The chemical investigation of a CH2Cl2:MeOH (1:1) extract of the stem bark of Erythrina caffra resulted in the isolation and characterisation of the novel cerebroside, erythroside (1), alongside eight other known compounds (2-9). The isolated compounds were characterized by FT-IR, 1D and 2D NMR spectroscopy, and ESI-MS analysis. Of the eight known compounds, 2, 3, 5, and 7 are isolated for the first time from the genus Erythrina. The crude extract, its fractions, and selected isolated compounds were evaluated for cytotoxicity against three normal cell lines, human keratinocytes (HaCaT), human melanocytes (NHEM-Ad), and Human Embryonic Kidney 293 (HEK293), using the resazurin/Alamar blue and crystal violet assays. Antioxidant potential was also assessed through both the oxygen radical absorbance capacity (ORAC) and the Trolox equivalent antioxidant capacity (TEAC) assays. Most compounds showed no cytotoxic effects on these cell lines. The antioxidant assay revealed that compound 1 had a moderate antioxidant effect in the ABTS assay (0.50 µmol TE/mg) and good antioxidant capacity in the ORAC assay (0.87 µmol TE/mg). In contrast, almost all these compounds and extracts have shown moderate to good antioxidant effects in the ORAC assay. The results offer valuable insights into the chemical constituents of Erythrina caffra stem bark, their effects on HaCaT, NHEM-Ad, and HEK 293 cells, as well as their antioxidant potential. Future research could focus on identifying additional bioactive compounds within the ethyl acetate fraction that may be responsible for the observed toxicity.

Mesenchymal stem cells (MSCs) are multipotent progenitor cells with the ability to differentiate into several cell types that hold great promise for therapeutic applications. However, the maintenance of proliferative and stemness capacity following in vitro expansion remains a significant challenge. Triiodothyronine (T3) plays a crucial role in embryogenesis and fetal development, yet the knowledge of its effects on MSCs' survival and function is limited. Here, we investigate the impact of T3 treatment in bone marrow (BM)-MSCs and adipose tissue (AT)-MSCs isolated from C57BL/6J mice, to assess stemness preservation, proliferation, and gene expression during in vitro expansion. To this end, MSCs were treated with T3 at various concentrations for 24 and 48 h, and thyroid hormone-responsive and stemness-related genes expression, proliferation, clonogenic potential, and surface marker profiles were analyzed using reverse transcript quantitative PCR, Cell Counting Kit-8, colony-forming unit-fibroblast assays, and flow cytometry. Our results show that T3 exposure did not affect variability or clonogenic potential of BM-MSCs and AT-MSCs, and the expression of T3-responsive genes is activated by distinct time- and dose-dependent responses to T3 in AT-MSCs and BM-MSCs. However, in BM-MSCs, a transient increase in pluripotent markers was observed. Conversely, AT-MSCs exhibited sustained increases in Nano-g, Sca-1, and Ssea-1, particularly at 10 and 100 nM. Collectively, we observed that T3 exposure during in vitro expansion enhanced stemness features in MSCs. This finding was more prominent in AT-MSCs compared with BM-MSCs. The data suggest that T3 exposure during AT-MSCs expansion could be a valuable tool to increase the yield and stemness of these MSCs, facilitating their therapeutic use.

Growth traits in pigs are governed by complex polygenic architectures, with most associated loci residing in non-coding regions that exert substantial influence on economically relevant phenotypes. However, the molecular mechanisms underlying these regulatory elements remain poorly characterized. In this study, a non-coding mutation-designated as NR2C2 recognition motif sequence variation (NRMSV), located 2083 bp upstream of the HMGA1 gene-was identified as a functional modulator of growth traits in a three-generation Eurasian hybrid pig population. NR2C2 is a nuclear receptor implicated in skeletal development and metabolic regulation, while HMGA1 is a key determinant of body size across mammalian species. In embryonic fibroblasts, where NR2C2 is abundantly expressed, the mutant NRMSV suppressed transcriptional activity, functioning as a silencer. In contrast, in bone marrow mesenchymal stem cells, characterized by low NR2C2 expression, the same allele acted as a robust transcriptional enhancer. Knockdown of NR2C2 in embryonic fibroblasts abrogated this repression and restored enhancer activity, confirming the context-dependent, bidirectional regulatory effect of NRMSV on HMGA1 expression. These findings establish the NR2C2-NRMSV-HMGA1 pathway as a novel regulatory mechanism underpinning phenotypic variation in pig growth traits, offering mechanistic insights into mammalian developmental regulation and informing targeted genomic selection for improved productivity in porcine breeding programs.

