Angelman syndrome (AS) is a rare neurodevelopmental disorder characterized by severe developmental delay, speech impairment, ataxia and happy demeanor. AS is caused by loss-of-function of maternal UBE3A in neurons due to (epi)genetic abnormalities. Here, we report two new induced pluripotent stem cell (iPSC) lines from male and female patients carrying ∼ 6 Mb deletions in chr15q11.2-q13.1, together with familial control iPSC lines. All lines express pluripotent stem cell markers, demonstrate trilineage differentiation, and maintain genetic and epigenetic integrity at the locus of interest. These iPSCs provide a platform to model class I deletions, the most severe AS cause, and accelerate therapy development.

Hematopoietic stem cells (HSC) rapidly expand during fetal development and after stress. Here, we identify BCLAF1 as a regulator of HSC repopulation activity with functions in expansion of fetal HSCs and hematopoietic reconstitution after stem cell transplantation. Using mice with hematopoietic-specific and inducible deletion of Bclaf1, we find that BCLAF1 promotes fetal HSC development but is dispensible for maintenance of adult HSCs at steady state. Loss of BCLAF1 in either fetal or adult HSCs significantly impairs their self-renewal and multi-lineage reconstitution activity after stem cell transplantation. Single-cell RNA sequencing of fetal hematopoietic progenitors reveals that loss of BCLAF1 reduces long-term HSCs and restrains expression of stress response genes. BCLAF1 associates with chromatin throughout the genome of fetal and adult hematopoietic cells, likely through indirect mechanisms, to regulate transcriptional programs. These results establish a novel function for the transcriptional regulator BCLAF1 in limiting stress responses in HSCs to preserve HSC development during embryogenesis and repopulation function after stem cell transplant.

Excessive glucocorticoid use disrupts osteogenesis and angiogenesis in the femoral head, leading to steroid-induced osteonecrosis of the femoral head (SONFH), which is a significant clinical challenge. This study introduces a magnetically responsive biphasic calcium phosphate (HBCP/Fe3O4) scaffold featuring a nanoparticle-embedded hollow-tube whisker structure. The scaffold was fabricated through an in situ growth process to generate hollow-tube whiskers, followed by a capillary trapping technique that allowed the hollow-tube whiskers to capture Fe3O4 nanoparticles (NPs), achieving uniform and efficient encapsulation. HBCP/Fe3O4 exhibited excellent magnetic responsiveness and significant biological effects under static magnetic field (SMF) stimulation. In vitro, HBCP/Fe3O4 under SMF promoted osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in a glucocorticoid microenvironment, enhanced angiogenesis in human umbilical vein endothelial cells (HUVECs), and induced M2 polarization of RAW 264.7 murine macrophage cells (RAW 264.7). Furthermore, HBCP/Fe3O4 under SMF stimulation orchestrated paracrine signaling from endothelial and immune cells, thereby enhancing the osteogenic differentiation of BMSCs. Mechanistically, the osteogenic differentiation of BMSCs was driven by magnetic stimulation-induced Piezo1-mediated Ca2+ influx, which activated BMP-2/Smad signaling and upregulated key osteogenic markers. In vivo, the implantation of HBCP/Fe3O4 scaffolds under SMF stimulation in a rabbit SONFH model promoted coordinated therapeutic effects, including robust bone regeneration, in situ revascularization, immunomodulation, and preservation of femoral head cartilage. Together, these findings support the clinical relevance of this magnetically responsive scaffold as a multifunctional strategy for delaying structural deterioration and facilitating comprehensive repair in SONFH.

To develop effective therapies for glioblastoma (GBM), a deeper understanding of its underlying immunoregulatory mechanisms is needed. Invariant natural killer T (iNKT) cells are unconventional T cells that recognize lipid antigens and are known to regulate tumor immunity in other cancer types. Given the lipid-rich nature of the brain and the unique metabolic activity of GBM cells, we hypothesized that GBM-enriched lipids could direct iNKT cells to contribute to the immunosuppressive nature of the disease.

