Not available.

Single-cell passaging of induced pluripotent stem cells (iPSCs) has historically been discredited, as it may risk mutations. Here, however, in combination with Rho kinase inhibition and based on stringent quality control, we successfully validate a corresponding iPSC banking process under Good Manufacturing Practice (GMP) conditions. The underlying culture system features enhanced consistency, sustains genomic integrity in the resulting cell lines, and enables excellent priming for gene editing and directed differentiation. The process, as well as the resulting GMP iPSC lines, will provide a robust foundation for clinical manufacturing.

Globally, third leading cause of cancer-related deaths is contributed by Hepatocellular carcinoma (HCC). Chronic hepatitis B virus infection is one of the seminal etiological drivers of HCC. Hepatitis B viral DNA integration, host genomic instability, persistent inflammatory responses and the oncogenic activity of the viral oncoprotein Hepatitis B virus X (HBx), contribute to the hepatocarcinogenesis. Emerging evidences indicate that epigenetic dysregulation plays a seminal role in linking viral persistence in the liver tissue to its malignant transformation. In HBV-infected hepatocytes, aberrant DNA methylation, histone modifications, and dysregulated non-coding RNAs reprogram transcriptional networks that activate oncogenic pathways, promote proliferative signaling, and sustain cancer stem cell-like phenotypes driving HCC progression. The epigenetic modifications in the infected, malignant hepatic cells can influence the tumor microenvironment, contributing to the infiltration of exhausted cytotoxic T lymphocytes with elevated PD-1 and Tim-3 expression. Further, the T lymphocytes exhibit reduced proliferative capacity, impaired cytokine secretion, and diminished cytotoxic activity. In the clinical perspective, long-term nucleotide analogue therapy causes viral suppression and attenuation of inflammation, thereby reducing HCC progression by 40-80%. Despite the extensive T-cell exhaustion, HBV-associated HCC (HBV-HCC) is responsive to immune checkpoint blockade, as highlighted in the CheckMate-040 trial. Emerging therapeutic strategies combine anti-viral agents with immune checkpoint inhibitors, epi-drugs and HBsAg-directed TCR-engineered T cells. These clinical approaches aim to simultaneously restore antitumor immune responses as well as neutralize the viral oncogenic drivers, offering promising avenues for improved management of HBV-induced HCC.

Employing a comprehensive strategy that integrated network pharmacology, molecular docking, and in vitro experimental techniques, this study examined the effect of neohesperidin on the proliferation and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) upon lipopolysaccharide (LPS)-induced inflammatory conditions.

Volumetric Musculoskeletal Trauma (VMST), which is characterized by volumetric muscle loss accompanied by bone injury, poses a significant challenge to regenerative medicine. While current therapies primarily focus on the individual regeneration of muscle or bone, there is no systematic and integrated treatment strategy. In this study, we developed a CuMBG-PA, a copper-doped nanozyme based on mesoporous bioactive glass (MBG) and procyanidin (PA), for integrated muscle and bone repair of VMST. CuMBG-PA self-assembles into a stable polyphenol network via Cu-PA coordination bonds, enhancing PA stability and reactive oxygen species-scavenging capacity. In vitro and in vivo experiments demonstrated that CuMBG-PA promoted osteogenesis and myogenesis while exhibiting high biocompatibility and antibacterial activity. Single-cell RNA-sequencing results revealed that CuMBG-PA synergistically induces stem cell differentiation and promotes tissue repair through multiple myoskeletal shared metabolic pathways. Mechanistically, CuMBG-PA exerts its beneficial effects by increasing the number of Proteoglycan 4 (Prg4) + fibro/adipogenic progenitor cells (FAPs), which highly express fibronectin and insulin-like growth factor. In addition, PRG4 regulates immune cells, removes overactivated muscle satellite cells, reduces muscle fibrosis, and promotes functional recovery during regeneration. In summary, this work demonstrates that the novel self-assembled CuMBG-PA nanozyme represents a potential biomaterial-based strategy for integrated muscle and bone repair in VMST.