Glioblastomas (GBMs) are highly aggressive and therapy-resistant brain tumors, mainly driven by stem-like cells and profound metabolic plasticity. Novel treatment strategies, including mechanical high-intensity focused ultrasound (mFUS), are being developed, but their effects on tumor metabolism remain poorly understood. To address this gap, we investigated the impact of mFUS on carbohydrate metabolism in patient-derived GBM organoids and 3D glioma stem-like cell (GSC) cultures. We showed that mFUS selectively induced the expression of glycolysis- and metabolite-transport-associated molecules (GLUT1, HK2, PKM2, LDHA, MCT1, MCT4), particularly in GSCs, as confirmed by qPCR and immunofluorescence. Functional assays demonstrated increased glucose uptake after mFUS, while lactate production remained unchanged. Notably, pharmacological inhibition of GLUT1 or MCT1 potentiated the cytotoxic effects of mFUS, significantly reducing the survival of peri-focal GSCs. Together, these results reveal that mFUS promotes metabolic adaptations in GBM cells and that combined metabolic inhibition may enhance its therapeutic efficacy.

The intestinal mucosa is a complex functional layer which is formed from a diverse range of cell types that include epithelial cells (within crypts and villi) and an array of mesenchymal cells. Many intestinal diseases involve loss of the surface mucosa which can be difficult to restore, and which delays healing and return to normal function. We reason that development of a transplantable intestinal mucosal tissue graft may be a potential therapeutic strategy to aid healing. To be clinically useful, such a tissue graft would need to be capable of rapid production, avoid the risk of host rejection and be demonstrably safe. To create a potential intestinal graft, we developed a novel early-stage human induced pluripotent stem cell (hiPSC) co-differentiation platform capable of generating multiple intestinal cell lineages (epithelial, mesenchymal and endothelial) in 8 days. This protocol is simple to implement, serum-free and greatly reduces the use of animal products. We confirmed the identity of cells by demonstrating that these cells had RNA and protein expression profiles typical of intestinal cell lineages. In particular, we used bulk and single-cell RNA sequencing to characterise global cellular transcriptional profiles robustly and showed that the cells have intestinal identity with early polarisation towards colonic differentiation. The results were replicated across multiple hiPSC lines and in an independent centre. We further cultured the derived cells on collagen hydrogels to form colon-like intestinal patches (CL-IPs). When transplanted into mouse subcutis, CL-IPs formed into colon-like tissue structures, including crypts, stromal and muscle layers. They also developed human-origin vasculature which underwent anastomosis with the murine vasculature to transport murine blood into the graft. Teratoma assays and molecular analyses showed no evidence of residual pluripotency. While at an early stage, this platform shows great potential for further development as a potential source for novel intestinal mucosal regeneration therapy. In addition, the platform is physiologically relevant and thus shows promise as the basis for a new generation of in vitro models of intestinal pathobiology.

Acute kidney injury (AKI) remains a major clinical challenge due to the lack of effective interventions. While mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) show therapeutic promise for AKI, their exact mechanisms are largely to be understood.

The therapeutic efficacy of osteoporosis (OP) treatments is often limited by inadequate cellular precision and poor accumulation within the bone microenvironment. Although synthetic nanoparticles have been developed to address these challenges, they commonly face biological barriers such as rapid systemic clearance, inefficient transendothelial transport, and limited affinity for the complex bone niche. Here, we report on a biomimetic nanobiotechnology platform that integrates biological recognition with precision polymer engineering to overcome these limitations. We engineered a core-shell nanostructure consisting of a bilirubin-loaded Poly D L-Lactide-co-glycolide (PLGA) core cloaked with genetically modified osteoblast (OB)-derived membranes overexpressing the Ephrin type-B receptor 4 (EphB4) receptor. This biomimetic nanoparticle (NP) exploits the endogenous EphB4-EphrinB2 (EFNB2) signaling axis to achieve selective recognition and preferential uptake by EFNB2-expressing osteoclasts (OCs), displaying significantly higher internalization in OCs compared with mesenchymal stem cells (MSC), macrophages (Mφ), and OBs in vitro. Furthermore, the cell-membrane corona enables efficient transendothelial migration under inflammatory conditions, facilitating targeted delivery to OCs beyond the vascular endothelium. In vitro molecular analyses demonstrated that receptor-mediated NP uptake significantly suppressed key osteoclastogenic regulators, including nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1), cathepsin K, and matrix metalloproteinase-9 (MMP-9). In a preclinical OP model, systemic administration resulted in bone-specific accumulation and robust restoration of trabecular microarchitecture and bone mineral density (BMD). Collectively, this work demonstrates that interfacial nanoengineering can translate complex receptor-guided biological interactions into stable, high-performance nanotherapeutics for the precision treatment of skeletal disorders.

