The development of tissue-engineered heart valves (TEHVs) remains a challenge worldwide. In this study, a series of double-crosslinked methacrylated (MA) carrageenan (CA)/MA gelatin hydrogels loaded with heparin (CA/Gel@Hep) were developed as potential materials for TEHV. CA/Gel@Hep hydrogels with different concentrations of heparin were fabricated, and their mechanical properties, swelling/degradation behaviors, and heparin release profiles, and biological performance were systematically studied. The rheological tests showed that storage modulus (G') was consistently higher than loss modulus (G''). Unconfined compression tests showed that the compressive modulus of CA/Gel@Hep hydrogels ranged from 0.26 ± 0.01 to 0.43 ± 0.03 kPa, which matches the mechanical requirements of native valve leaflets.In vitroevaluation demonstrated that CA/Gel@Hep hydrogels exhibited good cytocompatibility and blood compatibility, excellent anticoagulant properties, and facilitated migration and proliferation of human adipose tissue-derived mesenchymal stromal cells. quantitative PCR results showed that CA/Gel@Hep hydrogels significantly upregulated the expression of genes related to valve remodeling, including SMA, VIM, MMP1, and MMP2. These results suggest that CA/Gel@Hep hydrogels hold great potential for heart valve tissue engineering applications.

Vascular cambium produces cells to form xylem, or wood, in tree stems. Here, we explored transregulatory pathways in this cell lineage development. We tested 20 of the 95 putative vascular cambium-specific (VCS) transcription factor genes through gain-of-function transgenesis in Black cottonwood (Populus trichocarpa) and found that a SHORT-ROOT (PtrSHR1) gene induced some of the most prominent phenotypes. PtrSHR1 transcripts are in the vascular cambium (VC) but not in the stem-differentiating xylem (SDX), whereas PtrSHR1 proteins are in both VC and SDX. Constitutive overexpression or endonuclease-deficient Cas9 (dCas9)-mediated loci-specific gene activation of PtrSHR1 revealed an activated PtrSHR1-PtrWRKY25-PtrVCS2-PtrWOX4a/b (WUSCHEL-related homeobox 4a/b) regulatory path for an abbreviated VC zone and a smaller stem diameter. CRISPR PtrSHR1 mutation reduced the PtrSHR1-PtrWRKY25-PtrVCS2-PtrWOX4a/b regulation resulting in an expanded VC zone and a larger stem diameter. In SDX, PtrSHR1-PtrWRKY94 activated specifically 3 (Ptr4CL3, 4-coumarate:CoA ligase 3; PtrC3H3, p-coumarate 3-hydroxylase 3; and PtrC4H1, cinnamate 4-hydroxylase 1) of the 22 monolignol biosynthetic pathway genes to control lignin content and structure, corroborated by chemical degradation and 2-dimensional (2D) nuclear magnetic resonance analyses. Thus, PtrSHR1 mediates the proliferation of VC and diffuses into SDX to control lignin properties and wood production, unveiling potential strategies for creating advantageous wood feedstock for materials and energy.

In patients with low-grade gliomas (LGGs), reallocations of cortical functions (i.e., plasticity) evolve over the course of the disease, allowing serial resections while preserving patients' neurological status. This study aimed to capture evolving patterns of LGG-induced plasticity by means of longitudinal measures of cortical functions based on serial intrasurgical direct electrical stimulation (DES) mappings and navigated transcranial magnetic stimulation (nTMS) mapping. It further assessed nTMS prediction accuracy using DES measures as a reference.

Hexafluoropropylene oxide dimer acid (HFPO-DA), commercially known as GenX, was introduced as a potentially safer substitute for an older type of per- and polyfluorinated substance (PFAS) named perfluorooctanoic acid (PFOA). Emerging evidence suggests that GenX may possess neurotoxicity comparable to or greater than that of PFOA, underscoring the need for evaluating its potential to induce adverse health effects on the central nervous system. Here, we performed a systematic evaluation of predifferentiation GenX exposure and its neurotoxic effects utilizing human induced pluripotent stem cell (hiPSC)-derived cortical neurons. Neurons exposed to 0.4 and 4 ppb GenX prior to differentiation possess altered neuronal characteristics including synaptic density and neural activity, accompanied by transcriptomic changes associated with neurodegeneration, including enriched differentially expressed genes (DEGs) in the Alzheimer's disease (AD) pathway and predicted dysregulation of amyloid processing. Consistent with the transcriptomic alterations, GenX exposure altered multiple APP processing readouts, including increased sAPPβ/sAPPα ratios and intracellular C99 accumulation, accompanied by reduced extracellular Aβ40 and Aβ42 levels. Hyperphosphorylation of tau was also observed along with lipid droplet accumulation and reduced global translational activity, indicating broader disruptions. Collectively, our findings suggest that GenX exposure prior to differentiation, mimicking developmental exposure, can lead to persistent molecular and functional alterations in human cortical neurons that resemble key features observed in neurodegenerative diseases.

