m6A is the predominant internal RNA modification in eukaryotic cells and is distinguished by its abundance and evolutionary conservation. This epigenetic mechanism is dynamically controlled by a coordinated system of writer, eraser, and reader proteins. This sophisticated posttranscriptional regulatory mechanism precisely controls gene expression by influencing RNA metabolism, including its stability, translation, and splicing. Recent advances have revealed the functions of m6A in female reproductive cancers, early embryonic development, and stem cell differentiation. However, its functional roles and molecular mechanisms throughout pregnancy and in related disorders remain incompletely understood, which, to some extent, limits its clinical translation. This review systematically outlines the core regulators of m6A, advanced detection technologies, and its regulatory network across the continuum of pregnancy. Given the immunological parallels between the maternal-foetal interface and the tumour microenvironment, we discuss the possible function of m6A modifications in regulating the maternal-foetal immune microenvironment. The aims of this review were to elucidate the m6A regulatory network across gestation and evaluate its potential as a source of diagnostic biomarkers and therapeutic targets for pregnancy-related pathologies.

Cancer stem cells (CSC) can sustain tumor growth and therapeutic resistance in part through their contribution to vasculogenic mimicry (VM) in ovarian cancers. Pharmacological targeting of CSC-associated transcriptional programs could represent a promising strategy to overcome recurrence and metastasis. While preclinical studies show caffeic acid phenethyl ester (CAPE), a plant-derived metabolite, can sensitize tumors to chemotherapy and radiotherapy, little is known about its anti-VM properties.

Chronic cigarette smoke (CS) disrupts epithelial homeostasis, fuels persistent inflammation, and impairs alveolar repair-hallmarks of COPD with few disease-modifying options. Extracellular vesicles (EVs) from human umbilical cord mesenchymal stem cells (hUC-MSCs) are emerging as cell-free modulators of regeneration, yet their impact on the CS-injured alveolus and alveolar type-2 (AT2) stem/progenitor programs remains unclear. We used a preclinical model of chronic CS exposure coupled with organoid-guided analyses to test whether hUC-MSC-derived EVs can restore epithelial regeneration while tempering injury-associated inflammation and remodeling. Following CS injury, animals received vehicle, hUC-MSCs, or purified hUC-MSC EVs; lungs were evaluated histologically (airway/parenchymal inflammation, emphysema-like change), by Masson's trichrome (collagen deposition), and functionally using ex vivo epithelial organoids (organoid number/size, architecture, and AT2/AT1 marker balance). Transcriptomic profiling of organoid-derived RNA mapped pathway-level changes. CS induced robust immune-cell infiltration, increased collagen, and abnormal organoid phenotypes consistent with dysregulated progenitor activity. Post-injury EV treatment reduced inflammatory infiltrates and collagen, normalized organoid number and size, and restored AT2/AT1 lineage balance toward naïve patterns. At the molecular level, EVs dampened injury-upregulated circuits (including IL-17, PI3K-AKT-mTOR, MAPK, oxidative-stress and matrix-remodeling signatures) and enriched pathways associated with epithelial homeostasis and barrier integrity. Together, these data position hUC-MSC EVs as precision modulators of the injured alveolar niche that rebalance inflammation and re-engage endogenous regenerative programs. The organoid-guided, multi-scale readouts provide mechanistic insight and a translational rationale for EV-based regenerative therapeutics in smoke-induced lung injury and, by extension, COPD.

Hard-to-soft tissue interfaces, such as bone-tendon or bone-ligament junctions, remain a challenge to treat. Low healing success rates stem from the complexities at the interface, creating an urgent need for better models to elucidate the properties that enable these junctions to withstand complex mechanical loads and to function as hubs for crosstalk among different cell populations.