[This retracts the article DOI: 10.1155/2016/7380439.].

The reversible epitranscriptomic mark, 5-methylcytosine (m5C) modification, is implicated in numerous cellular processes, but its role in neural development remains largely unexplored. In this study, we discovered high expression of the m5C reader Ybx1 in the developing mouse cortex. To elucidate its role in cortical development, Ybx1 was ablated in embryonic cortical neural stem cells (NSCs). Interestingly, conditional knockout (cKO) of Ybx1 led to perinatal mortality in mice, along with abnormal cortical development. Cortical progenitor cells lacking Ybx1 exhibited impaired proliferation and differentiation. Multi-omics analysis identified the target mRNAs of Ybx1, which encode the key cell cycle regulatory proteins converging on cyclin D2 (Ccnd2). Ybx1 was found to regulate the stability of its target transcripts. Both knockdown and overexpression of Ybx1 targets via in utero electroporation confirmed that they mediated Ybx1 regulation of proliferation and differentiation of neural precursor cells. Further analysis showed that the G1 to S phase transition in cortical progenitor cells is delayed in the Ybx1 cKO. This study highlights the crucial function of the m5C reader protein Ybx1 in promoting cell cycle progression of the embryonic cortical progenitors, essential for proper cortical development.

The mechanisms by which epithelial stem cells (SCs) sense mechanical cues within their niche and convert the information into biochemical signals to govern their function are not well understood. Here, we show that hair follicle SCs (HF-SCs) sense mechanical forces through cell adhesion and maintain quiescence in a PIEZO1-dependent mechanism. PIEZO1 interacts with E-cadherin in HF-SCs, and mechanical pulling of E-cadherin with a force of ~20 pN triggers PIEZO1-dependent, localized calcium flickers. Deletion of Piezo1 leads to reduced cumulative calcium influx and compromises quiescence. Single-cell genomic analyses identify a transcriptional network involving AP1 and NFATC1, which functions downstream of PIEZO1 and regulates the expression of extracellular matrix, cell adhesion, and actin cytoskeleton genes to reinforce the unique mechanical property of HF-SCs. These findings establish the force threshold necessary for PIEZO1 activation and reveal PIEZO1-dependent calcium influx as a key mechanism for sensing mechanical cues in the niche and regulating HF-SC activity.

Genomes adapt dynamically to alterations in the signaling milieu, including inflammation that transiently or permanently disrupts genome function. Here, we elucidate how a progenitor cell genome senses and responds to inflammation when the developmental and transcriptional regulator GATA2 is limiting, which causes bone marrow failure in humans and mice and predisposes to leukemia in humans. GATA2low murine progenitors are hypersensitive to inflammatory mediators. We discovered that the hematopoietic transcription factor PU.1 conferred transcriptional activation in GATA2low progenitors in response to Interferon-γ and Toll-Like Receptor 1/2 agonists. In a locus-specific manner, inflammation reconfigured genome activity by promoting PU.1 recruitment to chromatin or tuning activity of PU.1-preoccupied chromatin. The recruitment mechanism disproportionately required IKKβ activity. Inflammation-activated genes were enriched in motifs for RUNX factors that cooperate with GATA factors. Contrasting with the GATA2-RUNX1 cooperativity paradigm, GATA2 suppressed and RUNX1 promoted PU.1 mechanisms to endow the progenitor genome with inflammation-sensing capacity.

Hashimoto's thyroiditis (HT) is a prevalent autoimmune disease without a cure. Mesenchymal stem cells (MSC) may offer the opportunity to improve autoimmune thyroiditis.