Alveolar echinococcosis (AE) is a lethal zoonosis caused by infiltrative growth of the metacestode larva of the tapeworm Echinococcus multilocularis in host organs. We previously showed that the Echinococcus metacestode is an evolutionarily unique, broadly posteriorized tissue, leading us to hypothesize that canonical WNT (cWNT) signalling, which patterns the body axis across metazoans, might be critical for metacestode formation. Here, we report effective RNAi-mediated knockdown of the E. multilocularis β-catenin gene (bcat-1), the central effector of cWNT signalling, in a primary parasite cell culture system that produces metacestode vesicles. bcat-1(RNAi) cultures were markedly impaired in vesicle formation, exhibited stem-cell hyperproliferation, and displayed changes in muscle-fibre organisation. Genome-wide transcriptomics revealed a general anteriorization of gene expression, and in situ hybridization showed an overproduction of cells expressing head-inducing factors such as sfrp upon bcat-1 knockdown. Conversely, metacestode-specific genes including the tegumental factors muc-1, TNFR, and antigen B as well as the posterior marker post2b were significantly downregulated, consistent with the observed vesicle-formation defects. In situ analyses further identified anterior markers frizzled-10, nou-darake, notum, and follistatin that were overexpressed in bcat-1(RNAi) cultures and localized to the future anterior pole at the earliest stages of protoscolex formation. Together, these findings establish a central role for cWNT signalling in directing Echinococcus body-axis formation and the posteriorization events driving metacestode growth within the host, providing insight into asexual parasite proliferation mediated by this biologically unique larval stage and pointing to potential targets for chemotherapy against AE.

Epstein-Barr virus (EBV) is associated with multiple malignancies including Burkitt lymphoma (BL), Hodgkin's lymphomas, nasopharyngeal carcinomas (NPC), and gastric cancers. Canonically, EBV positive tumors display latent gene expression programs that are difficult to target pharmacologically. To overcome this hurdle, lytic reactivation therapies have been developed based on HDAC inhibition with limited mechanistic studies. We therefore characterized the impact of pan-HDAC inhibitor, panobinostat, and class I HDAC inhibitor, nanatinostat, on the growth, survival, and lytic reactivation of four EBV-positive cell lines: P3HR1-ZHT BL, Jijoye BL, IBL-1 immunoblastic lymphoma, and de novo infection derived lymphoblastoid cell lines (LCL). All lines were sensitive, enabling us to define ranges of sensitivity within which to use single cell approaches to assess early EBV lytic gene expression, cell cycle state, and apoptosis. We observed that each EBV-positive model of malignancy responded uniquely to the same HDAC inhibitors and that lytic reactivation was successful in only a small percentage of the cell population. To elucidate the potential role of host factors in preventing successful lytic reactivation, we performed single-cell RNA sequencing on the P3HR1-ZHT BL line treated with the HDAC inhibitor panobinostat. We observed that abortive lytic cells, or cells that do not successfully progress through the lytic cycle, upregulated genes downstream of NF-κB activity. Additionally, genes involved in immune signaling including the CD137/CD137L signaling axis, were upregulated in abortive lytic cells. Functional validation through a Cas9-RNP approach revealed that the CD137 receptor is indeed involved in preventing successful lytic reactivation. These data have important implications for how we approach oncolytic therapies for EBV-associated malignancies.

The immunomodulatory properties of exogenous mesenchymal stem cells (MSCs) have been the target of research in immune-mediated diseases and organ transplants. However, the altered microenvironment decrease MSCs capabilities and survival post-transplantation. This study investigated the viability, proliferation, gene expression and proteomic of canine adipose tissue-derived MSCs (cAT-MSCs) treated with deferroxyamine [DFO] (hypoxia), interferon-γ [IFN-γ] (inflammation) or both for 48h. At 24 hours, all groups exhibited fibroblastoid morphology and adhesion to plastic, with treated groups showing greater cell spacing. After 144h, cell proliferation did not differ significantly between groups, though the treated groups had higher cell concentrations compared to the control. Gene expression analysis revealed increased Casp9 expression in the IFN-γ group, in comparison to the IFN-γ + DFO group; the FGF2 gene was upregulated in the IFN-γ group, while the DKC1 and PT53 genes showed higher expression in IFN-γ than DFO. The VEGFA was more highly expressed in the groups treated with DFO. Proteomics analysis identified 256 proteins, with 70 co-expressed across all groups, and unique proteins in each treatment group: 41 in the control, 44 in DFO, 15 in IFN-γ + DFO group, and 34 for IFN-γ. Notably, 6, 5, and 4 proteins were unique to DFO, IFN-γ + DFO, and IFN-γ treatments, respectively, when compared to the control. Preconditioning modulated angiogenic and metabolic pathways, preserving immunomodulatory function and cellular integrity. Future studies with real hypoxia and multi-omics integration will be crucial for linking molecular signatures to paracrine functions and in vivo efficacy.