Demineralized dentin matrix (DDM) can serve as a novel scaffold for dental tissue regeneration. This study aimed to assess the impact of two distinct sterilizing methods on morphology of Allo-DDM as well as on the viability, metabolic activity, and alkaline phosphatase (ALP) activity of bone marrow mesenchymal stem cells (BMMSCs).

Tendons are specialized connective tissues with limited intrinsic healing properties due to hypovascularity and low metabolic activity. Mesenchymal stem cells (MSCs) possess regenerative potential for tendon injuries. Among the diverse sources of MSCs, those derived from the bone marrow (BM-MSCs) and adipose tissue (AD-MSCs) are the leading candidates. However, their relative efficacy remains underexplored, particularly in terms of biological characteristics and tenogenic differentiation. In addition, the signaling pathways driving tenogenic differentiation processes remain poorly understood. This study aimed to comprehensively investigate the regenerative potential of AD-MSCs and BM-MSCs for tendon repair.

The Wnt (Wingless/Integrated) signaling pathway is a highly conserved regulator of development, stem cell maintenance, and tissue homeostasis. Its dysregulation is a hallmark of cancer, driving uncontrolled proliferation, epithelial-mesenchymal transition, invasion, and therapy resistance. Increasing evidence shows that Wnt signaling in tumor cells does not operate in isolation but is dynamically shaped by reciprocal interactions with the tumor microenvironment (TME), including fibroblasts, immune and endothelial cells, extracellular matrix, and metabolic stressors. These bidirectional circuits sustain cancer stemness, remodel stromal architecture, and create immunosuppressive and pro-angiogenic conditions that foster tumor growth as well as metastatic dissemination and colonization. In this review, we examine how canonical and non-canonical Wnt pathways intersect with the TME across distinct stages of the metastatic cascade, from local invasion to the establishment of distant niches. We further evaluate therapeutic approaches targeting Wnt signaling and discuss their potential to overcome immune evasion and metastatic progression when combined with immunotherapy or stromal-targeted agents. Finally, we highlight emerging preclinical models, including organoids and tumor-on-a-chip systems, that are advancing our understanding of Wnt-TME crosstalk. Together, these insights position Wnt signaling as a central orchestrator of cancer progression and metastasis and a promising therapeutic target for improving outcomes in advanced cancer.

Host gene expression profiling holds great potential in improving the differential diagnostics of bacterial and viral infections. We investigated its discriminative value in children with suspected serious infections.

The coordination of cell migration and proliferation is essential for embryogenesis and tissue homeostasis. However, the classical gradient signaling model is insufficient to explain how stable mitogenic signaling is maintained within migratory cells. Here, we reveal that primordial germ cells (PGCs) in zebrafish employ migrasomes-vesicular organelles formed during migration-to couple their proliferation with migration, ensuring germline expansion. Migrasomes, generated at retraction fibers via tspan7-dependent biogenesis, deliver the growth factor GDF3 specifically to neighboring PGCs through contact-dependent interactions. GDF3 activates the TGF-β receptor acvr1ba, driving proliferation in a spatiotemporally restricted manner. This homocrine signaling mechanism allows migrating PGCs to autonomously sustain proliferation, circumventing signal dilution in embryonic environments. This work uncovers migrasomes as a bridge linking migration and proliferation, with implications for understanding collective cell behaviors in development and disease.

