Search Results (1,332)

Neuroblastomas (NBs) are aggressive, therapy-resistant embryonal tumors of neural crest origin, which despite low mutational burdens exhibit high intra-tumoral heterogeneity characterized by adrenergic, noradrenergic, mesenchymal and cancer stem cell (CSC)-like subpopulations. These phenotypes exhibit interconverting plasticity that reflect both stage of transformation during sympathoadrenal development and conditions within the tumor microenvironment. Chemotherapeutic agents promote adrenergic-to-mesenchymal conversion in NBs, which underpins drug resistance, post-therapeutic relapse, metastatic progression, and the plateauing of responses to advances in multimodal therapy. Improved understanding of the molecular mechanisms that regulate NB phenotypic plasticity is essential for identifying novel prognostic markers and potential therapeutic targets. In this article, following introductions into NB, molecular regulation of NB phenotypic plasticity, and the NF-Y transcription factor and its role in development and differentiation, we focus on alternative NF-YAl, NF-YAs and NF-YAx splicing of the NF-Y subunit, NF-YA, and the potential influence that different NF-YA isoforms have on NF-Y function and the NF-Y-transcription factor networks that impact NB cell phenotypes. Particular attention is paid to the novel extra short-form NF-YAx isoform, originally detected as the exclusive NF-YA isoform in a non-MYCN amplified advanced stage 3 NB. This isoform is also induced by doxorubicin in non-Myc amplified SH-SY5Y NB cells and is involved in doxorubicin cytotoxicity. Despite high cytotoxicity, however, NF-YAx selects a resistant subpopulation with mesenchymal/neural crest stem cell-like identity, unveiling a doxorubicin-induced NF-YAx-dependent resistance mechanism, with potential to influence post-therapeutic relapse and disease progression. Therefore, evaluating alternative NF-YA splicing, and especially NF-YAx expression, in advanced stage and post-therapeutic relapsed NBs, may be of both prognostic and therapeutic significance. Full article

Polycomb repressive complex 2 (PRC2) regulates the expression of pluripotency genes in embryonic stem cells (ESCs) and suppresses multiple genes associated with development, cell fate determination, and differentiation. Mouse embryonic stem cells (mESCs) derived from protein kinase C inhibition (PKCi) exhibit self-renewal and pluripotency comparable to those ESCs captured by the classical 2iL (CHIR99021, PD0325901, and leukemia inhibitory factor) system. However, the dynamic expression pattern of PRC2 in PKCi-mESCs and its role in regulating pluripotency remain unclear. This study demonstrated that the expression level of the enhancer of zeste 2 gene (Ezh2), of which protein is the catalytic subunit of PRC2 responsible for the trimethylation of lysine 27 on nucleosome histone H3 subunit (H3K27me3), is significantly higher in PKCi-mESCs than in 2iL-mESCs. EZH2 knockdown enhances the self-renewal capacity of PKCi-mESCs, as evidenced by a significant increase in the number of undifferentiated mESCs colonies. The effect of an EZH2 reduced expression is accompanied by the upregulation of specific core pluripotency gene Nanog, along with the general downregulation of differentiational genes representing the three germ layers. Conversely, EZH2 overexpression promotes a significant differentiation of PKCi-mESCs, resulting in the downregulation of pluripotency genes, including core pluripotency genes Nanog and Sox2, as well as naïve pluripotency genes Klf4, Fgf4, and Esrrb, while with a wide upregulation of three germ layer associated genes. Importantly, Cleavage Under Targets and Tagmentation (CUT&Tag) demonstrates that EZH2 directly controls H3K27me3 enrichment at the Nanog promoter near the transcription start site. Thus, EZH2, a core subunit of PRC2, exhibits the distinct regulatory functions orchestrating mESCs at a poised state between self-renewal and differentiation under PKC inhibition. EZH2 exerts histone H3 methyltransferase activity to regulate Nanog expression as one of its key targets, thereby modulating the transcriptional regulatory network that maintains pluripotency and lineage specification in mESCs. Full article

