions: These findings identified molecular pathways linking transportation-related environmental exposures to TNBC progression through dysregulation of miRNAs. By integrating environmental exposure assessment with tumor biology, this study advances our understanding of how the built environment contributes to cancer disparities. Full article

Background and Objectives: The clinical differentiation of peripheral involvement within the spondyloarthritis (SpA) spectrum remains a significant challenge. Identifying patients at the stage of psoriasis without clinical arthritis (PsO), before the onset of overt arthritis, is crucial for early disease management. MicroRNAs (miRNAs) have emerged as potential epigenetic regulators in inflammatory rheumatic diseases. This study aimed to identify circulating miRNA profiles that serve as discriminative biomarkers between PsO and peripheral SpA (p-SpA). Materials and Methods: This exploratory study was conducted in two phases. In the preliminary discovery phase, plasma miRNA expression was analyzed using high-throughput microarrays in patients with p-SpA (modeled by peripheral psoriatic arthritis, n = 6), PsO (psoriasis without clinical arthritis, n = 3), and osteoarthritis (n = 3). In the validation phase, candidate miRNAs were verified using TaqMan-based qPCR in an independent cohort (n = 30) of various SpA phenotypes, categorized into those with peripheral arthritis (SpA-A) and those without (SpA-N). Results: The preliminary discovery phase identified altered levels of hsa-miR-140-5p, hsa-miR-192-5p, and hsa-miR-146a-5p among the groups; however, due to the small sample size, these initial findings required strict downstream verification. Functional enrichment analysis revealed that these candidate miRNAs primarily targeted the NF-κB signaling pathway (hsa04064) and Toll-like receptor (TLR) signaling pathway (hsa04620). The validation cohort confirmed that these three miRNAs could reliably differentiate SpA-A from SpA-N patients. Furthermore, bioinformatic mapping predicted that downstream targets, including TRAF6, IRAK1, and CXCL2, may be associated with these clinical phenotypes, serving as hypothesis-generating observations for future studies. Conclusions: Our findings suggest that specific plasma miRNA profiles are associated with the inflammatory pathways driving peripheral involvement in the SpA spectrum. These miRNAs represent potential biomarkers associated with peripheral arthritis in the SpA spectrum. While they offer new molecular insights into disease pathogenesis, their predictive value for identifying PsO patients at risk of developing overt arthritis requires confirmation in future longitudinal studies. Full article

Objective: Lung cancer (LC) remains a leading cause of cancer mortality worldwide. Thymoquinone (TQ), a bioactive compound derived from Nigella sativa, possesses anti-inflammatory and antioxidant properties, but its precise mechanisms concerning miRNA regulation in LC are poorly defined. This study investigates the anti-cancer potential of TQ through modulation of microRNA signaling in LC. Methods: We employed an integrated approach combining bioinformatic predictions with rigorous experimental validation in A549 lung adenocarcinoma cells and SHP-77 human small-cell lung carcinoma (SCLC) cells. Bioinformatic analyses predicted miRNA targets, and experimental techniques included dual-luciferase reporter assays, miRNA inhibition, TaqMan RT-qPCR, cell-based ELISA, and Western blotting to dissect the molecular pathway. Results: We identified the transcription factor BACH1 as a direct and novel target of hsa-miR-155-5p. TQ potently suppressed interferon-γ-induced expression of both hsa-miR-155-5p and its target, BACH1. This TQ-mediated suppression led to subsequent downregulation of the key metastasis-promoter Matrix Metalloproteinase-9 (MMP-9). Genetic inhibition of miR-155-5p or direct BACH1 inhibition phenocopied the effects of TQ, confirming the functional significance of this axis. Thus, we define a novel oncogenic signaling cascade—the hsa-miR-155-5p/BACH1/MMP-9 axis that is effectively disrupted by TQ. Conclusions: This represents the first evidence that TQ exerts its anti-cancer effects in LC through the modulation of the critical signaling cascade (hsa-miR-155-5p → BACH1 → MMP-9). Our findings establish TQ as a multi-targeted agent capable of simultaneously inhibiting miRNA-mediated oncogenic signaling and protein-level effectors. The dual therapeutic action of TQ represents a novel therapeutic strategy and underscores its potential for synergistic combination therapies. Full article