Mutations in GDP-mannose pyrophosphorylase B (GMPPB) cause dystroglycanopathy, a rare neuromuscular disorder characterized by α-dystroglycan hypoglycosylation, yet the pathogenic mechanisms and therapeutic options remain poorly defined. To dissect the molecular basis of dystroglycanopathy, we generate Gmppb knockout and knock-in (P32L and R287Q) mice. We show that homozygous Gmppb knockout and P32L mutant mice (both male and female) display embryonic lethality, while heterozygous Gmppb-P32L (GmppbP32L/+) mice (both male and female) develop progressive muscular dystrophy accompanied by Purkinje cell loss, peripheral demyelination, and impaired nerve conduction. Integrated biochemical, transcriptomic, metabolomic and glycoproteomic analyses reveal widespread protein hypoglycosylation, metabolic dysregulation and suppressed Wnt/β-catenin signaling, resulting in defective differentiation and regeneration of muscle stem cells. Pharmacological activation of Wnt signaling with CHIR-99021 restores myogenic capacity and improves regeneration after injury. Furthermore, AAV-mediated GMPPB gene replacement reinstates α-dystroglycan glycosylation, normalizes GDP-mannose levels, and rescues motor and electrophysiological defects. Collectively, our findings establish GmppbP32L/+ mice as a faithful model of GMPPB-associated dystroglycanopathy and demonstrate that Wnt pathway activation and AAV-based gene therapy represent promising strategies for treating glycosylation-defective muscular dystrophies.

SLC25A10, the mitochondrial dicarboxylate carrier, plays a crucial role in mitochondrial metabolism and protects against liver lipotoxicity. Moreover, its frequent amplification or mutation in cancers, particularly hepatocellular carcinoma (HCC), correlates with a poor prognosis. This study aimed to investigate the role of SLC25A10 in chemotherapy resistance in HCC and elucidate the underlying mechanisms. In this study, we found that hypoxia increased SLC25A10 expression with a preferential shift toward isoform 3 in HCC. This isoform interacts with the nuclear transporter IPO7 to translocate into the nucleus, where it binds to transcription factor CEBPB. This interaction upregulates the transcription of the anti-apoptotic gene BCL2A1, thereby enhancing HCC cell resistance to the chemotherapeutic agent, etoposide. Notably, disruption of the SLC25A10 isoform 3-IPO7 interaction significantly sensitized HCC tumors to etoposide in vivo, suggesting that targeting this interaction could be a promising therapeutic strategy to improve chemotherapy efficacy in HCC. This study reveals a novel nuclear function of the mitochondrial dicarboxylate carrier SLC25A10 in transcriptional regulation under hypoxic conditions, distinct from its canonical mitochondrial role. These findings expand our understanding of SLC25A10 biology and uncover a previously unrecognized mechanism that drives hypoxia-induced chemoresistance in HCC. Our findings suggest that SLC25A10 is a potential therapeutic target to overcome drug resistance in HCC.

Serum ferritin is an independent prognostic marker in myelodysplastic neoplasms (MDS) and serves as a surrogate parameter for iron overload. Oxidative stress derived from iron overload may induce genomic damage, thereby promoting genetic instability and disease progression in MDS. We aimed to evaluate a possible association between iron overload via serum ferritin and various parameters for genetic instability in MDS.

Expanded potential stem cells (EPSCs) represent a distinct and developmentally primitive stem cell population characterized by their broad developmental potential, which encompasses both embryonic and extra-embryonic lineages. In this study, we employed a polycistronic cassette to directly reprogram human fibroblasts into induced Expanded Potential Stem Cells (iEPSCs). Substituting SOX2 with engineered SOX17 transcription factors resulted in an approximately five-fold increase in the average yield of iEPSC colonies, while maintaining the molecular and functional integrity of the resulting clonal lines. Notably, under feeder-free conditions, SOX2 occasionally failed to reprogram and yielded inconsistent colony numbers, whereas engineered SOX17 and miniaturized SOX17 reproducibly produced feeder-free iEPSCs. In summary, the use of engineered SOX17 enables efficient and robust reprogramming of human fibroblasts into EPSCs, allowing for modeling of early human pre-implantation development, investigating placental disorders, and expanding the toolkit for drug development with a versatile model of pluripotent stem cells that exhibit broader developmental capabilities.

Glioblastoma (GBM) remains a highly aggressive brain tumor with poor prognosis despite surgical resection and standard chemoradiation. Induced neural stem cell (iNSC)-based therapies offer a promising strategy owing to their inherent tumor-homing ability and capacity to deliver therapeutics selectively to the tumor site; however, their poor retention within the tumor resection cavity limits clinical potential. Herein, we evaluated next-generation tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-secreting iNSCs (hiNeuroS) in combination with a biodegradable, thermo-responsive chitosan hydrogel for localized and sustained delivery of high densities of hiNeuroS. Direct comparisons with first-generation iNSCs (hiNSCs) demonstrated that hiNeuroS achieved faster and more extensive tumor cell kill across multiple tumor-to-therapeutic cell ratios in both U87 and patient-derived GBM8 models. Injectable chitosan scaffolds supported stem cell densities up to 2 × 10⁷ cells/mL, maintained > 90% viability, preserved scaffold microstructure, and maintained rapid gelation at physiological conditions. Gravimetric analysis revealed stable 30-day mass change profiles, with minimal net mass loss indicating preserved scaffold integrity. Moreover, encapsulated hiNeuroS retained their migratory capacity and robust TRAIL secretion, inducing significant tumor cell death in both GBM models. Collectively, these findings demonstrate that hiNeuroS maintain functional potency in GBM and are compatible with scaffold-based delivery. This work provides a foundation for future in vivo studies to assess scaffold-mediated retention, persistence, and therapeutic efficacy in an established GBM mouse model, supporting the development of an injectable hydrogel platform for next-generation cell-based therapies for treatment of GBM and other malignant tumors.