Bone morphogenetic protein (BMP) antagonists have been increasingly linked to the development of colorectal cancer (CRC). BMP signalling operates in opposition to the WNT signalling pathway, which sustains stem-cell maintenance and self-renewal of the normal intestinal epithelium. Reduced BMP and elevated WNT signalling lead to expansion of the stem-cell compartment and the hyperproliferation of epithelial cells, a defining characteristic of CRC. Chordin-like-2 (CHRDL2) is a secreted BMP antagonist, with overexpression linked to poor prognosis and variants in the gene shown to be associated with an elevated CRC risk. However, the detailed mechanism by which CHRDL2 contributes to CRC is unknown. In this study, we explored the impact of CHRDL2 overexpression on CRC cells to investigate whether CHRDL2's inhibition of BMP signalling intensifies WNT signalling and enhances the cancer stem-cell phenotype and response to treatment. Our research approach combines 2D cancer cell lines engineered to inducibly overexpress CHRDL2 and 3D organoid models treated with extrinsic CHRDL2, complemented by RNA sequencing analysis. CHRDL2 was found to enhance the survival of organoids and CRC cells during chemotherapy and irradiation treatment due to activation of DNA damage response pathways. Organoids treated with secreted CHRDL2 exhibited elevated levels of stem-cell markers and reduced differentiation, as evidenced by diminished villi budding. RNA-seq analysis revealed that CHRDL2 increased the expression of stem-cell markers, WNT signalling and other well-established cancer-associated pathways through BMP inhibition. These findings collectively suggest that CHRDL2 overexpression could affect response to CRC therapy by enhancing DNA repair and the stem-cell potential of cancer cells, and its role as a biomarker should be further explored.

The midbody (MB), a transient structure formed during cytokinesis, has evolved from a mere structural component to a complex signaling organelle with diverse functions beyond cell division. Recent studies have revealed that jettisoned midbody remnants (MBRs) play crucial roles in intercellular communication, influencing cell fate decisions, particularly in stem cells and cancer. MBRs act as large extracellular vesicles, transferring functional RNA and proteins that modulate cell behavior, including proliferation and cancer progression. The protein KIF23, associated with midbodies, is a pan-cancer marker, underscoring the clinical relevance of MB research. This review highlights the emerging significance of midbodies and MBRs in cancer biology, neurobiology, and regenerative medicine, offering new avenues for diagnostic and therapeutic strategies. By reshaping our understanding of cell division and intercellular communication, these findings open exciting frontiers in cell biology with huge potential for diagnostic and therapeutic applications.

The cellular demand of the hematopoietic system is maintained by a rare pool of tissue-specific, hematopoietic stem cells (HSCs). HSCs are primarily maintained in a quiescent state but can be activated to exit quiescence and undergo self-renewal and differentiation in response to stress. The cytokine Transforming Growth Factor-β (TGF-β) plays an essential role in supporting HSC quiescence and activation, as one of the most potent inhibitors of HSPC growth. Therefore, how TGF-β signaling can be regulated in the context of HSCs is of significant interest as it may uncover novel mechanisms to target HSC activity. Previous studies revealed that the tetraspanin CD82 modulates the long-term HSC population, with CD82 knockout (KO) mice displaying increased HSC activation. Here, in this study, we connect the CD82 scaffold with the regulation of TGF-β signaling in hematopoietic stem and progenitor cells (HSPCs). We show that CD82KO leads to decreased TGF-β signaling, whereas increased CD82 expression promotes TGF-β activation. These changes in CD82-mediated TGF-β signaling are associated with extracellular matrix interactions, as fibronectin engagement is critical for promoting TGF-β signaling. Mechanistically, we find that CD82 stimulates enhanced TGF-β activation by promoting receptor crosstalk between TGF-β receptor I and integrin β1, resulting in downstream changes in cell proliferation. Collectively, these findings demonstrate that CD82 modulates canonical TGF-β signaling through receptor crosstalk mechanisms that may be targeted to alter the balance between HSC quiescence and activation.