The multifaceted oncogenic role of teratocarcinoma-derived growth factor 1 (TDGF1) in colon cancer remains incompletely understood. Through integrative bioinformatic and functional analyses, we identified a novel competing endogenous RNA (ceRNA) axis wherein the long non-coding RNA OLMALINC directly sponges hsa-miR-3614-5p, leading to the derepression of TDGF1. This OLMALINC/miR-3614-5p/TDGF1 axis promoted colon cancer cell proliferation, migration, invasion, and anti-apoptosis in vitro, whereas TDGF1 knockdown significantly suppressed tumor growth in vivo. Mechanistically, TDGF1 co-activated oncogenic signaling via the Thr88-dependent Nodal/Smad2 cascade and the Glypican-1-mediated MAPK/AKT pathway. Beyond cell-autonomous effects, transcriptomic and single-cell analyses revealed that elevated TDGF1 correlates with an immunosuppressive microenvironment, characterized by reduced immune infiltration and altered LGALS9-CD44 malignant-T cell communication. Clinically, high TDGF1 expression in a tissue microarray cohort was significantly associated with advanced T stage, reduced expression of specific mismatch repair proteins (MLH1/PMS2), and poor overall survival. Collectively, this study delineates the OLMALINC/miR-3614-5p/TDGF1 regulatory circuit and establishes TDGF1 as a multifaceted driver of tumor progression, highlighting its potential as a prognostic biomarker and therapeutic target in colon cancer. Full article

MicroRNAs (miRNAs) are small non-coding RNAs that play a crucial role in the post-transcriptional regulation of gene expression. Since miRNAs modulate host immune responses, they represent promising molecular biomarkers of paratuberculosis (PTB), particularly during early or subclinical stages, when conventional diagnostic tests may lack sensitivity. In this context, faecal miRNAs could provide valuable insights into intestinal immune responses and mucosal damage associated with Mycobacterium avium subsp. paratuberculosis (MAP) infection. Although miRNAs have been extensively investigated in serum, blood, and tissues, their detection and characterization in bovine faeces remain poorly explored. The aim of this study was to evaluate the expression profiles of selected candidate faecal miRNAs in Marchigiana beef cattle naturally exposed to MAP and to assess their association with different infection phenotypes. Thirty-four cows were classified into three phenotypic groups: healthy exposed, MAP-infected, and PTB-affected based on longitudinal diagnostic records including interferon-γ assay, serological testing (ELISA), and faecal qPCR. Five candidate miRNAs were selected from previous studies and quantified in faecal samples by RT-qPCR. Four of the five selected miRNAs were consistently detected across samples. Bta-miR-92a was significantly downregulated in both MAP-infected and PTB-affected animals compared with healthy cattle, suggesting early modulation during MAP infection. Bta-miR-223 was significantly upregulated in PTB-affected animals compared with both healthy and MAP-infected groups, consistent with its established role in the regulation of intestinal inflammation. The ortholog of hsa-miR-501-5p was significantly upregulated in MAP-infected cattle, potentially reflecting early host–pathogen interactions at the intestinal mucosal level, while bta-miR-24-3p showed no significant differences among groups. Overall, these findings support the feasibility of faecal miRNA analysis as a complementary non-invasive molecular approach to support traditional diagnostic tests for PTB, especially during the early and subclinical stages of MAP infection. Full article

Immune checkpoint inhibitors (ICIs) have revolutionized the treatment of advanced non-small cell lung cancer (NSCLC). However, the limited predictive value of PD-L1 expression as a biomarker underscores the urgent need for more reliable predictors of ICI response. Interferon regulatory factor 1 (IRF1) is a transcription factor that lies downstream of interferon-γ signaling and directly regulates CD274 (PD-L1) transcription. Here, we performed a comprehensive bioinformatic analysis to identify microRNAs (miRNAs) that may be associated with IRF1 expression in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). Using data from The Cancer Genome Atlas (TCGA), we identified 20 miRNAs whose expression levels consistently and negatively correlated with IRF1 mRNA levels in both LUAD and LUSC. Among these, only hsa-miR-301b possesses conserved binding sites in the 3′UTR of IRF1 mRNA, suggesting direct post-transcriptional repression. For the remaining 19 miRNAs, we hypothesized an indirect mechanism of action. Further analysis revealed that hsa-miR-183 and hsa-miR-141 may target the transcription factor genes NFKB1 and STAT4, respectively, both of which positively correlate with IRF1 expression and are themselves associated with improved survival in ICI-treated patients. This study delineates a multi-layer miRNA regulatory network associated with IRF1 expression in NSCLC and identifies hsa-miR-301b, hsa-miR-183 and hsa-miR-141 as candidate upstream regulators of IRF1. Direct survival analysis for these miRNAs in ICI-treated cohorts was not feasible due to the lack of publicly available miRNA-seq data with treatment annotations; therefore, their clinical predictive value remains hypothetical, and experimental validation is required to assess their potential as predictors of ICI response. Full article

