Search Results (454)

Background/Objectives: Type 1 diabetes (T1D) is an autoimmune disease characterized by progressive β-cell loss. Apolipoprotein CIII (apoCIII), a lipid metabolism regulator, is elevated in T1D and implicated in β-cell apoptosis. Antisense oligonucleotide–mediated apoCIII reduction delays diabetes onset in diabetes-prone BioBreeding (DPBB) rats. This study examined whether small-interfering RNA (siRNA) targeting apoCIII during the final prediabetic month preserves islet integrity and delays T1D onset. Methods: Two siRNAs targeting rat apoCIII were evaluated in 30-day-old DPBB rats for efficacy and off-target effects. Hepatic and plasma apoCIII levels were measured, and neighboring apolipoprotein gene expression was assessed. The most specific candidate (apoCIII-siRNA2) was selected. Duration of action was determined after a single injection. To study the effects of apoCIII-lowering treatment in vivo, islets from 25-day-old DPBB rats were transplanted into the anterior chamber of the eye of age-matched DPBB recipients. Rats received weekly intravenous injections of apoCIII-siRNA2 from day 30 until diabetes onset. Islet morphology, vascularization, and phagocyte infiltration were assessed by confocal imaging three and five weeks post-transplantation. Results: Both siRNAs reduced apoCIII, but one showed off-target effects and was excluded. A single injection of apoCIII-siRNA2 suppressed plasma apoCIII for approximately one week and weekly treatment maintained low circulating apoCIII levels. Five weeks after transplantation islet morphology and vascularization were preserved, and there was no increase in phagocyte infiltration. This resulted in a delayed onset of diabetes. Conclusions: siRNA-mediated apoCIII reduction delays pancreatic islet deterioration and T1D onset in DPBB rats, supporting apoCIII as a contributing factor to β-cell vulnerability and thereby a potential therapeutic target. Full article

Background: MicroRNAs (miRNAs) are small, non-coding RNA molecules that regulate gene expression post-transcriptionally by binding to target messenger RNAs (mRNAs) and suppressing their expression. Competing endogenous RNAs (ceRNAs), including mRNAs and circular RNAs (circRNAs), modulate miRNA availability through competitive binding, forming regulatory networks that fine-tune gene expression. CircRNAs can act as miRNA sponges, reducing miRNA-mediated repression of other targets, a mechanism implicated in various pathophysiological processes, including oncogenesis. Methods: We propose a mathematical model describing the dynamics of miRNA–mRNA–protein interactions, extending existing frameworks for miRNA–mRNA regulation. A qualitative analysis of the associated nonlinear differential equations system is performed. Results: We prove the boundedness of all positive solutions, establish the existence of a unique positive attracting equilibrium, and provide a mathematical perspective on the crosstalk mechanism in protein production. Conclusions: The effectiveness of ceRNA interactions depends on the relative abundance of miRNAs and their targets. This highlights the ongoing debate regarding the biological impact of low-abundance RNA transcripts on miRNA-mediated regulation. Full article

