Cytokinetic abscission genes are linked to cancers and developmental disorders, but the consequences of disrupted abscission in vivo remain under-explored. Previously we showed that in the forebrain of Cep55 knockout (KO) mouse embryos, a subset of neuroepithelial stem cells (NSCs) fail abscission and become binucleate, and some of those undergo p53-mediated apoptosis. Here we use the Cep55 KO to investigate how stochastic abscission failures in a polarized epithelium affect the epithelial architecture. We find that NSCs in Cep55 KO neuroepithelium have preserved epithelial polarity and integrity. However, they have enlarged apical membranes (called apical endfeet), longer primary cilia, and increased biciliation. We then test whether the enlarged apical endfeet arise from filling the space of apoptotic neighbors. Remarkably, blocking apoptosis does not rescue but exacerbates the phenotypes: extra-large apical endfeet have further increased multiciliation, supernumerary centrosomes, and abnormal or multiple nuclei, although epithelial polarity is maintained. These findings elucidate the importance of proper abscission in maintaining polarized epithelial structure, and reveal that p53-mediated apoptosis is a crucial guardian of tissue architecture when cell division defects arise during development and disease.

Nucleotides and coenzymes play critical roles in energy metabolism and cellular signaling and as building blocks of nucleic acids. This work addresses the challenges in the measurement of the phosphorylated metabolites using hydrophilic interaction liquid chromatography coupled with mass spectrometry, which facilitates the separation and detection of polar metabolites. Here, we present optimized HILIC-MS/MS methods for rapid analysis of polar metabolites including nucleotides and their derivatives in complex biological matrices, such as murine adipose, skeletal, and liver tissues, human plasma, and bacteria. The developed methodologies enable separation of key nucleotides and other phosphorylated metabolites within 6 min and cofactors such as NAD+, NADH, NADP+, and NADPH within 4 min. Validation of these methods demonstrated high accuracy, precision, and sensitivity and stresses the substantial impact of matrix effects. The applicability of the methods was also tested on 13C-labeling experiments with mouse pluripotent stem cells. Additionally, sample pretreatment techniques, such as liquid-liquid extraction and solid-phase extraction, were evaluated as a tool to decrease the negative impact of matrix effects in complex samples. This work enhances the analytical capabilities for nucleotide quantification in metabolomics, facilitating the study of metabolic pathways and disease markers.

Stress urinary incontinence (SUI) adversely impacts millions worldwide due to weakened pelvic floor muscles and urethral sphincter dysfunction. To date, there is a lack of effective non-surgical treatment for SUI, and no clear consensus has been reached on the optimal stem cell source under regenerative therapy. Existing studies have shown no precise molecular mechanisms underlying stem cell-mediated external urethral sphincter (EUS) regeneration. Therefore, we investigated the regenerative and reparative potential of our clinical-grade human amniotic fluid stem cells (hAFSCs) for treating SUI.

Hepatitis C virus (HCV) infection is a major cause of hepatocellular carcinoma (HCC). However, how HCV promotes hepatic progenitor cell (HPC) differentiation into hepatic cancer stem cells (HCSCs) remains unclear, particularly the role of β-catenin signaling and epithelial cell adhesion molecule (EpCAM) regulation in this process. We used mouse HPCs (HP14.5 and HP14.5-Core) and human HCC cell lines (Huh7, HepG2) to investigate HCV core function. Cells were transduced with AdCore, AdGFP, or shRNAs targeting EpCAM or β-catenin. Functional properties were evaluated using spheroid formation, ICG uptake, colony formation, migration, invasion, and in vivo tumorigenesis assays. EpCAM promoter activity and Wnt/β-catenin signaling were examined by luciferase reporter assays. Protein expression was measured by RT-PCR and Western blotting, while immunofluorescence and co-immunoprecipitation were used to analyze HCV core-β-catenin interactions. HCV core expression increased spheroid number, reduced ICG uptake, and accelerated tumor growth in xenograft and orthotopic mouse models, consistent with HPC differentiation toward HCSCs. EpCAM, CD133, CD44, and CD90 were strongly upregulated, whereas EpCAM silencing suppressed proliferation, migration, and invasion. Reporter assays revealed that HCV core enhanced EpCAM promoter activity and activated Wnt/β-catenin signaling. Immunofluorescence demonstrated β-catenin nuclear translocation, and co-immunoprecipitation confirmed direct interaction between HCV core and β-catenin. Knockdown of β-catenin reduced EpCAM, c-Myc, and cyclin D1 expression, diminished spheroid formation, restored ICG uptake, and significantly impaired tumorigenesis in vivo. The HCV core-β-catenin-EpCAM axis drives the conversion of HPCs into HCSCs. This pathway is a central mechanism in HCV-related HCC.IMPORTANCEHCV core protein directly interacts with β-catenin, driving its nuclear translocation and activating EpCAM expression. This promotes HPC differentiation into HCSCs, marked by increased spheroid formation, migration, invasion, and tumorigenesis. Silencing EpCAM or β-catenin reverses these effects, confirming that the HCV core-β-catenin-EpCAM axis is a key pathway in HCV-related HCC.