Human epidermal growth factor receptor 2 (HER2) remains the most recognized and clinically established molecular biomarker in gastric cancer; however, the regulatory mechanisms underlying its dysregulation are not fully understood. This study aimed to identify microRNAs associated with HER2 gene amplification, chromosome 17 centromere copy number increase (CNI), or alternative mechanisms driving HER2 protein overexpression. We analyzed 115 gastric cancer patients treated surgically at a single institution, with available material for immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), and microRNA profiling. Among 11 candidate microRNAs, four demonstrated significant associations with HER2-related alterations. hsa-miR-128-3p expression was positively associated with HER2 gene amplification, while hsa-miR-145-5p expression showed an inverse relationship with centromere enumeration probe 17 (CEP17) signal count and correlated with membranous HER2 protein expression. hsa-miR-27b-5p expression was linked to CEP17 CNI, whereas hsa-miR-552-3p expression was associated with both increased HER2 amplification and CEP17 signal count. Importantly, hsa-miR-27b-5p upregulation independently predicted worse overall survival, whereas hsa-miR-128-3p upregulation independently predicted improved survival outcomes. These findings identify distinct microRNA signatures associated with HER2 pathway alterations and prognosis in gastric cancer, highlighting their potential as biomarkers and contributors to HER2-driven tumor biology. 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

Background: Age-related hearing loss (HL) is a significant independent risk factor for Alzheimer’s disease (AD), yet the molecular mechanisms underlying this comorbidity and the comparative efficacy of hearing interventions for cognitive outcomes remain unclear. This study aims to integrate clinical evidence and molecular data to address these gaps. Objective: The objective of this study was to conduct a systematic review and network meta-analysis (NMA) in order to: (1) compare the effects of hearing interventions on cognitive function in AD patients; (2) identify and rank key microRNAs (miRNAs) associated with AD-HL comorbidity; (3) explore heterogeneity sources; and (4) cross-validate findings with internal clinical sequencing data. Methods: We systematically searched PubMed, Web of Science, Embase, and the Cochrane Library, with a cut-off date of May 2024. Included studies involved AD patients with/without HL, reporting cognitive scores (MoCA, MMSE, and AVLT) or miRNA expression data. An NMA was performed to rank interventions (cochlear implants—CIs, hearing aids—HAs, and no intervention—NI) and miRNAs using surface under the cumulative ranking (SUCRA) curves. Heterogeneity was assessed via subgroup analysis and meta-regression. Pooled miRNA expression results were cross-validated against an internal clinical sequencing dataset (LC-P20240110033, n = 16) using the intraclass correlation coefficient (ICC) and Bland–Altman plots. Results: Twelve studies (2137 patients) were included. HL was significantly associated with worse cognitive function (MoCA: SMD = −0.82, 95% CI: −1.15 to −0.49; AVLT delayed recall: SMD = −1.12, 95% CI: −1.56 to −0.68). NMA revealed that the CI group (SUCRA = 0.89) was superior to the HA group (SUCRA = 0.62) and NI (SUCRA = 0.09) for preserving MoCA scores. Among the nine differentially expressed miRNAs identified in exploratory synthesis, three met strict quantitative criteria for NMA (reported in ≥2 independent studies with comparable quantification and variance data); hsa-miR-6875-5p was the most consistent biomarker (pooled FC = 1.52, 95% CI: 1.04–2.23), showing excellent agreement with sequencing data (FC = 3.29; ICC = 0.82, 95% CI: 0.67–0.91). Heterogeneity was significantly influenced by the miRNA detection platform (p = 0.04) and HL severity (p = 0.03). Conclusions: This study demonstrates that HL exacerbates cognitive decline in AD in a dose-dependent manner. Cochlear implants may offer superior cognitive protection compared to hearing aids. The consistently dysregulated hsa-miR-6875-5p emerges as a hypothesis-generating cross-modal biomarker, bridging clinical observation and molecular pathology in AD-HL comorbidity. Full article