Oncolytic viruses (OVs) are widely studied for their ability to lyse cancer cells and prime immune responses; however, the immune consequences triggered by OVs remain incompletely understood. Here, we discover that oncolytic VSVΔ51 treatment suppresses the T cell receptor signaling of tumor-infiltrating T cells. Mechanistically, VSVΔ51-infected cancer cells upregulate PCSK9 secretion, which triggers lysosomal degradation of major histocompatibility complex (MHC)-I in bystander cells. PCSK9 inhibition synergizes with VSVΔ51 treatment to suppress tumor growth in multiple colorectal cancer models and induce complete regression in a microsatellite-stable (MSS) tumor model. This combination fosters stem-like CD8+ T cells and establishes anti-tumor memory. Engineered VSVΔ51 expressing anti-PCSK9 single-chain variable fragments improves intra-tumor viral replication, sustains anti-tumor CD8+ T cell memory, and enhances anti-PD-1 therapy efficacy. Our results identify the role of PCSK9 in the immunosuppressive feedback following viral infection and propose a strategy for engineered oncolytic virotherapy.

Orofacial and limb muscles differ in embryonic origin and regenerative capacity. Neuromuscular junction (NMJ) regeneration is critical for muscle restoration both histologically and functionally. The relative potential of orofacial and limb muscles to form postsynaptic apparatuses remains elusive. While the role of fibro-adipogenic progenitors (FAPs) in NMJ regeneration has been discussed in limb muscles, it remains unexplored in orofacial muscles.

Fragile X syndrome (FXS) is a neurodevelopmental disorder caused by loss of fragile X messenger ribonucleoprotein (FMRP). In most cases, this results from a CGG expansion exceeding 200 repeats in the 5' untranslated region of the fragile X messenger ribonucleoprotein 1 (FMR1) gene, known as the "full mutation". While the trinucleotide expansion has long been thought to induce epigenetic silencing of this locus, studies have shown that many males with a full mutation still express FMR1 mRNA. However, these individuals produce little to no FMRP protein, due to mechanisms that remain unclear. Mis-splicing of FMR1 transcripts with an expanded CGG tract has recently been proposed as a potential mechanism underlying the absence of FMRP in FXS tissues despite the presence of gene transcripts.

The Jiangquan black pigs, a new breed of swine obtained by introducing traits from Duroc pigs into Yimeng black pigs, exhibits fast growth rates and high meat quality. To understand how daily weight gain influences muscle development in this breed, we analyzed longissimus dorsi muscle cell subpopulations from Jiangquan black pigs using snRNA and bulk RNA sequencing. Thirteen distinct cell types (e.g., muscle stem cells, satellite cells, fibroblasts) were identified, and marker genes (PAX7, MYOD, MYOG) were found to exhibit stage-specific expression during differentiation. Pseudotime analysis revealed the differentiation trajectories of these cell populations, while cell cycle analysis uncovered the higher mitotic activity in satellite cells of the fast-growth versus slow-growth groups. Furthermore, cell communication analysis highlighted the interactions between muscle cells and other cell types. Finally, intergroup analysis revealed that 2,466 and 2,597 genes were differentially expressed in muscle stem cells and muscle satellite cells, respectively. These genes were enriched in disease-related pathways. This study provides a single-cell resolution atlas of porcine muscle development, offering insights into the genetic regulation of growth and potential targets for breeding optimization.

Mechanical stimulation plays a crucial role in odontoblast differentiation. However, the underlying molecular mechanisms remain unclear. We have previously shown that hydrostatic pressure (HP) applied to stem cells from human exfoliated deciduous teeth (SHED) promotes odontoblast differentiation by translocating RUNX2 and increasing WNT16 expression through PIEZO1 signaling. In this study, we further explored the downstream signaling cascade linking PIEZO1 activation and odontoblast differentiation. HP stimulation increased the expression of odontoblast differentiation markers PANX3 and DSPP, as shown by qPCR, and enhanced Alizarin Red staining-results significantly suppressed by siRNA targeting either PIEZO1 or WNT16. RT-PCR analysis revealed that, among the two known human WNT16 isoforms, only WNT16b was expressed in SHED. qPCR demonstrated that HP-induced WNT16 expression was reduced by siPIEZO1 and further decreased by siRUNX2. Promoter analysis identified four RUNX2-binding sites within the upstream region of WNT16. A luciferase reporter assay using plasmids containing the WNT16 promoter showed that RUNX2 overexpression in HEK293 cells significantly increased luciferase activity. Moreover, HaloChIP assays with a HaloTag-RUNX2 expression vector confirmed RUNX2's binding to the WNT16 promoter. These findings suggest that PIEZO1-mediated mechanical stress promotes odontoblast differentiation through the RUNX2-dependent transcriptional activation of WNT16.