Early mammalian embryos are highly sensitive to environmental, metabolic, hormonal, and genomic stress, yet embryo assessment during In Vitro Fertilization (IVF) relies largely on morphology and ploidy for embryo assessment, but these tests incompletely predict miscarriage. We present a transcriptomics based framework to classify and quantify lineage-specific stress in early embryos by benchmarking human preimplantation embryos against dose-, time-, and quality-dependent stress programs defined in Embryonic and placental Trophoblast Stem Cells (ESCs, TSCs) from the implanting blastocyst. Human embryos and stressed ESCs and TSCs are screened using transcriptomic markers from eleven biologically distinct stress Gene Ontology (GO) groups that define functional stress states and enable quantification of pathway presence and upregulation, pathway activity, and downstream outcomes. This framework determines whether the Integrated Stress Response (ISR), once initiated, resolves to enable the Developmentally Associated Stress Response (DASR). High-throughput screening (HTS) titrates stress to define increasingly risky yet biologically equivalent doses for levels of diminished stem cell growth across mechanistically diverse stressors. Then bulk RNA seq derives lineage specific transcriptomic markers putatively respond to common levels of diminished growth and that distinguish weak vs. strong stress and resolved vs. unresolved ISR. These stem cell transcriptomic signatures are applied to bulk RNA seq data from IVF embryos graded for morphology or adhesion, enabling quantitative inference of stress burden, lineage vulnerability, and developmental trajectory. Full article

Skeletal muscle satellite cells (SMSCs) are essential for embryonic myogenesis and postnatal muscle regeneration; however, their cellular heterogeneity and transcriptional dynamics during avian development remain largely unexplored. Here, we performed single-cell RNA sequencing (scRNA-seq) on 42,886 cells isolated from goose leg muscles across four embryonic stages (E13, E15, E18, and E23), with each stage comprising pooled tissues from four female embryos. Unbiased clustering resolved 22 transcriptionally distinct clusters representing six major cell types—satellite cells, myocytes, fibro-adipogenic progenitors, endothelial cells, immune cells, and Schwann cells—with satellite cells being the most abundant. Satellite cells were further subdivided into three functional states (quiescent, activated, and proliferative/differentiating), which followed a continuous, linear pseudotime trajectory from early to late embryonic stages. This trajectory was marked by a progressive downregulation of stemness-associated regulators (e.g., PAX7) and upregulation of myogenic commitment and differentiation factors (e.g., MYF5, MYOD1, and MYOG), faithfully mirroring chronological development. Cell–cell communication analysis revealed that quiescent satellite cells exhibited the most extensive intercellular signaling networks (e.g., FGFR, Ephrin, collagen, CADM), whereas activated and proliferative/differentiating cells showed progressively diminished communication capacity. Across developmental stages, the contribution intensities of key signaling pathways—including SEMA6, CDH, FGF, LAMININ, MK, MPZ, CADM, FN1, and COLLAGEN—varied significantly among satellite cell states, indicating state-specific responsiveness to microenvironmental cues. Collectively, these findings demonstrate that satellite cells dynamically coordinate extrinsic signal integration with intrinsic differentiation programs to achieve orderly myogenic progression. This study provides a high-resolution single-cell atlas of goose SMSC development, uncovering subpopulation heterogeneity, state-specific molecular signatures, and key signaling pathways, with important implications for avian muscle biology and genetic improvement of poultry. Full article

Long non-coding RNAs (lncRNAs) have emerged as important regulators of developmental processes. Recent studies have established roles for lncRNAs in human and murine erythroid regulation, yet additional regulators remain to be discovered. To identify lncRNA candidates involved in human erythroid regulation, we established a pooled genome-editing screen strategy using human embryonic stem cells (hESCs). Long Intergenic Non-Protein Coding RNA 1089 (LINC01089) was selected for functional investigation. We found that reduced LINC01089 expression impaired erythroid differentiation. Transcriptomic profiling revealed consistent downregulation of genes related to hemoglobin assembly, heme biosynthesis, and membrane maturation, suggesting that LINC01089 supports coordinated erythroid transcriptional programs. In particular, progressive reduction of HBB expression emerged as a key transcriptional anchor. Enrichment analyses of upregulated genes identified recurrent focal adhesion signatures, suggesting a potential link between LINC01089 and focal adhesion kinase (FAK)-related signaling. Given prior evidence linking LINC01089 to FAK phosphorylation, we performed a pilot FAK-inhibition experiment, producing a partial shift toward wild-type HBB expression and supporting FAK/phosphorylated FAK (pFAK) signaling as a potential contributing axis in the impaired transcriptional programs. Together, our findings identify LINC01089 as a novel lncRNA linked to coordinated heme–globin transcriptional programs in human erythroid differentiation, with possible involvement of the FAK/pFAK axis. Full article