Disease-modifying therapies (DMT)s) for relapsing-remitting multiple sclerosis (RRMS) act through distinct immunological mechanisms, yet the within-patient molecular response programs associated with these therapies remain incompletely defined. Here, we reanalyzed publicly available peripheral blood mononuclear cell (PBMC) miRNA microarray data (GSE230064) using a longitudinal, robustness-focused framework to compare therapy-associated miRNA response patterns following cladribine versus ocrelizumab treatment. Baseline (t0) and 6-month post-treatment (t1) samples were paired within individuals and technical replicates consolidated prior to analysis, yielding a final paired cohort of four cladribine-treated and six ocrelizumab-treated patients. Within each treatment arm, we quantified per-patient Δ-miRNA (t1 − t0) values and prioritized therapy-associated response features using a multi-evidence framework integrating effect direction, magnitude, directional consistency across individuals, and leave-one-out sensitivity. Cladribine treatment was associated with a highly coordinated, directionally concordant upregulation of five miRNAs including hsa-miR-27a-3p, hsa-miR-27b-3p, hsa-miR-503-5p, hsa-miR-148a-3p, and hsa-miR-26a-5p, all exhibiting 100% directional stability across patients and mean Δ-expression values ranging from +0.77 to +1.38. These miRNAs target pathways relevant to MS pathophysiology, including Th17/Treg balance, Wnt-β-catenin signaling, macrophage polarization, and epigenetic immune regulation. In contrast, ocrelizumab elicited a more selective response pattern, with five miRNAs including hsa-miR-100-5p, hsa-miR-410-3p, hsa-miR-432-5p, hsa-miR-296-5p, and hsa-miR-485-3p showing moderate directional stability (83%) and greater inter-individual heterogeneity, consistent with the more targeted mechanism of CD20+ B-cell depletion. Notably, the two treatment-associated signatures were non-overlapping, with hsa-miR-27b-3p representing the only miRNA shared with prior cross-sectional analyses of this dataset. The identified ocrelizumab-associated miRNAs implicate pathways including mTOR/IGF1R signaling, NF-κB regulation, RNA editing, and mitochondrial biogenesis, several of which are dysregulated in progressive MS. Together, these findings demonstrate that cladribine and ocrelizumab induce distinct, treatment-specific miRNA response architectures that reflect their divergent immunological mechanisms. This work establishes a stability-aware analytic template for extracting reproducible longitudinal miRNA signals from small paired RRMS cohorts and provides a ranked set of biologically plausible candidate miRNAs for prospective validation and mechanistic investigation. Full article

Although schistosome eggs are widely recognized as the principal drivers of hepatic granulomatous inflammation and fibrosis, the independent effects of adult worms may be masked by strong egg antigen-mediated responses. This study aimed to investigate whether adult Schistosoma japonicum worms alter the miRNA expression profile of hepatic stellate cells and to explore the potential relevance of these changes to liver fibrosis and hepatocellular carcinoma-related processes. A non-contact Transwell co-culture system was established using paired Schistosoma japonicum worms or male worms and hepatic stellate cells. Male worms were additionally included to further assess worm-derived effects independent of egg production–related influences. Untreated hepatic stellate cells served as controls. Total RNA was extracted for miRNA sequencing, and differentially expressed miRNAs were identified. Target gene prediction, Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis, and validation using The Cancer Genome Atlas database were subsequently performed. Both paired worms and male worms significantly altered the miRNA expression profile of hepatic stellate cells. Several differentially expressed miRNAs were identified, among which hsa-miR-103a-3p showed relatively stable changes. Pathway enrichment analysis suggested that the potential target genes of hsa-miR-103a-3p were mainly enriched in AMP-activated protein kinase, mechanistic target of rapamycin, tumor necrosis factor, insulin signaling, and cellular senescence pathways. Further analysis using The Cancer Genome Atlas database showed that hsa-miR-103a-3p had diagnostic value in hepatocellular carcinoma and was associated with alpha-fetoprotein level, albumin level, Ishak fibrosis score, pathological stage, histological type, and tumor status. These findings suggest that adult S. japonicum worms may alter the miRNA expression profile of hepatic stellate cells, and that hsa-miR-103a-3p may be associated with fibrogenic responses and may have potential relevance to hepatocellular carcinoma-related processes. However, this inference is based on correlative TCGA data and does not imply a causal role in schistosomiasis-associated hepatocarcinogenesis. Full article