One nucleotide substitution in codon -18 of HLA-B*40:01:02:01 results in a novel null allele, HLA-B*40:636N.

The identification of cell types remains a major challenge. Even after a decade of single-cell RNA sequencing (scRNA-seq), reasonable cell type annotations almost always include manual non-automated steps. The identification of orthologous cell types across species complicates matters even more, but at the same time strengthens the confidence in the assignment. Here, we generate and analyze a dataset consisting of embryoid bodies (EBs) derived from induced pluripotent stem cells (iPSCs) of four primate species: humans, orangutans, cynomolgus, and rhesus macaques. This kind of data includes a continuum of developmental cell types, multiple batch effects (i.e. species and individuals) and uneven cell type compositions and hence poses many challenges. We developed a semi-automated computational pipeline combining classification and marker-based cluster annotation to identify orthologous cell types across primates. This approach enabled the investigation of cross-species conservation of gene expression. Consistent with previous studies, our data confirm that broadly expressed genes are more conserved than cell type-specific genes, raising the question of how conserved, inherently cell type-specific, marker genes are. Our analyses reveal that human marker genes are less effective in macaques and vice versa, highlighting the limited transferability of markers across species. Overall, our study advances the identification of orthologous cell types across species, provides a well-curated cell type reference for future in vitro studies and informs the transferability of marker genes across species.

Alzheimer's disease (AD) exhibits high genetic and clinical heterogeneity that limits therapeutic success. Patient-derived brain organoids and their extracellular vesicles (EVs) provide physiologically relevant models to study disease mechanisms and individualized drug responses.

The developing embryo harbors multiple hematopoietic programs, categorized as either intra-embryonic or extra-embryonic yolk-sac, that differ in their spatio-temporal origins and developmental potential. In the vertebrate embryo, the hematopoietic stem cell (HSC) derives from the definitive intra-embryonic hematopoietic program and is dependent on stage-specific retinoic acid (RA) signaling. We have recently modelled aspects of this developmental process in vitro using human pluripotent stem cells (hPSCs) and identified a KDR+CXCR4+ mesodermal population that generates definitive hematopoietic progeny in a uniquely RA-dependent manner. A subpopulation of this mesoderm expresses ALDH1A2, an enzyme involved in RA synthesis. Here, we sought to characterize the role of ALDH1A2 in the development of the human RA-dependent hematopoietic lineage and to map its mesodermal origin. Using two different engineered reporter hPSC lines, we show that specification of this lineage requires a functional ALDH1A2 enzyme at the mesoderm stage. Through functional analyses of different mesoderm subpopulations, we demonstrate that this RA-dependent lineage derives from ALDH1A2neg mesoderm by non-cell autonomous RA signaling. Collectively, these studies provide new insight into the differentiation trajectory of hPSCs towards the definitive hematopoietic lineage.