Cardiovascular diseases (CVDs) remain the leading cause of global mortality, with myocardial infarction (MI) and subsequent heart failure (HF) posing significant clinical challenges. Despite advancements in pharmacological and surgical interventions, the limited regenerative capacity of the adult human heart necessitates innovative therapeutic strategies. Stem cell-based therapies have emerged as a promising approach to cardiac regeneration, aiming to restore damaged myocardial tissue through cell replacement and paracrine-mediated repair mechanisms. This review provides a comprehensive overview of the current landscape of stem cell therapies for cardiac regeneration, focusing on the molecular mechanisms, cell types, delivery techniques, and recent clinical advancements. We highlight the roles of key signaling pathways, including NOTCH, PI3K/Akt, Wnt/β-catenin, Hippo/YAP, and MAPK, in regulating cardiomyocyte proliferation, angiogenesis, fibrosis, and inflammation. Additionally, we discuss the therapeutic potential of various stem cell types, such as mesenchymal stem cells (MSCs), cardiac progenitor cells (CPCs), induced pluripotent stem cells (iPSCs), and embryonic stem cells (ESCs), in promoting cardiac repair. Despite promising preclinical results, challenges such as low cell retention, immune rejection, and inconsistent clinical outcomes persist. Recent advancements in genetic engineering, and innovative delivery methods, including transendocardial and intracoronary injections, offer new avenues for enhancing therapeutic efficacy. This review underscores the need for further research to optimize stem cell-based therapies, improve clinical trial design, and translate these innovative approaches into effective treatments for heart disease. By addressing these challenges, stem cell therapy holds the potential to revolutionize cardiac regeneration and improve outcomes for patients with ischemic heart disease and heart failure.

Emberger syndrome (ES), an autosomal dominant disorder characterized by congenital deafness, primary lymphedema, and predisposition to myeloid malignancies, is caused by mutations in the GATA2 gene. Although primary lymphedema is an important hallmark of ES, the pathophysiology remains unclear due to the lack of a suitable experimental model. In this study, we isolated induced pluripotent stem cells (iPSCs) from two patients with ES (i.e., ES-iPSCs) and analyzed their in vitro lymphatic differentiation potential via the mesodermal progenitor stage. KDR+ CD34+ early mesodermal progenitors generated from either ES-iPSCs or wild-type iPSCs during a 6-days serum- and feeder-free culture supplemented with bone morphogenetic protein 4 and vascular endothelial growth factor (VEGF) had almost equivalent developmental potential. However, upon co-culture with OP9 stromal cells, KDR+ CD34+ cells derived from ES-iPSCs developed into CD31+ lymphatic vessel endothelial hyaluronan receptor-1+ VEGF receptor 3+ lymphatic endothelial cells less efficiently than KDR+ CD34+ cells derived from wild-type iPSCs. Thus, patient-derived iPSCs recapitulate impairments at an early stage of lymphangiogenesis, making them a useful experimental tool for dissecting the pathophysiology of primary lymphedema in ES and developing potential therapeutic approaches.

Inflammatory bowel disease (IBD) is a chronic inflammatory disorder of the gastrointestinal tract caused by dysfunction of the immune system in genetically susceptible individuals. As current pharmacologic and surgical treatments remain suboptimal, increasing attention has been directed toward exosomes derived from mesenchymal stem/stromal cells (MSCs) as alternative therapeutic approaches. MSCs are multipotent stromal cells that can be isolated from various human tissues such as bone marrow, adipose, umbilical cord and periodontal ligament. Exosomes are cell-derived membrane-bound vesicles enclosing RNAs, proteins, growth factors, and cytokines. Previous studies indicate that the anti-inflammatory, immunomodulatory, and regenerative effects of MSCs are largely mediated by MSC-derived exosomes (MSC-Exos). Therefore, this review outlines current insights into the molecular mechanisms of MSC-Exos in IBD treatment to support the future development of MSC-Exos as a therapeutic strategy, thus providing novel observations into the clinical applications of MSC-Exos in IBD management.

