CD8+ T cell exhaustion represents a major obstacle to effective cancer immunotherapy. While stem-like progenitor exhausted T (TPEX) cells can differentiate into intermediate (Int-TEX) and terminally exhausted (TEX) subsets, the epigenetic regulation of this process is unclear. We identify the RNA methyltransferase Mettl8 as a critical regulator, with expression significantly higher in TPEX than in TEX subsets. In anti-PD-1 responding non-small cell lung cancer patients, Mettl8 and the stemness factor TCF7 were downregulated. In murine models, Mettl8 deletion restrained tumor progression by driving TPEX differentiation into effective Int-TEX cells. Mechanistically, Mettl8 stabilizes Tcf7 mRNA via m3C modification and enhances Tcf1 protein expression. Additionally, Mettl8 interacts with Tcf1 to facilitate chromatin looping at the Tox locus, maintaining TPEX stemness. Pharmacological Mettl8 inhibition promoted TPEX-to-Int-TEX differentiation and tumor control. Combining this inhibition with anti-PD-1 therapy yielded synergistic efficacy. Our findings establish Mettl8 as a pivotal regulator of TPEX fate and a promising therapeutic target for enhancing immunotherapy.

Psilocybin is studied as innovative medication in anxiety, substance abuse and treatment-resistant depression. Animal studies show that psychedelics promote neuronal plasticity by strengthening synaptic responses and protein synthesis. However, the exact molecular and cellular changes induced by psilocybin in the human brain are not known. Here, we treated human cortical neurons derived from induced pluripotent stem cells with the 5-HT2A receptor agonist psilocin - the psychoactive metabolite of psilocybin. We analyzed how exposure to psilocin affects gene expression, neuronal morphology, synaptic markers and neuronal function. Psilocin provoked a 5-HT2A-R-mediated augmentation of BDNF abundance. Transcriptomic profiling identified gene expression signatures priming neurons to neuroplasticity. On a morphological level, psilocin induced enhanced neuronal complexity and increased expression of synaptic proteins, in particular in the postsynaptic compartment. Consistently, we observed an increased excitability and enhanced synaptic network activity in neurons treated with psilocin. In conclusion, exposure of human neurons to psilocin might induce a state of enhanced neuronal plasticity, which could explain why psilocin is beneficial in the treatment of neuropsychiatric disorders where synaptic dysfunctions are discussed.

Significant progress has been made in research on chronic pancreatitis (CP). It is essential to analyze recent trends in this field to guide future clinical investigations.

Thoracic aortic aneurysms and dissections (TAAD) are life-threatening vascular disorders affecting the medial layer of the aortic wall, associated with high mortality when a rupture or dissection occurs. Though numerous genes are associated with familial TAAD (FTAAD), pathogenic variants in MYH11, encoding smooth muscle cell specific myosin heavy chain (SM-MHC), represent a rare but interesting subgroup as many gaps remain in the knowledge of disease mechanisms, phenotype presentation and gene-environmental interactions. No reliable therapy exists in halting aneurysm growth in affected individuals.

Stem cell biology has rapidly expanded into an interdisciplinary field with potential for next-generation cell-based therapies. However, a critical gap remains in our understanding of how physical forces influence stem cell behavior. Recent studies in mechanotransduction, the process by which cells sense and convert mechanical cues into biochemical signals, have revealed that biomechanical regulation is fundamental for stem cell fate, proliferation, and therapeutic efficacy. This review synthesizes recent findings on the intrinsic and extrinsic parameters of physical cues that govern mechanotransduction and shape stem cell biology, highlights the mechanosensitive responses, and explores how biomedical engineering (BME) can be employed to manipulate these processes to improve translational outcomes in clinical settings. By integrating insights from cell biology, mechanobiology, and engineering, this interdisciplinary field offers strategies to translate benchside discoveries into clinical applications, advancing the development of precise and effective stem cell-based therapies.

After a stroke, many survivors experience post-stroke cognitive impairment (PSCI), a frequent clinical problem that might continue for an extended timeframe. Nerve regeneration is a crucial aspect of the body's self-repair mechanism following a stroke. While Sodium Pyruvate (SP) exhibits notable neuroprotective properties, its potential role in facilitating nerve regeneration requires further investigation.

Human fetal neural stem cells (hfNSCs) from the subventricular zone (SVZ) are employed in clinical trials for neurodegenerative diseases, yet their bioelectrical properties remain largely unexplored. Molecular markers alone do not reliably correlate with functional state, highlighting the need for complementary functional descriptors.