Bromodomain (BD) and extra-terminal domain (BET) proteins are key regulators of RNA polymerase II (Pol II)-mediated transcription and their BDs represent promising drug targets. Yet, the interplay between histone acetylation and the chromatin dynamics of individual BET proteins with respect to transcriptional regulation is not fully understood. Here in mouse embryonic stem cells, we uncover an essential role of BRD2 in maintaining Pol II recruitment at promoters through its interaction with TFIID, which becomes particularly critical under the conditions of impaired pause release. Combining rapid protein degradation, chemogenomics and super-resolution microscopy, we show that MOF-mediated histone H4 acetylation promotes BRD2 chromatin association, which in turn enables BRD2 clustering. Accordingly, MOF depletion or deletion of the BRD2's intrinsically disordered region largely recapitulates defects in promoter enrichment and clustering of the transcription machinery observed upon BRD2 loss. Thus, these findings support a model in which histone acetylation-dependent spatiotemporal dynamics of BRD2 coordinate the transcription machinery to regulate transcription initiation.

Emerging evidence supports the maintenance of cellular proteostasis as a key process in resisting age-related diseases. Its intervention by natural compounds is expected to benefit the healthcare system. In the current study, we recruited a unique amyloid beta (Aβ) aggregation-based in vitro screening system and identified fucoxanthin (Fx) as a candidate compound possessing protein de-aggregation potential. Phenotypic and molecular analyses revealed that Fx mitigated endoplasmic reticulum and heat stress by modulating the cellular unfolded protein response and heat shock response, respectively. Furthermore, Fx alleviated senescence-associated markers and restored proteostasis by rescuing chaperone expression and proteasome activity in human senescent fibroblasts. The above protective effect of Fx was further validated in human induced pluripotent stem cells (hiPSC)-derived neurons and Aβ-GFP transgenic Caenorhabditis elegans (C. elegans). Altogether, our results demonstrate the proteostasis-promoting potential of Fx that may be useful for managing degenerative diseases and promoting healthy aging.

Limbal melanocytes (LMs) are neural crest-derived cells that have emerged as multifunctional regulators of ocular surface homeostasis beyond their traditional role in pigmentation and photoprotection. Residing within the basal limbal epithelium, LMs form functional units with limbal epithelial stem/progenitor cells (LEPCs) through cadherin-mediated adhesion and paracrine signaling through soluble factors and extracellular vesicles. LMs contribute to photoprotection by transferring melanosomes to LEPCs, which subsequently organize these organelles into supranuclear melanin caps that may shield LEPC DNA from ultraviolet-induced damage. In parallel, LMs regulate LEPCs stemness, proliferation, and differentiation through direct cell-cell interactions and paracrine signaling mechanisms. LMs exhibit pronounced immunomodulatory properties, including a hypoimmunogenic phenotype, T-cell suppressive capacity, and anti-inflammatory, anti-angiogenic potential. Recent technical advances-such as CD117+CD90- flow cytometric isolation and laminin-based culture systems-have enabled long-term LM expansion. Dysregulation of LMs has been reported in association with several pathological conditions, including corneal inflammation, aniridia-associated keratopathy, pterygium, and rare limbal melanomas. This review integrates current knowledge on LMs embryological origin, anatomical localization, molecular and cellular characteristics, interactions with LEPCs, and functional contributions to ocular surface homeostasis. We discuss the pathological implications of LMs dysfunction and critically evaluate emerging therapeutic strategies, including LMs-enriched tissue-engineered constructs for limbal stem cell deficiency and other ocular surface disorders. By synthesizing the current knowledge and identifying key gaps in understanding, this review aims to guide further research and accelerate the clinical translation of LMs-based regenerative therapies.