Hypoxia during the early post-transplant period represents a major barrier to successful cellular transplantation. This limitation is particularly relevant for pancreatic islet transplantation, a clinical treatment option for diabetes. Stem cell-derived islets are an emerging potential alternative to current primary islets obtained from deceased donors. Although stem cell-derived cells are generally assumed to be more hypoxia tolerant than primary cells, direct quantitative evidence supporting this assumption has been limited, particularly in comparisons between stem cell-derived islets and primary islets. Here, we applied a recently developed pO2_survival metric to objectively compare hypoxia resistance between human primary adult islets and human induced pluripotent stem cell-derived islet spheroids. Using controlled hypoxic culture, live/dead imaging, and computational oxygen modeling, we quantified the pO2_survival as a local oxygen tension at the boundary between viable and non-viable regions within three-dimensional islet constructs. pO2_survival of stem cell-derived islets was significantly lower than that of primary islets (0.01 mmHg vs 2.24 mmHg, P < 0.0001), quantitatively demonstrating enhanced hypoxia resistance of stem cell-derived islet cells. Computational analyses integrating intraspheroidal oxygen distributions and hypoxia resistance further demonstrated improved estimated survival of stem cell-derived islets under large spheroid and hypoxic conditions. Together, these findings provide quantitative evidence that stem cell-derived islets possess enhanced hypoxia resistance compared with primary human islets. This property may expand feasible transplantation sites and reduce early graft loss in stem cell-derived islet therapies.

This study aimed to isolate different cancer cell populations and characterize their secretome profiles to better understand their functional roles in metastasis and tumor progression. For this purpose, we analyzed the secretomes of CD133 and CD326 (EpCAM) positive subpopulations derived from the A549 cell line.

Triple-negative breast cancer (TNBC) exhibits pronounced intratumoral heterogeneity, and cancer stem cells (CSCs) are thought to play a pivotal role in this process. However, the molecular regulatory mechanisms linking CSC-associated stemness features to tumor progression remain insufficiently elucidated.

Progression to castration-resistant prostate cancer (CRPC) is shaped by dynamic interactions within the tumor microenvironment (TME). However, the specific cellular crosstalk driving therapeutic resistance and metastasis remains incompletely defined. This study aims to identify key signaling axes between therapy-resistant luminal progenitor (luminal-2) cells and immune components in the TME, particularly tumor-associated macrophages (TAMs), and to determine how these interactions promote immunosuppression and cancer stem-like cell expansion during disease progression.

Reactive oxygen species (ROS), predominantly derived from mitochondrial respiratory complexes, have emerged as key molecules influencing cell fate decisions like maintenance and differentiation. These redox-dependent events are mainly considered to be cell intrinsic in nature; on the contrary, our observations indicate involvement of these oxygen-derived entities as intercellular communicating agents. In Drosophila male germline, Germline Stem Cells (GSCs) and neighbouring Cyst Stem Cells (CySCs) maintain differential redox thresholds where CySCs have higher redox state compared to the adjacent GSCs. Disruption of the redox equilibrium between the two adjoining stem cell populations by depleting Superoxide Dismutases (SODs), especially Sod1, results in deregulated niche architecture and loss of GSCs, which was mainly attributed to loss of contact-based receptions and uncontrolled CySC proliferation due to ROS-mediated activation of self-renewing signals. Our observations hint towards the crucial role of differential redox states where CySCs containing higher ROS function not only as a source of their own maintenance cues but also serve as non-autonomous redox moderators of GSCs. Our findings underscore the complexity of niche homeostasis and predicate the importance of intercellular redox communication in understanding stem cell microenvironments.

Intestinal ischemia-reperfusion injury (IIRI) is a life-threatening vascular emergency with high mortality, commonly occurring as a complication of hemorrhagic shock or acute mesenteric arterial occlusion. When revascularization is performed after this issue, injury will occur. The influence of MSC-MVs and G-CSF on IIRI was detected in this study. Fifty-six male adult albino rats were allocated into five groups: control, ischemia-reperfusion, MSC-MVs, G-CSF, and recovery. Ischemic injury was induced for one hour using micro-vascular clamp across the superior mesenteric artery at its beginning from the aorta. After that, the clamp was withdrawn to allow reperfusion for two hours. At the end of experimental period, rats were sacrificed. Jejunal tissues were processed for assessment of oxidative stress markers (MDA and SOD) as well as for histological and immune histo-chemical analysis. Ischemia-reperfusion group revealed highly statistically significant increase in tissue MDA while SOD showed the reverse, histologically by light microscopic examination; inflammatory cellular infiltration, congested blood vessels, and separated muscle fibers in muscularis mucosa were observed. Ultrastructurally, absorptive columnar cells appeared with heterochromatic nuclei, lost apical microvilli, and widen intercellular space. These findings showed more improvement in MSC-MVs group in comparison with G-CSF group while recovery group exhibited severe affection.