Tumor-associated macrophages (TAMs) represent the most abundant immune cell population within the tumor microenvironment and play a critical role in cancer progression. However, the molecular mediators underlying TAM-driven tumor invasion remain incompletely defined. This study investigated whether ADAM9 functions as a key effector of pro-invasive TAM polarization using a canine mammary tumor model integrated with human transcriptomic datasets.

Dyslipidemia and obesity are key risk factors for cardiometabolic diseases and are also linked to osteoporosis and other bone disorders. Evidence shows lipid metabolism influences bone homeostasis largely through immune regulation. This review first explains how abnormal lipid metabolism disrupts adipogenic and osteogenic differentiation in bone marrow mesenchymal stem cells and alters adipokines like leptin and adiponectin, upsetting bone formation and resorption and leading to bone loss. It then examines the lipid-immune-bone axis. In innate immunity, high lipid levels shift macrophages from M2 to pro-inflammatory M1, increase bone-resorbing cytokines such as TNF-α and IL-1β, and trigger neutrophil senescence and lipid peroxidation with excess reactive oxygen species, all of which promote osteoclast formation and suppress bone growth. In adaptive immunity, hyperlipidemia changes T-cell metabolism, weakens Treg function, and drives Th17 differentiation; this Th17/Treg imbalance boosts osteoclasts via RANKL, IL-17, and related pathways. Meanwhile, in inflammation, B cells switch from producing OPG to releasing RANKL and G-CSF, while Breg-derived IL-10, IL-35, and TGF-β1 protect bone. The review also highlights how M1 macrophages and Th17 cells work together to worsen bone damage. Understanding these immune mechanisms could lead to new treatments for metabolic bone diseases. Despite these advances, the translation of these preclinical findings into clinical practice remains a challenge that warrants further investigation.

Severe combined immunodeficiency (SCID) pigs have become a promising large animal model for biomedical research, offering significant advantages over traditional mouse models due to their anatomical, physiological, and genetic similarities to humans. Humanized SCID pig models can potentially improve preclinical research in areas such as cancer immunotherapies, stem cell therapies, and transplantation methods, yet often lack significant lymphocyte development, including evidence of B cell and myeloid cell development. This work aims to increase the extent of humanization of the SCID pig. CRISPR guide RNAs were successfully developed for the RAG2 and IL2RG genes, and a double-knockout cell line (RAG2-/-IL2RG-/Y, RG) was established. Somatic cell nuclear transfer (SCNT) was then used to create cloned SCID fetuses, which were injected intraperitoneally with in vitro expanded human CD34+ umbilical cord cells at day 41-42 of gestation. Human leukocytes, including T, B, NK, and myeloid cell types, were detected in peripheral blood, spleen, bone marrow and within the thymus of neonatal animals using flow cytometry. Six of the twelve pigs injected had >5% human cells within the CD45+ cell thymic population. Histology of thymus tissues from multiple pigs showed substantial development of the cortex and medulla, which is absent in non-injected RG neonates. This work demonstrates an improvement in the spectrum of xenogenic immune cell lineages developed using an RG line injected with highly expanded CD34+ cells, yet functional analysis of these cell types is needed for further establishment of an in utero humanized SCID pig model.

Colorectal cancer is one of the most prevalent malignant tumors worldwide, and cancer-initiating (CI) cells or cancer stem (CS) cells are critical for tumor progression. We purified CI/CS cells from colon cancer cells utilizing a membrane filtration method. We developed membrane filters with different surface charges (zeta potentials) by blending a negatively ionized polymer [poly(vinyl alcohol-itaconic acid), PVI] or a positively ionized polymer [poly(L-lysine), PLL] into poly(lactide-co-glycolic acid) (PLG). Suspensions of HT-29 colon cancer cells and PAT-3 cells (primary colon cancer cells from a colon cancer patient in this research) were permeated through unmodified PLG filters and modified PLG filters blended with and without PVI or PLL. The cells in the filtration and recovering solutions and migrating cells from the filters after filtration were evaluated to identify the cells in each fraction and which filter enriched CI/CS cells. CI/CS cells were evaluated for (a) CD44 and CD133 (CI/CS cell markers) expression by flow cytometry and immunostaining in vitro. Cells were also (b) evaluated by a colony formation assay in vitro, (c) evaluated for carcinoembryonic antigen (CEA) production by enzyme-linked immunosorbent assay in vitro and (d) evaluated for a xenograft tumorigenicity test using NOD.CB17-Prkdcscid/NcrCrl (NOD-SCID) mice in vivo. The results revealed that the migration of colon cancer cells from positively charged PLG/PLL filters increased the number of CI/CS cells efficiently and killed 83% of NOD-SCID mice after transplantation, whereas the other fraction of cells did not kill NOD-SCID mice. The filtration method through PLG/PLL filters is effective for purifying CI/CS cells from colon cancer cells.