Arrhythmogenic cardiomyopathy (ACM/ARVC) is an inherited myocardial disease characterized by progressive fibro-fatty replacement, ventricular arrhythmias, and an increased risk of sudden cardiac death. In addition to mutations in desmosomal genes, growing evidence suggests that microRNAs (miRNAs) actively contribute to disease pathogenesis by regulating key processes such as fibrosis, cell adhesion, and cardiac remodeling. This systematic review analyzed the main miRNAs identified in studies of human cardiac tissue and animal models of ARVC. Materials and Methods: Studies based on human myocardial tissue analysis (including autopsy and biopsy samples) and animal models of arrhythmogenic cardiomyopathy were included, using RNA sequencing, small RNA sequencing, miRNA arrays, and RT-qPCR. Studies on circulating miRNAs and narrative reviews were excluded. miRNAs were analyzed in relation to their functional pathways and their role in disease pathogenesis. Results: The synthesis of studies on human and animal cardiac tissue reveals a consistent miRNA signature associated with arrhythmogenic cardiomyopathy. MiR-21-5p and miR-29b-3p are associated with fibrosis and extracellular matrix remodeling, whereas miR-133a-b and miR-130a are linked to cardiomyocyte integrity loss and desmosomal dysfunction. A second group of miRNAs, including miR-217-5p, miR-708-5p, and miR-135b, regulates key pathways such as Wnt/β-catenin and Hippo signaling, contributing to structural remodeling and loss of cellular identity. Furthermore, downregulation of miR-499-5p is associated with mitochondrial dysfunction and cellular vulnerability, while the miR-142-3p, miR-182-5p, and miR-183-5p clusters contribute to differential molecular signatures compared with other cardiomyopathies. Overall, miRNAs converge on three main pathogenic axes: myocardial fibrosis, desmosomal impairment, and remodeling of cellular signaling pathways. Conclusions: The available evidence indicates that arrhythmogenic cardiomyopathy is regulated by a coordinated network of miRNAs that actively drives myocardial damage progression. These miRNAs represent not only biomarkers but also functional mediators of disease, suggesting potential diagnostic and therapeutic applications based on tissue-specific molecular signatures, including in post-mortem settings. Full article

Small extracellular vesicles (sEVs) are extracellular vesicles that mediate intercellular communication through the transfer of bioactive molecules, including proteins, lipids, and nucleic acids. Among their cargo, microRNAs (miRNAs) have emerged as critical regulators of gene expression with significant implications in cancer biology. Tumor-derived extracellular vesicle-associated microRNAs (EV-miRNAs) can reprogram recipient cells, promoting oncogenesis, metastasis, angiogenesis, and therapeutic resistance. This review provides a comprehensive overview of EV-miRNAs in cancer, examining their biogenesis, mechanisms of intercellular transfer, and functional roles in tumor progression. We discuss the clinical potential of EV-miRNAs as non-invasive biomarkers for cancer diagnosis and prognosis, as well as their emerging applications in targeted therapeutic strategies. Furthermore, we address current challenges related to isolation techniques, quantification methods, and standardization protocols that hinder clinical translation. Finally, we outline future directions for integrating EV-miRNA analysis into precision oncology frameworks and liquid biopsy platforms, highlighting opportunities to advance personalized cancer care. Full article

Mitochondrial dysfunction is not merely a byproduct of transformation but a driver of tumorigenesis, metastasis, and therapeutic resistance. Recent advancements in intercellular communication have identified Extracellular Vesicles (EVs) or exosomes as critical mediators that bridge the gap between the tumor and its microenvironment (TME). These EVs contain a complex repertoire of bioactive cargo, including proteins, lipids, and RNAs. Among the class of RNAs, small non-coding RNAs, microRNAs (miRNAs), are the most abundantly expressed bioactive compounds that are selectively packaged and delivered to recipient cells. EV-delivered miRNAs can target nuclear-encoded mitochondrial genes and have also been reported to localize to mitochondria (mitomiRs), where they function as post-transcriptional regulators of bioenergetic and mitochondrial dynamic adaptations that support tumor progression. This review explores the “EV-miRNA-Mitochondria Axis”, delineating the molecular mechanisms by which EV-carried miRNAs reprogram the “Mitochondrial Information Processing System” (MIPS) - a signaling network where mitochondria integrate metabolic cues (e.g., ROS, calcium flux) to dictate critical biological outcomes, such as immune regulation and cell survival. We summarized specific sorting machineries (e.g., hnRNPA2B1, Lupus La) that package oncogenic miRNAs into EVs and how these cargoes hijack mitochondrial function upon delivery. Specifically, we discussed how EV-miRNAs induce metabolic shifts, manipulate mitochondrial dynamics (fission/fusion), and inhibit the intrinsic apoptosis to drive cancer progression. Finally, we highlighted the dual utility of these EV-miRNAs as drivers of pathogenesis and promising non-invasive biomarkers for early diagnosis, prognostic and therapeutic monitoring. Full article