Maternal consumption of alcohol and drugs during pregnancy can compromise neural development with long-lasting impact on individuals’ health. The inhibitor of growth (ING) family of proteins is an epigenetic regulator that plays a central role in fetal brain development, contributing to neural stem cell maintenance, neuronal differentiation, and the regulation of genes involved in brain morphogenesis. Given the susceptibility of the developing nervous system to epigenetic dysregulation induced by alcohol and drugs, this narrative study aims to summarize literature evidence with the hypothesis that ING proteins may represent a critical but understudied mechanistic link between maternal substance dependence and adverse neurodevelopmental outcomes in newborns. We conducted a comprehensive literature search across three databases (PubMed, Scopus, and Web of Science) up to February 2026 to identify relevant studies. Search terms included combinations of “ING proteins”, “neural development”, “alcohol”, “drugs”, “epigenetic”, “oxidative stress” and “neuroinflammation”. The inclusion criteria were limited to original studies published in English that examined neural development in newborns; the exclusion criteria encompassed non-English publications, letters, editorials, and case reports, and those not directly addressing the specified topics. We identified 55 papers; six were excluded per the exclusion criteria, leaving 49 works discussed in this review. ING proteins are epigenetic regulators essential for embryonic and neural development, including neural stem cell fate and neurogenesis, while substances of abuse are disruptors of the essential pathways necessary for the right fetal brain development. Furthermore, substance abuse creates oxidative stress environments and activates pathways that require ING-mediated chromatin regulation. ING proteins likely act as mediators linking oxidative stress, neuroinflammation, and transcriptional reprogramming in the developing brain. Meanwhile, alcohol and drugs induce epigenetic reprogramming that may disrupt ING-mediated chromatin control. There is little evidence directly linking prenatal exposure (e.g., alcohol and drugs) to ING changes during fetal development. However, we hypothesize that ING proteins function as epigenetic stress response regulators whose disruption by oxidative stress, inflammation, and chromatin alterations induced by prenatal alcohol or drug exposure may contribute to impaired fetal neurodevelopment. Although direct experimental evidence remains limited, this could be a promising and relatively unexplored research area. Full article

Severe mental disorders, including schizophrenia (SCZ), bipolar disorder (BD), and major depressive disorder (MDD), are leading causes of global disability, yet current treatments remain largely symptomatic and fail to alter disease trajectories. Converging evidence from genetics, longitudinal studies, and systems neuroscience supports a dimensional and transdiagnostic architecture of psychopathology, involving shared polygenic risk and overlapping neurodevelopmental and circuit-level alterations. Traditional approaches—such as post-mortem brain analysis, neuroimaging, and animal models—have delineated core molecular perturbations (e.g., dopaminergic, glutamatergic, and GABAergic dysfunction), as well as informed translational frameworks for mechanistic investigation, but remain constrained by restricted access to dynamic processes and incomplete recapitulation of human-specific biology. The advent of human-derived cellular models, particularly human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs), has partially addressed these limitations, enabling the study of patient-specific neurodevelopment and synaptic function in vitro. Within this evolving landscape, the olfactory neuroepithelium (ONE) has emerged as an accessible source of neural progenitors, obtainable through minimally invasive procedures, providing a window into living human neurobiology. ONE-derived cells retain donor-specific genetic and epigenetic signatures while recapitulating disease-relevant phenotypes across major psychiatric disorders, including altered neurodevelopmental dynamics, synaptic gene expression, and inflammatory profiles. Here, we present a narrative review of the principal cellular and tissue models used in biological psychiatry, examining their respective strengths, limitations, and translational relevance across experimental contexts. By situating these approaches within a unified framework, we aim to clarify their complementarity, identify current gaps, and outline future directions, highlighting the emerging potential of ONE-based models to bridge genetic risk, cellular dysfunction, and clinical phenotype, thereby advancing precision psychiatry. Full article