Short QT syndrome is a heritable arrhythmia disorder linked to sudden cardiac death. We recently identified that individuals with alternating hemiplegia of childhood (AHC), a rare neurodevelopmental disorder, can exhibit shortened corrected QT intervals and elevated risk for ventricular fibrillation. This is especially true for patients with AHC heterozygous for the recurrent ATP1A3-D801N variant, though the underlying cardiac mechanism remains unclear. We hypothesized that the D801N missense impairs Na+/K+-ATPase function, causing Ca2+ overload, shortened action potential duration (APD), and arrhythmias. Using in silico modeling and patient-derived induced pluripotent stem cell cardiomyocytes (iPSC-CMsD801N), we observed shorter APD, elevated intracellular and sarcoplasmic reticulum Ca2+ levels, and delayed afterdepolarizations (DADs) compared with WT. Additionally, increased Ca²+ influx via the Na+/Ca2+ exchanger (NCX1) during depolarization was observed in iPSC-CMsD801N. Simulations and in vitro experiments suggest that reduced ATPase function accelerated inactivation of L-type Ca2+ channels. Pharmacologic inhibition of NCX1 with ORM-10103 normalized APD and reduced DADs. These findings support a Ca2+-mediated mechanism for arrhythmogenesis in ATP1A3-D801N carriers and identify NCX1 as a potential therapeutic target.

Bronchopulmonary dysplasia (BPD) is a common and serious complication among preterm infants, particularly those born at extremely low gestational ages. It is primarily characterized by impaired alveolar and vascular development. Inflammation is increasingly recognized as a central mechanism in its pathogenesis. Both prenatal factors, such as intrauterine infection, and postnatal insults, including mechanical ventilation, oxygen toxicity, and infection, can trigger and sustain a dysregulated inflammatory response in the immature lung. This response involves the activation of inflammatory cells, such as neutrophils and macrophages, and the release of pro-inflammatory mediators, reactive oxygen species (ROS), and proteases. These factors disrupt critical developmental signaling pathways and contribute to alveolar simplification and abnormal vascular growth, which are the hallmark features of BPD. Current therapeutic strategies aim to limit these inflammatory processes and support lung development. Established interventions like caffeine and corticosteroids have demonstrated varying levels of effectiveness and safety. Emerging therapies-including anti-cytokine agents, inflammasome inhibitors, and stem cell-based approaches-offer promising avenues by specifically targeting the inflammatory cascade. Additionally, supportive strategies such as non-invasive ventilation, careful oxygen titration, and optimal nutrition play essential roles in reducing initial injury and facilitating recovery. Inflammation is a key mediator linking diverse perinatal insults to the disrupted lung development seen in BPD. A deeper understanding of the inflammatory mechanisms and timely, targeted interventions may offer improved outcomes for this vulnerable population.

Neural progenitor cells (NPCs) are promising candidates for cell replacement therapies, yet maintaining stemness while enabling expansion in chemically defined three-dimensional (3D) hydrogels remains a challenge. By tuning crosslink exchange kinetics, crosslinker functionality and stoichiometry, polymer phase separation behavior, and adhesive ligand presentation, a family of hydrogels was prepared to study the effects of stress relaxation timescale and network connectivity on NPC phenotype. Hydrogels with rapid relaxation and low connectivity promote expansion of NPCs as distributed single-cell networks that maintain stemness marker expression and differentiation capacity. NPCs embedded in slowly relaxing hydrogels maintained stemness marker expression through cell clustering but exhibited impaired proliferation and differentiation. Similarly, in the absence of integrin-binding cell adhesive ligands, NPCs also maintained stem cell marker expression but remained as clusters rather than distributed single-cell networks. Cadherin cell-cell contacts enable downstream β-catenin signaling and stemness maintenance, which are enhanced in rapidly relaxing, low connectivity networks. These findings identify a combination of network connectivity, stress relaxation timescale, and integrin-binding adhesive ligands as crucial design parameters for maintaining NPC stemness and differentiation capacity in 3D hydrogel networks.