Transposable elements (TEs) are increasingly recognized as modulators of innate immunity, yet their antiviral functions remain poorly understood outside mammals and dipterans. Here, we identify a long terminal repeat retrotransposon, LmGypsy, as a key regulator of antiviral defense in Locusta migratoria. The infection of Acrididae reovirus (ARV) induces rapid upregulation of LmGypsy, and its inhibition compromises antiviral resistance. Mechanistically, LmGypsy promotes viral-derived DNA (vDNA) production, which drives Dicer-2-dependent biogenesis of virus-derived small interfering RNAs (vsiRNAs) to enhance RNA interference-mediated viral clearance. Notably, vDNA persists throughout infection, suggesting a role in sustaining antiviral responses. In parallel, LmGypsy activity is positively associated with induction of cyclic GMP-AMP synthase (cGAS)-like receptors (LmcGAS1/2/4) and their downstream effector Stimulator of Interferon Genes (STING). Together, these findings support a dual-layer antiviral strategy and indicate that TE-mediated immunity represents a widespread antiviral mechanism across taxa. Full article

MicroRNAs (miRNAs) are small non-coding RNAs that negatively regulate gene expression at the post-transcriptional level. In plants, miRNAs are involved in environmental responses and can be transferred to other species to mediate cross-kingdom regulation of gene expression. This mechanism has recently been proposed in mammals, yet evidence remains scarce and inconsistent. Multiple studies have shown that a fraction of plant-derived miRNAs (pmiRNAs) present in food resist processing, cooking, and digestion. Evidence suggests that dietary-derived pmiRNAs might be absorbed in the gastrointestinal tract and enter circulation, predominantly packaged in extracellular vesicles, and reach different tissues, where they might exert cross-kingdom gene expression regulation. Nonetheless, several attempts to reproduce or confirm the above results have failed, raising questions about the reliability of studies supporting the hypothesis of cross-kingdom gene regulation by pmiRNAs. In this review, we recapitulate the state-of-the-art knowledge in the field, addressing both supporting and opposing evidence, as well as the main analytical challenges that need to be taken into consideration, in an effort to provide a comprehensive framework on the controversial evidence collected so far and support the use of best practices for future research. Full article

Background/Objectives: Osteosarcoma is the most common primary malignant bone tumour, affecting children and young adults. Recent evidence suggests that extracellular vesicles (EVs), small membrane-bound nanoparticles released by all cell types, play a key role in intercellular communication within the tumour microenvironment. Therefore, we aimed to investigate the effects of osteoblast-derived EVs (OB-EVs) on osteosarcoma cell behaviour and to characterise the transcriptional and miRNA-mediated mechanisms underlying these effects. Methods: Phenotypic assays were performed to assess metabolic activity, proliferation, apoptosis, and invasion ability of human osteosarcoma cell lines after treatment with OB-EVs. Illumina-based RNAseq was conducted on RNA isolated from OB-EVs-treated cells, and qRT-PCR was assessed using commercially available TaqMan miRNA cards on RNA isolated from OB-EVs. Results: In U2OS cells, OB-EVs reduced metabolic activity (1.30-fold decrease, p = 0.0137) and proliferation (1.70-fold decrease, p = 0.017) while increasing apoptosis (1.15-fold increase, p = 0.014). In MG63, OB-EVs increased proliferation (4.9-fold increase, p = 0.020) without affecting tumour cell aggressiveness, while normal osteoblast behaviour was not affected by OB-EVs. MNNG/HOS cells treated with OB-EVs for 48 h showed substantial transcriptomic changes, with 296 differentially expressed genes (97 up- and 199 down-regulated in OB-EVs treated cells versus untreated cells), indicating a direct impact of OB-EVs on gene expression. Intriguingly, Gene Set Enrichment Analysis (GSEA) showed trends consistent with modulation of signalling pathways, including Wnt/β-catenin and NOTCH. Conversely, miRNA profiling of OB-EVs identified 13 highly expressed miRNA. Integration of transcriptomic and miRNA target prediction data highlighted convergent pathway-level signals, suggesting that OB-EVs may modulate tumour-associated regulatory networks. Conclusions: Taken together, these findings indicate that OB-EVs modulate osteosarcoma cell phenotype, with miRNA shuttling representing a potentially relevant contributing mechanism. The integrative analysis suggests that pathways associated with proliferation and cellular homeostasis, including Wnt/β-catenin signalling, may be involved, although further functional validation is required to confirm these mechanisms. Full article