Regulation of all-trans retinoic acid (ATRA) signaling is crucial to early embryonic development. Embryonic stem (ES) cell-derived gastruloids mimic normal development in response to the Wnt/β-catenin agonist CHIR9901, and this study has examined the importance of the activities of RAR (retinoic acid receptor) α and γ to gastruloid development. Expression of retinoic acid receptor (RAR)γ within developing gastruloids was spatially restricted to primitive cells that co-expressed ES cell and early progenitor cell markers, i.e., Nanog, Sox2, and Oct4. In contrast, RARα expression was ubiquitous. mRNAs for the key enzymes involved in ATRA synthesis (Aldh1a2) and degradation (Cyp26a1) were not seen in cells that expressed RARγ. Treatment of ES cell-derived gastruloids with physiologically relevant (10 nm) levels of ATRA or with a highly selective RARγ agonist blocked normal developmental processes, preventing symmetry-breaking and axial elongation. This was not seen following treatments with an RARα agonist, where there was a tendency for enhanced axial elongation. Brachyury (TBXT) immuno-positive cells localized in the posterior end of elongated gastruloids in control- and RARα agonist-treated cultures, with Sox2 immuno-positive cells seen more widely, whilst both TBXT and Sox2 immuno-positive cells were randomly distributed throughout ATRA- and RARγ agonist-treated gastruloids. Concurrent treatment of gastruloids with 10 nm ATRA and 100 nm of an RARγ antagonist partially abrogated the ATRA-mediated block to axial elongation. Conversely, 10 nm RARγ antagonist treatments were associated with the formation of multi-axis gastruloid elongations, with comparatively little effect seen after treatments with an RARα antagonist. These findings reveal that RARγ plays a crucial role in the development of embryonic tissues. Full article

Background: Corneal organoids derived from pluripotent stem cells have emerged as powerful tools for studying corneal development, disease modeling, and regenerative medicine applications. While previous protocols have successfully generated corneal tissue structures, there remains a need for three-dimensional models that recapitulate the complex cellular architecture and diversity of native human cornea. Methods: We developed a modified spontaneous three-dimensional corneal organoid model using human embryonic stem cells (hESCs) through an adapted Self-formed Ectoderm Autonomous Multi-zone (SEAM) protocol. hESCs were cultured as spheroids in ultra-low-binding plates under normoxic conditions and differentiated over 7–8 weeks. Organoids were characterized using immunofluorescence staining for corneal-specific markers and single-cell RNA sequencing to assess cellular composition and gene expression patterns. Results: Approximately 20% of organoids developed transparent regions characteristic of corneal tissue by day 30 of differentiation. Immunofluorescence analysis revealed spatially organized expression of corneal markers, including ZO-1 and E-cadherin in the outermost epithelial layers, P63α-positive putative limbal stem cells at the epithelial–stromal interface, vimentin-positive stromal cells in the interior, and laminin-1 deposition that suggests Bowman’s membrane formation. The organoids expressed cornea-specific keratins (K3, K12, and K15) and the master regulator PAX6 in appropriate cellular compartments. Single-cell RNA sequencing identified 18 distinct cell clusters, including three corneal epithelium subclusters with differential expression of MUC16, KRT12, and ΔNp63α, two stromal populations with distinct inflammatory profiles, and a corneal endothelium cluster. Transcriptomic analysis confirmed expression of key corneal genes, including AQP3, CDH1, multiple keratins, mucins, and extracellular matrix components (HAS2, CD34, CD44, COL8A1, and KERA). Conclusions: This three-dimensional spheroid-based putative corneal organoid model successfully recapitulates the multilayered architecture and cellular diversity of human cornea, including stratified epithelium, putative limbal stem cells, stroma, and endothelium in spatially appropriate arrangements. The model demonstrates molecular signatures consistent with native corneal tissue and provides a valuable platform for studying corneal development, disease mechanisms, and potential therapeutic applications. Future optimization to improve organoid formation efficiency and functional maturation will enhance the utility of this system for both basic research and translational medicine. Full article