Tyrosine kinase inhibitors (TKIs) targeting ABL1 catalytic activity have markedly improved Chronic Myeloid Leukemia (CML) outcomes, inducing unprecedented and durable therapeutic responses. However, while TKIs efficiently target committed leukemic progenitors, they fail to eradicate leukemic stem cells (LSCs), which may drive disease relapse. High BCR::ABL1 transcripts at diagnosis confer a proliferative and survival advantage and are associated with a higher risk of CML progression to the acute phase. Specifically Targeting the ABL Myristoyl Pocket (STAMP) compounds, including asciminib (ASC), provide a novel mechanism to inhibit BCR::ABL1 catalytic activity. ASC is FDA-approved for patients who have failed one or more TKIs, and its efficacy has been evaluated as monotherapy, and in combination with different TKIs, in T315I-positive or advanced-phase CML. We investigated the cytotoxic effects of ASC, alone or with imatinib (IM) or nilotinib (NIL), on committed progenitors and LSCs from CML patients expressing high or low BCR::ABL1 at diagnosis. ASC reduced BCR::ABL1-dependent survival and impaired clonogenicity in committed progenitors with low, but not high, BCR::ABL1 transcripts. ASC also disrupted LSC self-renewal, reducing both frequency and number of Long-Term Culture-Initiating Cells (LTC-ICs). When combined with IM or NIL, ASC restored TKI activity against LTC-ICs expressing high BCR::ABL1 transcripts, with the association of ASC and NIL reducing both LTC-IC division rates and LTC-IC-derived CFUs. These findings suggest that ASC, alone or with NIL, may target LSCs and improve outcomes in patients with high BCR::ABL1 expression at diagnosis.

The limited regenerative capacity of cartilage tissue and the high morbidity associated with injuries and diseases have driven the search for innovative regenerative medicine strategies. The objective of the study was to compare the chondrogenic differentiation of human MSCs in conventional pellet cultures to that of spheroids generated using an innovative microwell system.

Extracellular vesicles (EVs) are potential cell-free therapies for cardiac regeneration. Although adipose-derived stem cells (ADSCs) are easily obtained using minimally invasive procedures, therapeutic effects of hypoxically conditioned ADSC-derived EVs on the heart remain unknown. We aimed to verify whether hypoxic preconditioning enhances the therapeutic efficacy of ADSC-derived EVs.

Osteoarthritis (OA) is the most prevalent chronic degenerative joint disorder worldwide, characterized by progressive cartilage degradation, subchondral bone remodeling, synovial inflammation, and impaired mobility. Growing evidence has established mitochondrial dysfunction-including impaired oxidative phosphorylation (OXPHOS), excessive reactive oxygen species (ROS) generation, disrupted mitochondrial dynamics, and dysregulated mitophagy-as an early and pivotal driver of OA pathogenesis. These bioenergetic failures not only disrupt chondrocyte metabolism but also amplify inflammation, matrix degradation, and cell death. In recent years, mitochondrial transplantation has emerged as a revolutionary therapeutic paradigm, aiming to restore cellular homeostasis by delivering functional mitochondria into damaged chondrocytes. This review systematically summarizes the molecular mechanisms of mitochondrial dysfunction in OA and highlights three major therapeutic strategies: (1) cell-based approaches, particularly mesenchymal stem cell (MSC)-mediated mitochondrial transfer via tunneling nanotubes (TNTs) or extracellular vesicles (EVs); (2) cell-free approaches, utilizing purified mitochondria or MitoEVs for direct transplantation; and (3) engineered mitochondrial transplantation, integrating bioengineering, nanotechnology, and genetic modification to enhance mitochondrial quality, delivery efficiency, and therapeutic persistence. We further discuss opportunities and challenges in clinical translation, including standardization of mitochondrial preparation, optimization of delivery systems, immunological safety, and regulatory classification. Collectively, mitochondrial transplantation represents a disruptive strategy that directly addresses the bioenergetic collapse of chondrocytes and offers a promising avenue for disease-modifying therapy in OA. Future advances in mechanistic elucidation, technological optimization, and multicenter clinical trials will be crucial to transform "mitochondrial medicine" from experimental concept to clinical reality.

