Paneth cell metaplasia (PCM) is a phenomenon in which Paneth cells, typically found in the small intestine, appear in the colonic epithelium of patients with ulcerative colitis (UC). Our study demonstrates that the PCM occurrence correlates with disease duration and active inflammation. Furthermore, we identified IL-22, an inflammation-associated cytokine, as a key regulator that promotes PCM formation in the colonic epithelium through suppression of Notch signaling and induces REG3A expression within metaplastic niches. In vitro, we show that Reg3a directly enhances cell proliferation and promotes wound healing using mouse colonic organoids. In vivo, Reg3aΔIEC mice in both acute and chronic DSS-induced colitis models exhibit delayed wound healing. Additionally, studies conducted with patient-derived human colonic organoids revealed that REG3A administration stimulates cell proliferation and accelerates wound healing. Together, these findings support a protective role of PCM-associated REG3A in the colonic epithelium of patients with UC.

Disabling hearing impairment is a common human sensory deficit. OTOF is a major deafness gene. It codes for the synaptic protein otoferlin and is essential for transmitter release by inner hair cells (IHCs). Upon genetic loss of otoferlin, cochlear structure and function remain intact up to the IHC synapses, which fail to encode sound. Building on preclinical hearing restoration by AAV-mediated cochlear gene transfer in mice, clinical OTOF-gene-therapy trials are now targeting the pediatric population. However, preclinical optimization and characterization remain urgent needs for the development of OTOF-gene-therapy. Here, we report on the generation and characterization of a marmoset KO that models OTOF-related auditory synaptopathy and can thus address these needs. Following ovary stimulation, harvesting, in vitro maturation and fertilization of oocytes, we injected the zygotes with Cas9 and guide RNAs to disrupt OTOF. Mutant embryos were transferred into the uterus of foster mothers. Marmosets with biallelic, non-mosaic OTOF-KO were normally born and raised by their respective foster parents. Auditory brainstem recordings and otoacoustic emissions revealed profound auditory synaptopathy and OTOF-KO was further validated by the lack of otoferlin expression in IHCs. The new non-human primate model of OTOF-related auditory synaptopathy will serve studies of specificity, efficacy, and longevity of novel inner ear therapies.

Accumulating evidence suggests that neurons are key regulators of skeletal tissues. A recent study by Chen et al. demonstrated that aged brain neurons export WD repeat and FYVE domain containing 1 (WDFY1) to bone via extracellular vesicles. After uptake, WDFY1 engages in retromer-dependent endosome-to-Golgi recycling, shifting bone stem cell fate from osteogenesis toward adipogenesis, revealing a novel mechanism whereby brain aging regulates bone aging.

Genetically modified mesenchymal stem cells (MSCs) have been shown to enhance their therapeutic properties, offering more effective treatment options for various diseases, including metabolic associated fatty liver disease (MASLD). The m7G methyltransferase METTL1 plays a critical role in regulating RNA splicing, stability, and translation. This study presents our findings on METTL1 modified human umbilical cord MSCs, emphasizing their therapeutic effects and the mechanisms involved in treating MASLD.

Camizestrant, a next-generation selective estrogen receptor (ER) degrader and complete ER antagonist, has been associated with a reversible dose- and time-dependent heart rate (HR) reduction in clinical studies. This nonclinical investigation aimed to understand the mechanism of camizestrant-induced HR reduction. The effects of camizestrant on HR in vivo were assessed in rat and dog telemetry studies. Effects on pacemaker channel function in vitro were assessed using patch-clamp electrophysiology in Chinese hamster ovary cells expressing human hyperpolarization-activated cyclic nucleotide-gated channel 4 (hHCN4), human embryonic stem cell (hESC)-derived sinoatrial node (SAN) cardiomyocytes, and primary rat SAN cardiomyocytes. In dogs, 28-day repeat-dose camizestrant administration caused a reversible dose- and time-dependent HR reduction (maximum reduction of 53 beats per min [bpm] on Day 25 vs pre-study levels at 20 mg/kg). HR reduction was also noted in rats (maximum reduction 89 bpm vs vehicle [23%] on Day 5 of a 7-day study at 75 mg/kg). Responses to chronotropic stimuli (e.g., atropine and isoprenaline) were reduced in dogs treated with camizestrant. Camizestrant-induced HR reduction was still present following combined sympathetic (atenolol) and parasympathetic (atropine) inhibition in dogs, as well as vagotomy in rats. Camizestrant reduced hHCN4 current density in Chinese hamster ovary cells, as well as beat rate and "funny" pacemaker (If) current activity in hESC-derived SAN cardiomyocytes. Camizestrant at 75 mg/kg for 7 days significantly reduced If current activity versus vehicle in isolated SAN cardiomyocytes. These results support the hypothesis that camizestrant exerts a pharmacologic, reversible reduction in HR by decreasing SAN pacemaker current activity.