Inflammatory bowel disease (IBD) is secondary to an abnormal immune response to the microbiota. To study this, models of host-microbe interactions that represent mucosal bacterial communities and inter-patient diversity are required. Human intestinal organoids (HIOs) are an established model to investigate epithelial responses. Here, we describe a technique of culturing bacteria directly from the sites of inflammation in IBD, while simultaneously sampling host tissue. We generated HIOs from a cohort of newly diagnosed paediatric IBD patients, without confounding treatments or comorbidities, and explored their response to site-specific bacteria. A unique biobank of matched HIOs and cultured mucosa-attached bacteria was established from 27 paediatric patients. Transcriptional profiling revealed differential gene expression between control and IBD-derived organoids. We used microinjection to introduce bacteria to the apical surface of the epithelium, to determine the effect of bacteria on host epithelial cells. We measured survival and growth of bacteria within the HIOs and tested several related bacterial isolates for their impact on the epithelium. An isolate from a control patient stimulated inflammatory signalling pathways but this was not observed in response to a closely related isolate originating from an IBD patient. This study demonstrates the feasibility of isolating bacteria and generating organoids from the same biopsy tissue, to explore personalised host-microbe interactions. The microinjections, while labour-intensive, demonstrate that closely related bacteria can induce very different epithelial responses, with downstream implications for immune response. This highlights the importance of understanding host-microbe interactions in a strain- and site-specific manner and developing techniques for personalised microbiome-based therapeutics.

Bladder cancer is a common urogenital malignancy that remains difficult to treat, particularly due to therapeutic resistance, such as resistance to cisplatin, in which cancer stem cells (CSCs) play a central role. This study investigates the combination of 5-aminolevulinic acid-mediated photodynamic therapy (5-ALA-PDT) and berbamine as a potential multimodal treatment strategy using the bladder cancer cell lines RT112 and J82, their cisplatin-resistant variants, and generated CSC-like cells. Berbamine is a natural plant compound and was confirmed in this study to have anticancer properties by inhibiting cell migration and invasion, and by inducing apoptosis. This study also showed that berbamine enhances the accumulation of protoporphyrin IX (PpIX), the photosensitizer induced by 5-ALA. 5-ALA-PDT destroys cancer cells by stimulating PpIX via 635 nm red laser light to produce reactive oxygen species (ROS). This was found to happen in all tested cell lines, whereas berbamine could modulate the cell destruction in a concentration-dependent manner and was influenced by the specific biological characteristics of the tested cell variants. CSCs showed the strongest response to the combination therapy approach, suggesting that they may represent more vulnerable cell variants to the tested treatment. Cisplatin-resistant cell lines could also be treated successfully with 5-ALA-PDT, whereas berbamine could enhance its efficacy in the cisplatin-resistant J82 LTT. These findings suggest that the combination treatment of 5-ALA-PDT and berbamine may serve as a promising approach to overcome therapeutic resistance in bladder cancer, particularly in cisplatin-resistant and CSC-enriched tumour types.