The management of bronchial asthma has evolved from a one-size-fits-all approach to the era of precision medicine, which is guided by intrinsic phenotypes. This article systematically reviews the mechanisms of action of current targeted biologics (targeting pathways such as IgE, IL-5/IL-5R, IL-4/IL-13, and TSLP), the efficacy and safety data derived from pivotal clinical trials, and the biomarker systems that guide clinical decision-making, further elaborating on how to implement tailored individualized therapy based on patient-specific characteristics. However, existing biologics still face challenges including the need for long-term administration and the inability to reverse disease progression. Therefore, this article focuses on the transformative prospects of next-generation therapeutic modalities. Cell therapy represents the most promising breakthrough, with its core shifting from "passive suppression" to "active regulation and remodeling". This is mainly reflected in three cutting-edge areas: cellular reprogramming (e.g., converting pathogenic Th2 cells into homing-competent regulatory T cells), engineering modification (e.g., designing CAR-NK cells with dual functions of targeted clearance and immune regulation), and multifunctional immune/repair modulation (e.g., utilizing mesenchymal stem cells and their exosomes to suppress immune abnormalities at the source and promote tissue repair). Collectively, these strategies drive a fundamental shift in treatment goals from symptom control to the induction of long-term immune tolerance and even functional cure. In conclusion, the future management of asthma will be a dynamically evolving individualized integrated system. By deeply integrating targeted biologics, intelligent advanced cell therapies, and continuously optimized precision management strategies, we are expected to ultimately establish a multi-level, closed-loop diagnosis and treatment pathway for each patient, laying the foundation for achieving long-term high-quality remission.

Postmenopausal osteoporosis (PMO) develops as a result of pathological cross-tissue interactions. However, current experimental paradigms are constrained by their single-tissue focus, hindering efforts to discover systemwide regulatory genes.

Due to toxicity, high costs, and potential side effects of standard treatments of rheumatoid arthritis (RA) including nonsteroidal anti-inflammatory drugs (NSAIDs), corticosteroids, and disease-modifying antirheumatic drugs (DMARDs), natural products and advanced drug delivery systems, such as nanoparticles and mesenchymal stem cell (MSC)-derived exosomes (EXO), have garnered interest due to their ability to target inflammation and oxidative damage, with enhanced precision and reduced side effects, offering a promising approach for RA management.

Epithelial stem cells in the interfollicular epidermis (IFE) and hair follicle (HF) play key roles in maintaining and regenerating the skin barrier by balancing self-renewal and differentiation. These fate decisions are governed by transcriptional and epigenetic mechanisms that respond to context-dependent signals from the skin microenvironment. In the IFE, basal stem cell divisions follow a stable pattern but rely on tightly regulated transcription factors and chromatin states to ensure proper epidermal maintenance. In contrast, HF stem cells exhibit a higher degree of plasticity that allows for rapid adaptation to changing environments, including IFE regeneration following injury. While this plasticity is critical for epidermal integrity, it can also drive disease onset if transcriptional programs become disrupted. This review provides a comprehensive analysis of how transcriptional and epigenetic regulators guide stem cell fate decisions required in the IFE and HF that promote epidermal homeostasis. We also explore how these programs are altered in various pathological contexts in the skin. By comparing differentiation mechanisms in the IFE and HF compartments, we highlight how dynamic control of gene expression sustains skin homeostasis.

Regeneration of tubular dentine structure is key to its biological function, and the polarity of odontoblasts is crucial for this, but the mechanism is unclear. On the basis of differential gene expression data comparing odontoblasts and donor-matched osteoblasts, we hypothesized that forkhead box Q1 (Foxq1), a key regulator in embryonic development, plays a significant role in the differentiation of polarized odontoblasts. This study aimed to investigate the role of Foxq1 in odontoblast polarization and tubular-dentine formation, and to explore its relationship with Wingless-type family member 5A (Wnt5a) signalling.