Intensifying drought stress under global climate change poses a significant threat to woody plants, highlighting the critical need to identify key genes conferring drought tolerance. Here, we characterized PsnSAUR6, a Small Auxin Upregulated RNA (SAUR) family gene from poplar (Populus simonii × P. nigra) that is responsive to drought and abscisic acid (ABA). Overexpression of PsnSAUR6 in transgenic tobacco conferred superior drought tolerance, evidenced by increased biomass, enhanced root elongation, improved stomatal regulation, and favorable physiological responses, including higher proline content and peroxidase (POD) activity but lower malondialdehyde (MDA). Transcriptome analysis revealed that under water deficit, PsnSAUR6 suppressed the ABA negative regulator PP2C37 while upregulating key antioxidant defense-related transcription factors (ERF020, NAC83, MYB2) and the potassium transporter HAK5. Collectively, these findings establish PsnSAUR6 as a positive regulator in ABA-mediated drought adaptation, presenting it as a promising genetic target for enhancing the climate resilience of woody plants. Full article

Background/Objectives: Triple-negative breast cancer (TNBC) is frequently characterized by notably elevated Ki-67 expression, a hallmark of uncontrolled rapid cell-cycle progression. However, the underlying mechanisms remain unclear, leading to limited therapeutic options. Methods: In this study, hub gene was identified through integrated bioinformatic analysis of public datasets (TCGA-BRCA and METABRIC). Subsequent functional validation was performed both in vitro and in vivo using siRNA-mediated knockdown and small-molecule inhibitors. Phenotypic effects—including cell viability, cell cycle distribution, DNA synthesis, and clonogenic survival—were comprehensively assessed using MTT assays, flow cytometry, EdU, and colony formation assays. Protein-level changes were confirmed by Western blotting and immunohistochemistry (IHC). To dissect the transcriptional regulation of the key hub gene TTK, we first predicted potential upstream transcription factors using the JASPAR database; binding specificity was then validated through in silico motif analysis, luciferase reporter assays, and chromatin immunoprecipitation followed by quantitative PCR (ChIP-qPCR). Results: The mitotic kinase TTK is significantly overexpressed in TNBC compared with non-TNBC breast cancers. Notably, TTK overexpression exhibited a strong positive correlation with elevated Ki-67 indices and reduced overall survival in TNBC patients. Functional validation demonstrated that pharmacological or genetic inhibition of TTK effectively induced G2/M cell-cycle arrest and potently suppressed TNBC proliferation in both in vitro cell cultures and in vivo xenograft models. Mechanistically, TTK overexpression stems from enhanced transcriptional initiation driven by the transcription factor NFYA binding to the CCAAT box in the TTK promoter—an interaction newly identified here. Concurrently, TTK blockade disrupted spindle assembly checkpoint (SAC) signaling via BUB1B/MAD1L1 downregulation, triggering mitotic arrest and catastrophe. Conclusions: Collectively, these findings establish TTK as a key cell-cycle regulator driving TNBC proliferation. More importantly, targeting mitotic control through TTK inhibition represents an efficient strategy to impede the aberrantly fast cell cycle progression in TNBC. Full article