Salvia miltiorrhiza Bunge has been used traditionally for cardiovascular disorders, but its specific roles in stem cell cardiac differentiation remain unclear. In this study, we examined whether Salvia miltiorrhiza Bunge (SM) promotes cardiomyocyte differentiation from mouse embryonic stem cells (mESCs) and defined its underlying mechanism. To dynamically monitor cardiac differentiation, we established a Tnnt2-H2B-mCherry reporter mESC line that retained normal pluripotency and differentiation capacity. Using an embryoid body-based differentiation system, we found that SM exerted a distinct temporal effect on lineage progression: treatment during the early differentiation window inhibited pluripotency maintenance, proliferation, and mesodermal development, whereas administration during the cardiac precursor stage markedly enhanced cardiomyocyte formation, as indicated by increased beating embryoid bodies and upregulation of Isl1, Nkx2.5, Tnnt2, Myh6, and Myl7. Mechanistically, transcriptomic and protein analyses showed that SM suppressed canonical Wnt/β-catenin signaling, including downregulation of Dvl2, β-catenin, Axin2, c-Myc, and Cyclin D1, while Wnt activation WAY262611 partially reversed these effects. Further compound screening identified tanshinone IIA (Tan IIA) as the principal active constituent of SM, which largely recapitulated the pro-cardiogenic and Wnt-inhibitory effects of the crude extract. Together, these findings identify SM and Tan IIA as stage-dependent regulators of mESC fate and support their potential utility in natural product-based strategies for improving stem cell-derived cardiomyocyte generation. Full article

Pluripotent stem cells (PSCs) have proven outstanding potential to revolutionize biomedical research. Specifically, their capacity to form 3D multicellular systems that recapitulate organ development and biology, known as organoids, has transformed basic and translational research. The groundbreaking technology is also being applied to the intricate hypothalamus–pituitary (HP) axes, including the target organs (such as gonads, thyroid and adrenal glands). These HP axes govern critical physiological processes, including reproduction, metabolism and stress. Here, we provide an overview of PSC-derived organoid models that are part of the HP axes, both as individual and multi-organ systems, and evaluate their culturing conditions, phenotypic characteristics, advantages, drawbacks and challenges, as well as their potential for disease modeling and therapeutic discovery. By offering this wide perspective, our review will also serve as a key resource for researchers navigating the evolving landscape of PSC-derived organoid technologies in endocrinology. Full article

The potential of water-soluble membrane proteins (wsMPs) has not been fully realized. In this article, we exploit the nearly identical functionality of wsMPs with their membrane-bound counterparts and show that we can create water-soluble membrane proteins that incorporate into the plasma membranes of cells and alter their fate. As a proof of concept, we demonstrate the functional properties of water-soluble engineered pore-forming proteins, K⁺ ionic channels (MthK), and constitutively active GPCRs—among them frizzled receptors—both in vitro and in vivo. We call this method in vivo deployment of recombinant viable MPs, iDRIVE. Furthermore, we demonstrate that our strategy mediates the unidirectional insertion of MPs into the plasma membrane, and through constitutively active receptors, we present evidence for similar signaling pathway activation between small molecules and our water-soluble proteins using model phenotypes and molecular signaling assays. We present three examples where wsMPs are functional in dictating cellular fate, both in vitro and in vivo. Lastly, we show the induction of similar differential methylation via the activation of the Wnt signaling pathway using the conventional small molecule agonist, CHIR99021, or our wsFrizzled receptors (iDRIVE-FZD) in human embryonic kidney (HEK 293) embryoid spheroids (ESs). Additionally, we show that Wnt activation via wsFrizzled receptors results in even more biologically relevant epigenetic changes than via the small molecule CHIR99021. Future work will employ iDRIVE to differentiate stem cells in the production of research and clinically relevant organoids. Full article