The leading factor contributing to patient mortality is the local invasion and metastasis of tumors, which are influenced by the malignant progression of tumor cells. The epithelial-mesenchymal transition (EMT) is key to understanding malignancy development. EMT is a critical regulatory mechanism for differentiating cell populations initially observed during the neural crest and embryonic gastrulation formation. This process is closely associated with tumor metastasis in cancer and is also related to the maintenance of cancer stem cells. Flavonoids, known for their antioxidant properties, have been widely studied for their anticancer potential to protect plants from harmful environmental conditions. They have attracted considerable attention and have been the focus of numerous experimental and epidemiological studies to evaluate their potential in cancer treatment. In vitro and in vivo research has demonstrated that flavonoids can significantly impact cancer-related EMT. They may inhibit the EMT process by reducing the levels of Twist1, N-cadherin, ZEB1, integrins, SNAI1/2, CD44, MMPs, and vimentin while increasing E-cadherin levels and targeting the PI3K/AKT, NF-κB p65, and JAK2/STAT3 signaling pathways. In order to suppress the transcription of the E-cadherin promoter, several Zn-finger transcription factors, such as SNAI2, ZEB1, and ZEB2, and basic helix-loop-helix (bHLH) factors, such as Twist, may directly bind to its E-boxes. Overall, clinical cancer research should integrate the anticancer properties of flavonoids, which address all phases of carcinogenesis, including EMT, to improve the prospects for targeted cancer therapies in patients suffering from aggressive forms of tumors.

Neural-activated optogenetics technique contributing to the "restart" of degenerative neurons offers hope for the treatment of several neurodegenerative diseases. However, the limitations of persistent invasiveness and inadequate repair of the pathological environment strongly hinder its further application. Here, a concept of differentiating stem cells is proposed to produce functional materials to enhance the therapeutic applicability of optogenetics. Induced pluripotent stem cells (iPSCs) are differentiated to generate the "tentacled" stem cells TenSCs. Their "tentacled" vesicles TenSCev, upon inheriting the biological functions of the parent cell, will possess both neural targeting capacity and pathological environment repair ability. Hence, TenSCev are utilized as functional carrier to deliver optogenetics elements for completely non-traumatic and controllable neuron activation, while also facilitating the comprehensive restoration of the pathological environment, thus effectively halted disease progression and significantly improved cognitive function in Alzheimer's disease or aged mice. Further, the concept of generating specialized biomaterials from differentiated stem cells as functional carriers holds the potential to broaden the applicability of various neuroregulatory techniques in the treatment of neurological disorders.

Cell cycle structures vary significantly across cell types, which exhibit distinct phase compositions. Asynchronous DNA replication and dynamic cellular characteristics during the cell cycle result in considerable heterogeneity in DNA dosage, chromatin accessibility, methylation, and expression. Nonetheless, the consequences of cell cycle disruption in the interpretation of multi-omics data remain unclear. Here, we systematically assessed the influence of distinct cell phase structures on the interpretation of omics features in proliferating cells, and proposed solutions for each omics dataset. For copy number variation (CNV) calling, asynchronous replication timing (RT) interference induces false CNVs in cells with high S-phase ratio (SPR), which are significantly decreased following replication timing domain (RTD) correction. Similar noise is observed in the chromatin accessibility data. Moreover, for DNA methylation and transcriptomic analyses, cell cycle-sorted data outperformed direct comparison in elucidating the biological features of compared cells. Additionally, we established an integrated pipeline to identify differentially expressed genes (DEGs) after cell cycle phasing. Consequently, our study demonstrated extensive cell-cycle heterogeneity, warranting consideration in future studies involving cells with diverse cell-cycle structures. RTD correction or phase-specific comparison could reduce the influence of cell cycle composition on the analysis of the differences observed between stem and differentiated cells.