Theranostic nanogels have emerged as a promising class of biomaterials proficient in integrating therapeutic delivery with real-time diagnostic feedback, offering a new level of precision in regenerative medicine. Recent advances in multifunctional nanogel systems have been designed to predict pathological microenvironments, deliver targeted therapeutics, and enable non-invasive monitoring of tissue repair. Key developments in stimuli-responsive drug release, bioinspired surface functionalisation, and advanced imaging combination are critically discussed across applications, including wound healing, cartilage and bone repair, and myocardial regeneration. This review highlights how intelligent nanogels can respond dynamically to biochemical gradients, such as hypoxia, oxidative stress, and inflammatory markers, enabling closed-loop therapeutic control. Despite significant progress, major translational barriers remain, particularly in scalable manufacturing, regulatory harmonisation, long-term biosafety, and reproducibility under GMP conditions. This review concludes that while Theranostic nanogels represent a transformative platform for personalised regenerative therapies, coordinated advances in materials engineering, regulatory science, and clinical validation are essential to realise their full clinical potential.

Depression represents a growing global health challenge, with current pharmacological treatments often exhibiting limited efficacy and significant side effects. Although physical exercise, particularly running exercise, has shown promising antidepressant properties, its underlying neurobiological mechanisms remain poorly understood. In this study, we investigated whether cannabinoid receptor type 1 (CB1R) in the medial prefrontal cortex (mPFC) contributed to the therapeutic benefits of running exercise, which might be associated with astrocyte-related changes. Using a chronic restraint stress (CRS) male rat model of depression, we integrated behavioral assessments (saccharin preference and forced swim tests) with multimodal molecular and cellular analyses, including immunohistochemistry, stereology, immunofluorescence, RT-PCR, and Western blotting. Our findings demonstrate that running exercise alleviated depressive-like behaviors, restored mPFC CB1R expression, and reversed stress-induced reductions in GFAP-positive cells. Conversely, mPFC-specific CB1R knockdown induced depressive-like phenotypes and astrocytic alterations, while CB1R overexpression reproduced some benefits of running exercise. Notably, running exercise failed to improve behavior or astrocytic changes when mPFC CB1R was knocked down, supporting a necessary role for intact mPFC CB1R signaling. These results suggested that mPFC CB1R contributes to running exercise-induced antidepressant effects, and this process is closely associated with the regulation of astrocytic states, providing a potential molecular framework for future therapeutic strategies.

Intestinal epithelial damage and impaired repair are hallmarks of ulcerative colitis (UC), even after inflammation resolves. Intestinal stem cells (ISCs) can retain stable epigenetic changes after inflammation, highlighting the potential for long-lived epithelial memory in the gut. Inflammatory injury in barrier tissues induces epigenetic memory in epithelial stem cells, and the tendency of UC to relapse at previously inflamed sites led us to hypothesize that ISCs from IBD patients acquire lasting memory of prior inflammation. To test this, we derived colonic organoids from inflamed (I) and noninflamed regions of the same UC patients and propagated in long-term culture. Chromatin profiling revealed 2,252 accessible regions unique to I organoids, associated with stress response, repair, and inflammatory genes. Although these regions remained accessible, ∼95% of associated genes were not upregulated in I organoids, indicating a primed state. Upon inflammatory or injury re-challenge, I organoids exhibited heightened transcriptional responses and accelerated wound closure, despite reduced clonogenicity and impaired barrier function, indicating a retained inflammatory memory program. Our findings demonstrate that human ISCs retain a chromatin-based memory of inflammation that persists in the absence of immune cues and shapes future responses to injury. While this may support epithelial adaptation to secondary insults, it may predispose tissue to relapse in patients with UC.

Microelectrode arrays (MEAs) are increasingly used to profile the development of synchronized activity in neural organoids, yet no organoid study has reported on the consistency of electrophysiological development across cell lines. Here, we used dissociated neural organoids derived from six cell lines on MEAs to characterize functional synapse development using multiple parameters across time. The dissociated organoids demonstrated increasing functional connectivity and network activity over time across all cell lines and plasticity in response to synaptic-like stimulation. Like the organoids they were derived from, dissociated organoid cultures contained a diverse mixture of cell types. These results demonstrate that dissociated cerebral organoids can generate functional neurons, akin to primary neuronal cultures from brain tissue, providing a scalable model for studies of neurodevelopment and synaptic function. Consistent with unguided differentiation, we observed variability in activity parameters linked to donor cell line and batch effects, which must be considered in experimental design.