Evans syndrome (ES) is a rare autoimmune cytopenia characterized by autoimmune hemolytic anemia and immune thrombocytopenia. Patients who are refractory to multiple immunosuppressive therapies have limited treatment options, and long-term outcomes remain poor.

Current therapeutic strategies for bone defects, including autografts, allografts, and conventional biomaterial scaffolds, are limited by donor site morbidity, immune rejection, and insufficient vascularization. Moreover, the complex inflammatory microenvironment in bone defects often impairs healing outcomes, necessitating the development of advanced biomaterials with enhanced immunomodulatory and regenerative capabilities. This study investigates the therapeutic potential of extracellular vesicles derived from Astragalus-modified calcium silicate (AstCS)-stimulated M2 macrophages (AstCSM2EVs) in bone regeneration. The AstCSM2EVs demonstrated superior immunomodulatory capabilities by effectively polarizing macrophages toward the M2 phenotype, characterized by significant downregulation of pro-inflammatory cytokines (IL-1β, TNF-α) and concurrent upregulation of anti-inflammatory mediators (IL-4, IL-10). Notably, AstCSM2EVs exhibited enhanced angiogenic potential, evidenced by increased endothelial tube formation and elevated VEGF secretion, while simultaneously promoting osteogenic differentiation of mesenchymal stem cells through upregulated expression of key markers including ALP, BSP, and OC. Mechanistic investigations revealed that AstCSM2EVs modulated these regenerative processes primarily through miR-218-5p-mediated regulation of multiple signaling pathways, including NOD-like receptor and ECM-receptor interaction pathways. In a rabbit femoral defect model, local administration of AstCSM2EVs significantly enhanced bone regeneration, demonstrated by increased bone volume fraction and improved trabecular architecture, while effectively suppressing local inflammation. These findings establish AstCSM2EVs as a promising therapeutic agent for bone regeneration, highlighting their multifaceted roles in immunomodulation, angiogenesis, and osteogenesis. This research introduces an innovative approach that combines extracellular vesicles (EV) with immunomodulatory tissue engineering strategies to improve the treatment of bone defects.

Spatio-temporal regulation of gene expression and cellular behavior is critical for ensuring developmental robustness in all multicellular organisms. The mechanisms regulating robustness are often multilayered, with underlying processes occurring at different spatio-temporal scales. Pluripotent stem cells are maintained in SAMs despite periodic differentiation of stem-cell progeny into all above-ground organs, which together form much of the biomass on Earth. Shoot apical meristems (SAMs) are often exposed to variable growth conditions in nature; therefore, stem cell maintenance must be robust to withstand these changes. We focus our review on the mechanisms that regulate the robustness of the central CLAVATA (CLV)-WUSCHEL (WUS) feedback system, highlighting insights from new studies that integrate biological experiments with mathematical modeling. These studies have revealed the importance of WUS concentration-dependent transcription of CLV3 involving cis-regulatory module and WUS-binding cofactors, CLV3 peptide processing and modifications, multiple signals converging on WUS transcriptional regulation, and WUS protein in regulating its subcellular partitioning, diffusion between adjacent cells, and degradation. We discuss mechanisms that could provide robustness to each of these processes and how their integration could provide tissue-level robustness in stem cell maintenance.

Bone marrow-derived mesenchymal stem cells (BMSCs) are recruited into the gastric cancer (GC) microenvironment and promote progression, though the underlying mechanisms remain unclear. This study investigated how RYK-silenced BMSCs induce GC cell apoptosis, with a focus on the novel role of dihydrocapsaicin (DHC).

As an emerging biological therapeutic approach, stem cell therapy demonstrates broad application prospects in analgesia and tissue regeneration, particularly achieving significant advances in treating conditions such as spinal cord injury and intervertebral disc degeneration. In recent years, preclinical model studies have deepened our understanding of the mechanisms underlying stem cell-mediated pain relief and tissue repair, revealing their potential to regulate inflammatory responses, promote nerve regeneration, and repair damaged tissues through multiple pathways. However, the heterogeneity of preclinical models and the discrepancies between these models and clinical practice, coupled with often insufficient critical appraisal of study quality, remain critical issues requiring urgent resolution in this field. This narrative review systematically summarizes the fundamental theories and key mechanisms underlying stem cell-mediated analgesia and regeneration. It comprehensively evaluates the advantages and limitations of different animal models, critically analyzes major controversies and technical challenges in research, and identifies key directions for future studies. The literature discussed herein was identified through searches in PubMed and Web of Science databases, focusing on recent preclinical studies (primarily within the last decade) involving stem cells, pain models, and tissue regeneration. Selected studies were evaluated for their methodological rigor and contribution to mechanistic understanding. This review aims to synthesize current evidence, critically appraise preclinical models, and provide a forward-looking perspective for research on stem cell-related analgesia and regenerative mechanisms, thereby promoting further development in clinical translation.