BEACH-domain-containing proteins (BDCPs) are large scaffolding proteins that regulate vesicle trafficking, autophagy, and granule biogenesis. This review synthesizes recent mechanistic and clinical advances defining BDCP functions in hematopoietic stem and progenitor cell (HSPC) biology, immune regulation, and platelet function, highlighting relevance to human disease.

This study aims to explore the therapeutic efficacy of stem cells from human exfoliated deciduous teeth-derived exosomes (SHED-Exo) in age-related osteoporosis (OP) and clarify its mechanism via mitophagy activation in senescent bone marrow mesenchymal stem cells (BMSCs).

By virtue of their intrinsic immunomodulatory properties, mesenchymal stromal cells (MSCs) represent a promising therapeutic tool for immune-related disorders. Research findings support that MSCs are involved in complex inflammatory pathologies by interacting with local immune cells. In addition to their immunomodulation ability, MSCs also contribute to cell-mediated tissue regeneration due to their potential for multilineage differentiation. However, despite their accessibility, clinical translation of MSCs faces challenges, including their inherent heterogeneity, transient therapeutic effects, and microenvironment-dependent functionality. This review provides an overview of current advances in MSC-based therapies for immune-related disorders, emphasizing Phase III and IV clinical trials and therapies approved by global regulatory agencies. Additionally, we highlight innovative engineering strategies designed to address the limitations of MSCs while enhancing their immunomodulatory capabilities. These approaches include: (1) cell pre-treatment and genetic modification to improve therapeutic efficacy; (2) biomaterial-mediated delivery systems for targeted sites; (3) MSC-derived extracellular vesicle (EV)-based therapeutics to amplify paracrine signaling; (4) induced pluripotent stem cell (iPSC)-derived MSCs to overcome donor variability. By integrating these methodologies with ongoing clinical approaches, this review underscores the potential of engineered MSC immunomodulation in addressing inflammatory pathologies, bridging the gap between basic research and clinical application.

Accumulating evidence emphasizes the importance of microbiota-immune interactions in health and disease development, and identified bacteria-derived small-molecule metabolites as well as macromolecules such as peptides and proteins as promising therapeutic approaches. Here, we identify cytokine motif-containing, immunomodulatory bacterial proteins (CMCPs) as a special category of bacterial proteins in both bacterial genomes and gut metagenomes using Hidden Markov Models (HMMs). We further find eight colorectal cancer‑associated CMCPs differentially enriched in patients or healthy controls. Engineered E. coli Nissle 1917 (EcN) expressing selected CMCPs administered to Apcmin/+ mice selectively colonize intestinal tumors, deliver functional CMCPs in situ, and elicit significant antitumor immune responses while reducing tumor burden. In vitro, purified CMCPs modulate mouse splenic T cells, bone marrow‑derived macrophages and dendritic cells. Our findings indicate that bacterially encoded CMCPs can directly modulate tumor immunity and serve as microbiota‑derived proteins as candidate immunomodulators, which can further be applied in microbiome-mediated immune therapies for CRC.

Thyroid hormone signaling is an essential regulator of skeletal muscle development, function, and metabolism, yet the specific signaling pathways required for muscle regeneration are not yet defined. We used scRNA-seq and the FUCCI (fluorescent ubiquitination-based cell cycle indicator) reporter mouse model to examine how hypothyroidism impacts repair processes after cardiotoxin-induced injury in mice. During regeneration, and up to 2 months after injury, hypothyroid muscles displayed smaller myofibers and a shift to slower oxidative fiber types. scRNA-seq of tibialis anterior muscle during regeneration revealed that hypothyroidism reduced myogenic-lineage diversity. Cell cycle analysis confirmed delayed cell cycle progression at 5 and 14 days after injury, with skeletal muscle stem cells stalled at the G1/S transition, hindering differentiation. Transcriptomic data revealed altered nonmyogenic dynamics, including elevated activated fibro-adipogenic progenitors (FAPs) early in repair and persistent proinflammatory macrophages. Integrative regulon and ligand-receptor analysis further demonstrated that triiodothyronine acted through dual modes: a direct transcriptional control of myogenic cell cycle and oxidative programs and an indirect paracrine remodeling mediated by FAP and immune signaling networks. This study identified what we believe to be novel effects of hypothyroidism on myogenic heterogeneity and impaired tissue repair, offering insights into muscle-wasting mechanisms relevant to hypothyroidism-associated myopathy and sarcopenia.