Patients with polycystic ovary syndrome (PCOS) are at an elevated risk of depression, yet the underlying mechanisms remain elusive. Emerging evidence implicates the gut-brain axis and systemic lipid homeostasis alterations as potential key contributors. We profiled untargeted plasma lipidomes of PCOS patients with and without comorbid depression (PCOS-DP) and integrated these data with our prior gut microbial and host transcriptomic datasets to construct multi-omics interaction networks. The causal role of the candidate gut microbial was preliminary explored in a germ-free PCOS mouse model using fecal microbiota transplantation, followed by behavioral phenotyping and ELISA-based protein quantification. We identified a distinct plasma lipidomic signature differentiating PCOS-DP from PCOS alone, characterized primarily by the downregulation of 26 lipid species. Most of these altered lipids were triacylglycerols (TAGs) enriched with FA18:1 and FA18:2, whose levels correlated with coagulation dysfunction. Multi-omics network analysis revealed significant interconnections between depression-associated gut microbiota (including Bacteroides eggerthii), specific altered lipids such as TAG (60:12/FA22:6), and host genes involved in inflammation (e.g., IL22, NLRP7), metabolism, and neural processes. Animal validation demonstrated that B. eggerthii colonization in PCOS mice specifically exacerbated anhedonia and hyperlocomotion, alongside modulating plasma IL-22 expression, suggesting its context-dependent neurobehavioral effect role. This study delineates a TAG-downregulated lipid signature with diagnostic potential and reveals a novel "gut microbiota-lipid-host gene" interaction network underpinning PCOS-DP, with B. eggerthii as a key microbial modulator of neurobehavioral phenotypes in the context of PCOS. These findings provide new pathophysiological insights and highlights potential diagnostic biomarkers for PCOS-DP.

Repairing large-scale craniomaxillofacial bone defects is hindered by a limited availability of stem-cell sources and a low osteogenic efficiency. To address these challenges, Fe3O4 nanoparticles were modified with methacrylic anhydride (MAA), which helped to introduce photopolymerizable methacryloyl groups, resulting in MAA-Fe3O4 nanoparticles that exhibit excellent magnetic properties and colloidal stability. These nanoparticles were incorporated into gelatin methacryloyl (GelMA) and covalently crosslinked to form an injectable, photocurable GelMA-Fe3O4 magnetic composite hydrogel. This hydrogel provided a three-dimensional culture microenvironment for human dental follicle stem cells (hDFSCs), and upon encapsulation, osteogenesis was significantly enhanced under a 100 mT static magnetic field (SMF). In vitro, GelMA-Fe3O4 hydrogels demonstrated increased porosity and improved mechanical properties, thereby significantly promoting hDFSCs proliferation, adhesion and spreading. Additionally, under SMF exposure, the expression of osteogenesis-related genes and proteins, including alkaline phosphatase (ALP), Runx2, Col-I and OPN, was significantly upregulated. In a rat calvarial defect model, bone mineralization centers with multi-site distribution were observed in the GelMA-Fe3O4 + SMF group as early as 4 weeks postoperatively, leading to high-quality defect repair. The limitations of traditional 'peripheral-to-center' unidirectional repair were overcome by this model of synchronous multi-site osteogenesis, maximizing bone regeneration with a minimal number of stem cells and providing an efficient, controllable tissue-engineering strategy for the clinical treatment of craniomaxillofacial bone defects.

Volumetric Muscle Loss presents a critical challenge involving the traumatic or surgical loss of over 20% of skeletal muscle mass by overwhelming the body's natural regenerative capacity. It causes functional decline of skeletal muscles leading to reduced quality of life. Current surgical interventions, such as autograft and allograft muscle transfers, often fall short of restoring full mobility frequently causing donor site morbidity and graft failure. The objective of this manuscript is to discuss the role of emerging regenerative strategies focusing on restoring muscle structure and regenerative microenvironment. Recent advances emphasize on extracellular matrix-based therapies that promote myogenesis and vascularization because of their ability to replicate the native structural as well as biochemical attributes leading to muscle fiber regeneration and innervation. Further, incorporation of growth factors like vascular endothelial growth factor (VEGF), insulin-like growth factor-1 (IGF-1), or stem cells in the scaffolds help to recapitulate the complex structure and signaling of extracellular matrix promoting accelerated healing and recovery as observed in pre-clinical trials. However, despite of positive outcomes, there are challenges like immunogenicity, issues with batch to batch reproducibility, which hinder scalability and translation. Interdisciplinary collaboration between biomaterials science, tissue engineering, and clinical research can serve as solution to resolve this critical issue and will be helpful to advance these technologies potentially shifting the approach of VML therapeutic management from palliative to curative.