Gastrointestinal dysfunction often precedes motor symptoms in Parkinson's disease (PD), suggesting the enteric nervous system (ENS) is central to early pathogenesis. How α-synuclein contributes to ENS dysfunction, and how inflammation modulates this, remains unclear. Here we show that Tumor Necrosis Factor alpha enhances α-synuclein accumulation in induced pluripotent stem cell-derived enteric neurons and glia, and impairs the malate-aspartate shuttle, a key pathway for mitochondrial energy production. This drives a metabolic shift toward glutamine oxidation in patient cells. This metabolic impairment reduces overall mitochondrial function, which is partially rescued by the neuroprotective compound Chicago-Sky-Blue 6B. Furthermore, transcriptomic and histological analyses of human gut tissue from inflammatory bowel disease patients reveal that inflammation-associated metabolic suppression and α-synuclein upregulation occur beyond PD, representing general hallmarks of intestinal inflammation. These findings highlight a conserved metabolic vulnerability in the ENS and establish patient-derived enteric lineages as a robust platform to model inflammatory ENS pathology.

Autism spectrum disorder (ASD) is a complex neurodevelopmental condition with limited treatment options, where neuroinflammation and gut microbiota dysbiosis are emerging as interconnected therapeutic targets. This study evaluated the therapeutic potential of non-gene-edited human chemically induced pluripotent stem cell-derived neural stem cells (hCiPSC-NSCs) in a prenatal valproic acid (VPA)-induced rat model of ASD, using a dual-pathway administration strategy (intravenous systemic delivery combined with an intracerebroventricular boost). The treatment significantly ameliorated core ASD-like behaviors, including improved sociability (increased stranger interaction time, P < 0.0001), reduced repetitive behaviors (decreased marble-burying, P < 0.0001; and self-grooming, P < 0.05), and enhanced spatial memory (shorter escape latency in the Morris water maze, P < 0.01). At the mechanistic level, hCiPSC-NSCs attenuated neuroinflammation (suppressed IL-1β, IL-6, and TNF-α; elevated IL-10, all P < 0.0001), reduced oxidative stress (restored GSH and SOD, decreased MDA and NO), diminished microglial activation in the hippocampus and cortex, and restored synaptic ultrastructure by replenishing synaptic vesicles. Furthermore, 16S rRNA sequencing revealed a rebalancing of the gut microbiota, characterized by a reduced Firmicutes/Bacteroidota ratio, enrichment of beneficial taxa like Bacteroidota and Alloprevotella, suppression of pathobionts such as Desulfovibrionales, and partial restoration of microbial diversity. These findings demonstrate that non-gene-edited hCiPSC-NSCs can simultaneously address neural pathophysiology and gut ecosystem disruption in ASD, highlighting their potential as a gut-brain axis-targeting therapy for neurodevelopmental disorders.

Acute myeloid leukemia (AML) remains a highly heterogeneous hematologic malignancy in which therapeutic resistance and disease relapse are largely driven by leukemic stem cells (LSCs). These rare, self-renewing cells possess unique biological properties that enable them to survive conventional chemotherapy and targeted therapies, thereby sustaining minimal residual disease and promoting leukemia re-emergence. LSC persistence arises from a complex and multilayered network of resistance mechanisms, including intrinsic cellular programs, adaptive molecular plasticity, and protective interactions within the bone marrow microenvironment. Intrinsic mechanisms include cellular quiescence, enhanced multidrug efflux activity, resistance to apoptosis and senescence, and activation of stress-adaptive pathways such as autophagy. In addition, LSCs exhibit remarkable metabolic and epigenetic flexibility, allowing them to rewire signaling pathways and survive therapeutic pressure. Extrinsic cues from the bone marrow niche, including stromal interactions, cytokine signaling, and metabolic support, further reinforce the survival and drug tolerance of LSCs. Together, these interconnected mechanisms create a highly resilient cellular state that limits the efficacy of current therapies. In this review, we summarize the major biological pathways that sustain LSC-mediated resistance in AML and discuss emerging therapeutic strategies aimed at selectively targeting these cells. A deeper understanding of LSC biology will be critical for the development of combination therapies capable of eradicating minimal residual disease and achieving durable remission in patients with AML.