Extracellular vesicles (EVs) from the edible mushroom Pleurotus tuber-regium (PTR) were investigated with respect to their environmental responsiveness, molecular features, and preliminary functional properties. PTR-EVs were characterized by dynamic light scattering, nanoparticle tracking analysis, and transmission electron microscopy. Proteomic analysis revealed enrichment of ribosomal and proteasomal proteins, redox-related enzymes, and vesicle trafficking components, suggesting non-random molecular representation. Small RNA sequencing identified abundant novel miRNAs with predicted targets involved in nitrogen metabolism, cell wall remodeling, redox regulation, and ubiquitin-mediated proteolysis. Among the tested factors, temperature showed the strongest association with vesicle production, with particle concentration increasing from 1.22 × 10⁹ to 7.31 × 10⁹ particles/mL at 34 °C, approximately six-fold higher than at 30 °C. Transcriptomic profiling showed coordinated repression of cell wall-associated genes and redox enzymes, together with induction of endoplasmic reticulum proteostasis pathways, consistent with stress-associated changes in the cellular context of vesicle release. Ultrasonicated PTR-EVs exhibited enhanced DPPH and ABTS radical-scavenging activities in chemical assays, with DPPH increasing from 59.52% to 71.73% and ABTS from 38.25% to 40.51%. Encapsulation efficiencies reached 32.67% ± 1.3% for proanthocyanidins and 46.01% ± 0.5% for curcumin. PTR-EVs showed the best short-term stability at pH 7 and 4 °C, supporting their further evaluation as an edible fungal vesicle platform for food-related nanoscale delivery. Full article

Sweet–waxy maize is a highly valuable specialty maize type with an increasing market demand, but conventional breeding methods for producing sweet–waxy maize are restricted by severe bottlenecks, such as long breeding cycles and linkage drag. This study was conducted to rapidly create sweet–waxy maize germplasm using CRISPR/Cas9 genome-editing technology. We used a CRISPR/Cas9 system to target maize Sh2 (regulating the super-sweet kernel trait) and Wx (controlling the waxy kernel trait), which are two key genes in the starch biosynthesis pathway. Two small-guide RNAs (sgRNAs) designed for each gene were incorporated into CRISPR/Cas9 vectors, which were then introduced into maize via Agrobacterium-mediated transformation. We obtained Cas9-free T₃ homozygous sh2 and wx mutant lines with significant increases in kernel soluble sugar and amylopectin contents, respectively, but no adverse changes to major agronomic traits. Using these Cas9-free lines, we developed a new type of sweet–waxy maize germplasm, in which waxy and sweet kernels on the same ear segregated at a 3:1 ratio. Our results indicate that CRISPR/Cas9-mediated editing of Sh2 and Wx can efficiently generate sweet–waxy maize germplasm with no detectable linkage drag. The study methods would be useful for optimizing the molecular breeding of novel and innovative maize germplasm. Full article

Non-coding RNAs (ncRNAs) have emerged as important regulators of gene expression and cellular homeostasis, and their dysregulation is now recognized as a hallmark of cancer. Over the past decades, extensive research has demonstrated that diverse ncRNA classes, including microRNAs (miRNAs), long non-coding RNAs (lncRNAs), circular RNAs (circRNAs), and other small ncRNA species, participate in complex regulatory networks that influence tumor initiation, progression, metastasis, and therapy response. Through mechanisms such as transcriptional regulation, post-transcriptional gene silencing, epigenetic modulation, and competitive endogenous RNA interactions, ncRNAs shape the molecular circuitry underlying cancer development. In addition to their functional roles in tumor biology, many ncRNAs are released into biological fluids and can be detected as circulating molecules in blood, urine, saliva, and other biofluids. Their remarkable stability in extracellular environments has generated considerable interest in their use as minimally invasive biomarkers in liquid biopsy applications. Emerging evidence has shown that circulating ncRNAs (c-ncRNAs) can support cancer detection, disease stratification, and treatment monitoring. This narrative review provides an integrated view that links ncRNA-mediated regulatory networks with their application as liquid biopsy biomarkers, positioning ncRNAs as comprehensive indicators of tumor conditions. Particular emphasis is placed on c-ncRNA biomarkers, the integration of multiple ncRNA classes, and multi-analyte biomarker strategies that combine ncRNAs with complementary circulating molecules such as cell-free DNA and protein markers. Finally, we discuss the technical and clinical challenges that currently limit the translation of ncRNA-based diagnostics into clinical practice and highlight future directions for advancing ncRNA-guided liquid biopsy approaches in precision oncology. Full article