Adult zebrafish exhibit full recovery following spinal cord injury. Transient expansion of stem cell-like progenitors is thought to underlie their regenerative capacity. Yet, our understanding of the identities and contributions of the crucial stem cell populations that direct spontaneous neural repair remains limited. Moreover, while most neural regeneration research is centered on promoting proliferative repair, the regulatory mechanisms that reinstate quiescence post-repair are unknown. Here, we determined the molecular identities and cellular contributions of sox2+ progenitors during spinal cord repair. Genetic lineage tracing shows zebrafish spinal progenitors, while quiescent in uninjured tissues, self-renew and differentiate into neurons and glia after injury. By single-cell sequencing, sox2 + cells are heterogeneous and biased towards neuronal or glial fates in both homeostatic and regenerating tissues. By screening for transcription factors that are differentially expressed in acute versus chronic spinal cord injury, we find the Bach1 transcription factors control transient progenitor cell activation by acting as dual activators and repressors of sox2 expression. This study elucidates the molecular diversity and contributions of sox2 expressing cells during spinal cord repair and identifies a transcriptional regulatory switch by which progenitor cells expand after injury and restore quiescence after regeneration is completed.

Recent genetic discoveries in congenital hyperinsulinism (HI) and advances in sequencing technology suggest that the diagnostic yield may be improved by rescreening in people with genetically unsolved HI.

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.

Neurological injuries and neurodegenerative disorders, including spinal cord injury, traumatic brain injury, stroke, and Parkinson's disease remain largely incurable. In the central nervous system (CNS), a self-reinforcing cascade of neuroinflammation, oxidative stress, blood-brain barrier breakdown, and glial fibrotic scarring restricts long-distance axonal regrowth and graft survival. The peripheral nervous system (PNS) exhibits greater intrinsic regenerative potential, yet critical-length defects remain challenging and have driven the development of clinically relevant conduit designs. This review provides an overview of the microenvironment following CNS injury and summarizes the key design requirements for engineered repair matrices, while highlighting lessons from advanced peripheral nerve guidance conduits. Injectable extracellular matrix (ECM)-mimetic and smart hydrogels can conformally fill CNS cavities, modulate immune and redox cascades, restore vascular function, and provide permissive niches for neural stem/progenitor and endothelial cells. CNS-compatible bioinks and 3D bioprinting enable the fabrication of neurovascular architectures and multicellular constructs with controlled mechanics, topology, and circuit geometry. Advances in nerve guidance conduits inform translation of PNS principles to the brain and spinal cord. Organoid-based strategies, including vascularized organoids, biomaterial-supported grafts, and organoid-neuroelectronic interfaces, suggest routes toward modular biohybrid constructs. Integrating pathology-informed biomaterials, biofabrication, and organoid engineering offers a roadmap for neural circuit reconstruction.

Despite extensive global efforts to eradicate cervical cancer through improved screening and widespread HPV vaccination, the disease continues to claim over 350 000 lives annually. Persistent infection with high-risk HPV genotypes, primarily HPV-16 and HPV-18, is the principal etiological driver, with viral DNA integration into the host genome representing a critical event in malignant progression. In this study, HPV integration was investigated using 25 RNA-seq datasets from HPV-positive cervical cancer cell lines and 2 datasets from the HPV-negative C-33A cell line. Analysis revealed a recurrent HPV-16 integration hotspot on Chromosome 6 within the CLIC5 gene. This integration was validated by qRT-PCR and Sanger sequencing. Subsequent gene expression analysis demonstrated significant CLIC5 upregulation in HPV-16-positive cell lines, with a 21-fold increase in CaSki and a 4-fold increase in SiHa compared to HPV-negative and HPV-18-positive controls. These findings delineate a distinct HPV-16 integration pattern and its associated transcriptional impact. The identification and validation of this strain-specific integration site nominates HPV-16 integration at the CLIC5 locus as a potential biomarker for HPV-driven cervical cancer.

To develop novel hydropyrimidine linked amide-pyridine hybrids as anticancer agents.

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.