Investigations into the anticancer stem cell (CSC) properties of rhodium complexes are extremely rare. Here we report the synthesis, characterization, photophysical properties, and in vitro anti-CSC activity of a series of cyclometalated rhodium-(III)-polypyridyl complexes 1-4. The 4,7-diphenyl-1,10-phenanthroline-bearing complex 4 exhibits activity against monolayer- and three-dimensionally cultured breast CSCs and osteosarcoma stem cells in the sub-micromolar to low micromolar range, outperforming the metallodrug cisplatin and the established anti-CSC agent salinomycin. Our results suggest that the reported rhodium-(III) scaffold, containing cyclometalated 2,2'-(phenylmethylene)-dipyridine, could be a useful synthon for the future development of anti-CSC rhodium complexes.

At the Wisconsin National Primate Research Center, we have identified a family of rhesus carrying the microtubule-associated protein tau ( MAPT ) R406W mutation linked to frontotemporal dementia (FTD). Rhesus induced pluripotent stem cells (RhiPSCs) derived from these monkeys present a unique opportunity for in vitro modeling and comparison with cells derived from MAPT R406W human carriers. Here, we report the development of a reproducible method to generate RhiPSCs compliant with the standards of the International Society for Stem Cell Research (ISSCR) to support in vitro modeling of FTD -MAPT R406W. Our stepwise approach identified efficient methods for fibroblast derivation, fibroblast reprogramming to RhiPSC, and RhiPSC maintenance over continued culture. To derive fibroblasts from MAPT wild type (WT) and R406W monkeys, a combination of manual processing and overnight enzymatic digestion was required to maximize the number of low passage fibroblasts available for reprogramming. Fibroblast reprogramming to RhiPSC using Sendai viral vectors versus oriP/EBNA1 episomal plasmids revealed the latter as most efficient. Electroporation conditions for oriP/EBNA1 reprogramming were optimized to maximize plasmid uptake and cell survival. Ultimately, eight RhiPSC lines were derived from 4 donor rhesus monkeys (n=2 WT, n=2 R406W; two clonal lines per donor) and fully characterized according to ISSCR standards. RhiPSC stemness and genetic stability was best maintained on mouse embryonic fibroblast feeders in Universal Primate Pluripotency Stem Cell medium, as opposed to Essential 12 medium supplemented with IWR1, which produced cytogenetic abnormalities. Rhesus neural progenitor cells were generated using a monolayer protocol and expressed PAX6 and NESTIN after 21 days of differentiation. Our reliable method will be useful to labs seeking to derive RhiPSCs for preclinical studies. Overall, the RhiPSCs generated from MAPT R406W carriers will be a critical resource for evaluating the molecular underpinnings of tau-related neurodegeneration across primate species.

The activation of cis-regulatory enhancers is essential for cell fate specification by driving cell type-specific gene expression. Differentiation models are widely used to study enhancer biology but the asynchronous and interdependent nature of gene regulatory changes during cell state transitions can complicate mechanistic studies. To overcome these limitations, here we develop a tamoxifen-gated system for acute enhancer activation in embryonic stem cells (ESCs) based on GRHL2, a pioneer transcription factor which naturally becomes expressed as naive ESCs differentiate into the formative ESC state. Using this system, we investigate the functional relationship between GRHL2 and the histone mono-methyltransferases KMT2C and KMT2D (KMT2C/D). GRHL2 readily binds its target sites independent of KMT2C/D. However, in the absence of KMT2C/D, there are dramatic reductions in H3K4me1/2, P300 recruitment, and H3K27ac deposition at these sites as well as diminished transcriptional activation. Still, strikingly, a basal level of active enhancer mark acquisition and transcriptional activation occurs. Consistent with these findings, during the naive to formative ESC differentiation, GRHL2 enhancer remodeling and target expression is also strongly but incompletely dependent on KMT2C/D. Together, these results define a functional co-activator relationship in which KMT2C/D act as important amplifiers of GRHL2-driven enhancer activation in ESCs and establish a rapid inducible system for dissecting the kinetics and enzymatic dependencies of pioneer transcription factor mediated enhancer remodeling.