Colorectal cancer (CRC) remains a leading cause of cancer‑related morbidity and mortality worldwide. Although the adenoma-carcinoma sequence and its genetic drivers are well described, the earliest cellular and molecular events initiating tumorigenesis within histologically normal colonic epithelium remain poorly defined. This study aims to identify tumor‑initiating cells (TICs), distinguish them from normal stem‑like cells (nSTMs), and delineate early transcriptional and signaling programs using single‑cell RNA sequencing (scRNA‑seq) from paired normal‑appearing and transformed human colonic tissues.

This research aimed to examine the impact of M2 Macrophages-derived exosomes (M2-exo) on the osteogenic differentiation of Periodontal Ligament Stem Cells (PDLSCs).

The presence of senescent cells in synovial mesenchymal stem cell (MSC) preparations reduces the therapeutic efficacy of MSCs as a treatment for osteoarthritis (OA). Senolytic drugs can selectively kill senescent cells; however, their safety profiles remain a major concern. This study investigated whether cell sorting based on the increased autofluorescence (AF) and cell size of senescent MSCs could selectively eliminate senescent cells from human synovial MSC preparations derived from OA patients.

Persistent inflammatory responses in osteochondral defects produce a hostile microenvironment that severely compromises endogenous tissue repair mechanisms. The local immune microenvironment needs to be modulated to facilitate tissue regeneration. In this study, an innovative bioactive platform was engineered that combined aspirin-induced bone marrow mesenchymal stem cells (BMSCs)-derived extracellular vesicles (Asp-EVs) and a thermoresponsive hydrogel (mPEP) composed of monofunctional polyhedral oligomeric silsesquioxane (mPOSS), polyethylene glycol (PEG), and polypropylene glycol (PPG). We investigated whether mPEP hydrogel-loaded Asp-EVs can enhance osteochondral regeneration by regulating the inflammatory microenvironment and elucidated the underlying mechanisms involved. According to the findings of in vitro experiments, Asp-EVs significantly improved the growth and migration of BMSCs and promoted M1φ-to-M2φ macrophage polarization. Under inflammatory conditions, Asp-EVs restored the chondrogenesis of BMSCs to levels comparable to those of the control group. In vivo, Asp-EVs delivered via the mPEP hydrogel achieved robust osteochondral regeneration in a rat knee defect model, restoring the hyaline-like cartilage and subchondral bone structure. Asp-EVs activated the PINK1/Parkin-mediated mitophagy pathway, rescuing mitochondrial dysfunction and reprogramming the metabolism of macrophages from glycolysis to oxidative phosphorylation and promoting M2φ polarization. These results suggested that the delivery of Asp-EVs via mPEP hydrogels is a promising strategy for osteochondral regeneration.

Stem cell therapies hold promise for halting or reversing kidney disease and improving kidney transplant (KTx) outcomes. One route to large-scale clinical application of stem cell therapies for kidney disease is through their capacity to modulate the balance between tissue injury and repair via crosstalk with other cells. Among the key disease-modulating effects of stem cells is their interaction with components of the immune system involved in harmful inflammation during acute kidney injury (AKI), chronic kidney disease (CKD) and complications of KTx. Extensive basic research demonstrates that stem cells employ diverse paracrine mechanisms to re-program immunological activities from pro-inflammatory/pro-fibrotic to anti-inflammatory/pro-repair. The therapeutic benefits of these effects are confirmed in many pre-clinical models of AKI, CKD and KTx for autologous and allogeneic stem cells including hematopoietic stem cells, mesenchymal stem cells, renal progenitor cells, and induced pluripotent stem cells. Nonetheless, translating these findings into therapeutic immunomodulatory cell products that improve the lives of those with kidney disease is highly challenging. The aims of this review are to: (a) Summarize recent insights into the common molecular and cellular mechanisms of immune-mediated tissue injury in kidney disease and KTx along with the types of stem therapies that have been developed to address them. (b) Critically evaluate the extent to which clinical trials of stem cell products have validated such effects in humans with kidney disease and KTx. (c) Identify key bottlenecks to the large-scale application of stem cell therapies to reduce the burden of kidney disease on patients and societies.

Glioblastoma (GBM), the most prevalent and aggressive primary brain tumor in adults, has a median survival of merely 14 months. Current therapeutic approaches, including maximal safe resection, radiotherapy, and temozolomide-based chemotherapy, have limited efficacy owing to resistance and the high rate of recurrence.