Triple-negative breast cancer (TNBC) is an aggressive subtype lacking targeted therapies and characterized by pronounced heterogeneity and widespread dysregulation of microRNAs (miRNAs) that influence epithelial-to-mesenchymal transition (EMT) and metastasis. Tumor-derived extracellular vesicles (tEVs) further contribute to TNBC progression by transporting oncogenic cargo that can enhance pro-inflammatory signaling. The synthetic SMRwt peptide has been suggested to modulate oncogenic pathways; however, its effects on EV miRNA composition and inflammatory transcript profiles in TNBC remain unclear. Here, we investigated whether SMRwt alters tEV-associated miRNAs and cytokine transcript signatures relevant to EMT and inflammasome-linked pathways. Extracellular vesicles were isolated from SMR-treated and untreated MDA-MB-231 cells, followed by nanoparticle tracking analysis and small RNA sequencing. SMRwt treatment enriched 11 tumor-suppressive miRNAs (including Let-7a-5p, Let-7b-5p, miR-24-3p, miR-26b-5p, miR-92a-3p, miR-93-5p, and miR-496) previously associated with the regulation of proliferation, EMT, migration, and metastasis. We also observed modest, non-significant decreases (1.01–1.27-fold) in oncogenic miR-1200, miR-374a-5p, and miR-937-3p, which have been implicated in the progression of breast, lung, and bone malignancies. Complementary transcriptomic profiling using the NanoString nCounter Breast Cancer 360 Gene Expression Panel (NanoString Technologies, Inc., Seattle, CA, USA) demonstrated reduced expression of inflammasome-associated cytokines in TNBC cells relative to non-tumorigenic controls, including a log₂ fold change of −1.15 for IL 1β (MDA-MB-231 vs. MCF10A). These transcript-level changes suggest potential modulation. Additionally, SMRwt suppresses ASC-mediated caspase-1 activation and reduces IL-1β secretion, thereby inhibiting NLRP3 inflammasome signaling. Therefore, we infer that SMRwt simultaneously restores tumor-suppressive miRNA networks and suppresses inflammasome-driven inflammation, supporting its potential as a dual-target therapeutic strategy for TNBC. Full article

MicroRNAs (miRNAs) are small non-coding RNAs that play central roles in post-transcriptional gene regulation and cellular homeostasis maintenance. Dysregulation of miRNA expression is increasingly recognized as a key contributor to tissue injury during the acute phase and to disease progression in the chronic phase. Chronic kidney disease (CKD) commonly progresses and ultimately leads to kidney failure through interstitial fibrosis, which is the final common pathway of CKD progression. Interstitial fibrosis is driven not only by fibroblast activation but also by phenotypic transitions in injured tubular epithelial cells, infiltrating macrophages, and peritubular capillary cells. These multifaceted cellular pathways induce and exacerbate interstitial fibrosis, and several miRNAs have been identified as important regulators of these pathways. In addition to fibrotic pathophysiological features, disease-specific dysregulation of miRNAs has been increasingly detected in various causes of CKD, including diabetic kidney disease, chronic glomerulonephritis, and nephrosclerosis. In this review, we provide an integrated overview of miRNA-mediated regulation in CKD, with particular emphasis on cell lineage functions within fibrotic pathways and disease-specific roles. Finally, we discuss the emerging potential of miRNAs as biomarkers and therapeutic targets for CKD and highlight future research directions. Full article