Background and Objectives: MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression and have emerged as potential biomarkers in type 2 diabetes mellitus and its complications. This pilot exploratory study aimed to identify circulating miRNAs with differential expression in plasma from patients with newly diagnosed type 2 diabetes mellitus compared to age- and sex-matched healthy controls. Materials and Methods: Peripheral venous blood samples were collected from diabetic patients (n = 24) and controls (n = 12). Due to the exploratory nature of the study and limited sample material, samples were pooled within each group prior to plasma separation. Total RNA, including miRNAs, was extracted from plasma and analyzed using a high-throughput qPCR panel. Two normalization methods were applied to assess miRNA expression, and overlapping results were used for downstream analysis. Fold regulation was calculated using the 2^(−ΔCt) method. Results: A total of 33 and 42 miRNAs were identified as differentially expressed using the first and second normalization methods, respectively. Fourteen miRNAs were consistently downregulated across both methods. Several of these miRNAs, including hsa-miR-26a-5p, hsa-miR-146a-5p, hsa-miR-186-5p, hsa-miR-19a-3p, and hsa-miR-652-3p, have been previously associated with glucose metabolism, inflammation, and diabetic complications, such as retinopathy, neuropathy, and endothelial dysfunction. The pooling strategy enabled an efficient exploratory assessment of miRNA expression patterns while reducing inter-individual variability. Conclusions: This exploratory pilot study identifies a panel of circulating miRNAs with altered expression in pooled plasma samples from patients with newly diagnosed type 2 diabetes mellitus. These findings provide preliminary insights that warrant further validation in larger, individual-level studies to assess their diagnostic and prognostic potential. Full article

Non-small cell lung cancer (NSCLC) remains a major cause of cancer-related mortality, with poor survival outcomes despite advances in surgery, chemotherapy, targeted therapy, and immunotherapy. The tumor microenvironment (TME) plays a central role in sustaining tumor growth, immune evasion, and systemic metabolic dysfunction. In this study, we performed an integrative analysis of differentially expressed microRNAs (miRNAs) to uncover their contributions to dysregulated signaling networks in NSCLC. hsa-miR-486-5p was identified as a prominent differentially expressed candidate miRNA. Using mathematical modeling and regression-based reduction, we identified Forkhead Box O1 (FOXO1) and Unc-51 like Autophagy Activating Kinase 2 (ULK2) as critical regulatory nodes that integrate oncogenic signaling with cellular homeostasis. Aberrant expression of hsa-miR-486-5p was found to modulate pathways including PI3K/AKT/mTOR, NF-κB, and JAK-STAT3, thereby promoting tumor progression and secretion of inflammatory cytokines. These cytokines, viz., IL-6, TNF-α, and IL-1β, activate muscle-specific protein degradation pathways through E3 ubiquitin ligases TRIM63 and FBXO32, linking NSCLC progression to cancer-associated sarcopenia. Quasipotential landscape analysis further revealed dynamic phenotypic transitions between stable and unstable states, highlighting the adaptability of tumor–host interactions. Collectively, our findings demonstrate that miRNA-mediated regulatory networks not only drive NSCLC progression and inflammation but also contribute to systemic muscle wasting. These insights emphasize the need for novel therapeutic strategies, including RNA-based interventions, to overcome resistance, improve survival, and address the metabolic complications associated with NSCLC. Full article

Background: Postmenopausal osteoporosis is associated with cellular senescence and the accumulation of the senescence-associated secretory phenotype (SASP). While mesenchymal stem cell (MSC)-derived exosomes show tissue repair potential, the efficacy and mechanisms of MSC-derived apoptotic vesicles (apoVs) remain unclear. This study compared MSC-apoVs and exosomes in postmenopausal osteoporosis and investigated the underlying epigenetic mechanisms. Methods: Therapeutic efficacy was evaluated in an ovariectomized (OVX) mouse model and senescent human bone marrow mesenchymal stem cells (hBMMSCs). Small RNA sequencing identified differential microRNA (miRNA) cargos between vesicle types. SASP-related cytokine expression (IL-6, TNF-α, MCP-1) and pathway activation were assessed by RT-qPCR, ELISA, and Western blot. Results: MSC-apoV treatment attenuated bone loss in OVX mice and reduced SASP expression in senescent hBMMSCs to a greater extent than exosomes. Small RNA sequencing revealed that apoVs were enriched with a specific miRNA cluster, including hsa-let-7b-5p, hsa-miR-92a-3p, and hsa-miR-98-5p. Bioinformatic analyses identified BRAF and CRKL as downstream targets of this miRNA cluster, supported by reduced protein levels after apoV treatment. Subsequent molecular assays showed that apoV treatment inhibited the phosphorylation of both the MAPK (p38 and JNK) and NF-κB (p65) signaling pathways, which correlated with reduced local inflammation in the bone marrow microenvironment and preserved osteogenic differentiation capacity. Conclusions: MSC-apoVs attenuate postmenopausal osteoporosis more effectively than exosomes. This enhanced efficacy is associated with the delivery of an enriched miRNA cluster that inhibits MAPK and NF-κB signaling, together with suppression of BRAF and CRKL protein expression. ApoVs may represent a cell-free therapeutic strategy for age-related bone loss. Full article