Bisphenol A (BPA) has long been used in plastics, resins, and food packaging materials; however, extensive research has demonstrated its reproductive, developmental, and endocrine-disrupting effects. Consequently, BPA has been increasingly restricted and replaced with structural analogues. Among these, tetramethyl bisphenol F (TMBPF) has emerged as one of the most widely used substitutes, particularly in epoxy resins and food-can coatings. Although initially regarded as a safer alternative, accumulating evidence suggests that TMBPF may exert multiple toxicological effects, raising concerns about its potential developmental neurotoxicity. The present study aimed to investigate the neurodevelopmental effects of TMBPF using both in vitro and in vivo approaches. First, a developmental neurotoxicity assay employing Sox1−GFP mouse embryonic stem cells was used to evaluate cytotoxicity using the cell counting kit-8 assay and neural differentiation based on green fluorescent protein (GFP) fluorescence intensity. The results indicated developmental neurotoxic potential according to the established discrimination index. Subsequently, pregnant and lactating mice were exposed to TMBPF daily from gestational day 10.5 to postnatal day 20, and their offspring were assessed for behavioral performance as well as changes in the expression of neurodevelopment-related genes in the brain. Behavioral analyses encompassed multiple domains, including memory and learning, social behavior, anxiety-related responses, and spontaneous locomotor activity, suggesting alterations in these functional outcomes. Molecular analyses further demonstrated changes associated with dopaminergic and cholinergic signaling, synaptic plasticity, neuronal activity markers, neuropeptides, and inflammatory pathways. Collectively, these findings provide the first evidence in a mammalian model that maternal exposure to TMBPF may influence offspring neurodevelopment. These findings suggest potential implications for human exposure to TMBPF, particularly through food-contact materials, and warrant further mechanistic and dose–response studies. Full article

Human cerebral organoids, derived from pluripotent stem cells, are powerful models for studying human brain development. The understanding of how morphogens can be used to guide patterning and differentiation has matured rapidly; however, the influence of basal media components on organoid development remains unclear. Standard organoid media frequently contain non-physiological concentrations of nutrients, including glucose, a central regulator of cellular metabolism and signaling. Here, we examine how glucose availability shapes cerebral organoid growth, morphology, and cell type composition by comparing conventional hyperglycemic media to media with glucose levels more closely resembling normoglycemic conditions. We find that organoids derived from multiple human pluripotent stem cell lines can grow in low glucose, but they exhibit altered growth rates, structural features, and lineage distributions. In H9 embryonic stem cell-derived organoids, inhibition of the mammalian target of rapamycin pathway under low glucose restores neurodevelopmental cell types otherwise diminished in these conditions. These findings highlight glucose as a key determinant of organoid lineage specification and cellular signaling. Importantly, however, glucose modulation does not reduce variability across organoids or cell lines, underscoring the need to better understand and control sources of heterogeneity to improve organoid models. Full article

Background/Objectives: The purpose of this study was to investigate the association between cardiovascular adverse drug events (ADEs) and the use of COVID-19 medicines. Methods: The analyses were conducted by leveraging pharmacovigilance data from the Food and Drug Authority (FDA) Adverse Event Reporting System (FAERS) and TriNetX electronic health records (EHRs). Transcriptomic data from human embryonic stem cell-derived cardiomyocytes (hESC-CMs) exposed to remdesivir were analyzed to provide supportive biological context for the observed cardiovascular safety signals. Results: Comparative analysis of three approved COVID-19 therapies revealed that COVID-19 patients treated with remdesivir had a higher risk of cardiovascular events than those treated with Paxlovid or REGEN-COV. FAERS analysis further indicated that bradycardia, hypotension, and cardiac arrest were the most frequently reported cardiovascular events associated with remdesivir, which was validated by propensity score-matched EHR data. These findings suggest an association between remdesivir exposure and increased cardiovascular ADEs relative to other COVID-19 therapies. Sex-stratified analysis using FAERS and EHR did not show strong sex-dependent patterns for remdesivir-associated cardiovascular ADEs. Age-stratified analyses of EHR data showed age-associated variation across the three cardiovascular ADEs. Bradycardia displayed a non-uniform pattern with higher prevalence in the youngest and oldest age groups, hypotension showed an overall age-associated increase, and cardiac arrest showed only a weak age-associated effect. Pathway enrichment analysis on transcriptomic data revealed that the “cGMP-PKG signaling pathway”, “dilated cardiomyopathy”, and “calcium signaling pathway” were enriched among genes up-regulated by remdesivir exposure. Conclusions: In summary, our integrated analysis of pharmacovigilance, EHR, and transcriptomic data provides convergent evidence for associations between remdesivir and cardiovascular ADEs and offers biological context into these associations. Full article