While cell-to-cell variation in gene expression, also known as "transcriptional noise," plays various biological roles, its regulation by genetic variants remains elusive. To address this, we analyzed single-cell RNA sequencing (scRNA-seq) data of induced pluripotent stem cell-derived midbrain cells from 155 individuals together with their genotypes and identified significant quantitative trait loci for transcriptional noise (tnQTLs) genome wide. Among these, tnQTLs without significant effects on expression abundance (termed tn>eQTLs) exhibited characteristics such as drastic alterations under oxidative stress. Analyses using genome-wide association study (GWAS) summary statistics identified enrichment of schizophrenia association signals in tn>eQTLs, as well as putative causal effects of transcriptional noise dysregulation in specific genes. We also analyzed brain scRNA-seq data for schizophrenia and found marked disease-associated transcriptional noise alterations in superficial- and deep-layer excitatory neurons. Overall, this study provides a resource for tnQTLs and insights into the mechanistic basis of transcriptional noise regulation and its relevance to human traits.

Aneuploidy is a hallmark of cancer but often reduces cellular fitness. In childhood B cell acute lymphoblastic leukemia (cB-ALL), hyperdiploidy is the most common cytogenetic abnormality and arises in utero from early hematopoietic stem/progenitor cells (HSPCs), yet its impact on early hematopoiesis remains unclear. We model two proposed routes to hyperdiploidy, chromosome mis-segregation and cytokinesis failure, by transiently exposing human fetal liver-derived HSPCs to reversine or cytochalasin D. Induced hyperdiploidy impaired fitness and delayed differentiation in vitro, causing hyperdiploid cells to be rapidly outcompeted by euploid counterparts. Nonetheless, hyperdiploid cells engrafted immunodeficient mice, where rare clones persisted long term and acquired non-random chromosomal gains frequently observed in cB-ALL. Despite this persistence, they did not initiate leukemia. These findings support a two-step model in which hyperdiploid fetal clones require additional perinatal/postnatal events for malignant transformation. Our work establishes a valuable human model for studying early aneuploidy-driven events in childhood leukemia.

Small intestinal neuroendocrine tumors (SI-NETs) are serotonin-secreting well-differentiated neuroendocrine tumors of enterochromaffin (EC) cell origin. However, EC cell-derived tumorigenesis remains poorly understood. Prior studies using TPH1 Cre-ERT2-driven RPM mice (EC cell targeted RB1 (R) and Trp53 (P) loss and Myc (M) gain) showed non-endocrine adenocarcinomas in the small intestine through dedifferentiation of EC cells to intestinal stem cells, which are prone to transformation. However, these studies were limited by early death from tumors at other sites, leaving the potential for SI-NET development unclear over longer periods. To circumvent this time limited off target effect, the present study used intestinal enteroids from RPM mice to examine the effect of RB1 and Trp53 loss with or without gain of Myc function on EC cell derived tumors. Initial results confirmed previous in vivo induction of non-neuroendocrine adenomas. However, the addition of TNF-α to the enteroid media induced EC cell clusters in multiple crypts and well differentiated neuroendocrine tumor vs. carcinoma in the absence and presence of gain of Myc function, respectively. These findings suggest that TNF-α blocked EC cell dedifferentiation to ISCs, promoting their survival and expansion and shifting their fate from intestinal adenoma/carcinoma to a differentiated neuroendocrine tumor type. The present study thus highlights the crucial role of the microenvironment in influencing EC cell-derived tumorigenesis and provides insights into SI-NET development.

Myocardial infarction (MI) is a leading cause of mortality around the globe. Cardiac patches offer a promising tissue engineering approach to facilitate the natural regeneration of damaged cardiac tissue. In this research, electroactive cardiac patches composed of alginate-gelatin (Alg-Gel) were fabricated using a freeze-drying technique. The scaffolds were subsequently coated with varying concentrations of reduced graphene oxide (rGO) to improve cardiac performance. The samples were thoroughly characterized in terms of their physicochemical, morphological, and biological properties, including morphology, cell viability, and gene expression. Notably, an rGO concentration of 0.3% w/v significantly improved the viability of bone marrow-derived mesenchymal stem cells (BMSCs) and human umbilical vein endothelial cells (HUVECs), while upregulating the expression of Connexin 43 (Conx43), tryptophan transporter 2 (TrpT-2), and actinin 4 (Actn4), all of which are critical for cardiac cell function. To evaluate therapeutic efficacy in vivo, Alg-Gel-rGO scaffolds seeded with BMSCs were implanted into the infarcted area of a rat model of MI. Echocardiographic, histological, and immunohistochemical analyses demonstrated that the prepared patches improved cardiac function, reduced scar thickness, and promoted angiogenesis, thereby supporting cardiac tissue repair. These findings suggest that Alg-Gel-rGO scaffolds hold significant potential for regenerating damaged myocardial tissue and enhancing post-MI recovery, representing a viable strategy in cardiac tissue engineering.