Tissue repair is increasingly recognized as a systemic process influenced by age-associated changes beyond the injured organ itself. Clonal hematopoiesis of indeterminate potential (CHIP), a common consequence of somatic evolution in hematopoietic stem cells, has been linked to inflammatory disorders, yet whether it directly regulates tissue remodeling remains unclear. Here, we integrate population genomics, preclinical models, and human lung analyses to examine the role of CHIP in fibrotic lung disease. In large cohorts, idiopathic pulmonary fibrosis (IPF) was associated with a distinct CHIP mutational spectrum enriched for non- DNMT3A variants and for larger mutant clones. In mouse models, hematopoietic mutations exacerbated bleomycin-induced fibrosis and reprogrammed macrophages toward inflammatory, profibrotic states, including expansion of a distinct, injury-responsive SPP1 + population conserved in human disease. CHIP-associated macrophages were sufficient to directly promote fibroblast activation and alter epithelial differentiation, linking hematopoietic genotype to parenchymal remodeling. Consistently, a CHIP-derived macrophage transcriptional signature predicted adverse outcomes in independent IPF cohorts. Notably, immune and epithelial alterations were detectable even in the absence of overt injury, indicating that CHIP establishes a primed tissue environment permissive for maladaptive repair. Together, these findings identify clonal hematopoiesis as a systemic regulator of tissue repair and demonstrate that somatic evolution in blood can actively instruct organ remodeling through immune-parenchymal interactions. This framework supports the possibility that disease-associated selective pressures may shape clonal architecture with functional consequences for organ health.

Adult stem cells are thought to drive the regenerative potential of the endometrium and contribute to the pathogenesis of endometriosis, however, their identity and defining features remain to be characterized. Here, we used in vivo and in vitro approaches to demonstrate that cells with high aldehyde dehydrogenase 1 activity (ALDH HI cells) were long lived progenitors in the endometrial epithelium with a higher organoid formation capacity, long-term passaging potential, and stemness gene signatures. Using lineage tracing with an Aldh1a1cre/ERT2; ROSA26tdTomato reporter mouse, Aldh1a1+ cells expanded during postnatal development, estrus cycling, and following post-partum repair. In response to ovariectomy or exogenous estradiol, we found that ALDH1A1 + cells localized to glandular crypts of the endometrium or throughout the luminal epithelium, respectively, indicating that their spatial localization is hormone sensitive. Functionally, we found that selective ablation of ALDH1A1 + cells in Aldh1a1cre/ERT2; ROSA26- DTR flox/flox mice decreased endometrial gland number and FOXA2 expression . These findings were recapitulated in the human endometrium, where endometrial epithelial organoids with high ALDH activity (ALDH HI cells) showed a higher organoid formation capacity than ALDH LO cells and displayed unique transcriptomes with fewer luminal-like ciliated cells. Overall, our studies indicate that ALDH1A1 + cells are hormone-sensitive adult stem cells in the endometrium with regenerative potential that are critical for endometrial development and function.

Directed differentiation of pluripotent stem cells (PSCs) into pancreatic islets is a cornerstone strategy for diabetes cell therapy. This process relies on growth factor-driven activation of core transcriptional regulators, notably PDX1 and NGN3, to restrict the multi-lineage potential of definitive endoderm to pancreatic progenitors and endocrine cell types. Yet differentiation efficiency and lineage fidelity vary markedly across PSC lines. Here, we demonstrate that a dominant constraint is persistent Polycomb Repressive Complex 2 (PRC2)-mediated epigenetic repression at the PDX1 and NGN3 loci, limiting endocrine specification despite inductive signaling. To directly test whether chromatin states at the PDX1 and NGN3 loci gate developmental competence, we deployed a computationally engineered epigenetic effector (EBdCas9) to transiently and sequentially remove H3K27me3 at those loci during defined developmental windows. Targeted epigenetic resolution robustly enhanced endocrine lineage commitment and accelerated β-cell differentiation across genetically diverse PSC lines. In contrast, direct transcriptional activation with VP64dCas9 increased PDX1 and NGN3 expression but did not improve differentiation outcomes. Integrated cell population studies and genome-wide chromatin and transcriptomic analyses reveal that PRC2-targeted remodeling preferentially activates endocrine gene networks while limiting progenitor expansion and lineage-inappropriate programs. These findings establish that gene-targeted manipulation of PRC2-mediated repression at PDX1 and NGN3 can be used to control cell lineage competence. Collectively, our study reframes variability in PSC differentiation as a failure of epigenetic resolution rather than transcriptional insufficiency and introduces locus-specific chromatin remodeling as a generalizable strategy to enforce developmental fidelity.

Tumor cells adapt to hypoxia by releasing hiTDExs enriched with bioactive molecules that modulate endothelial behavior and promote tumor progression. This study aimed to characterize how hypoxia-induced HNSCC exosomes reshape the endothelial secretome and contribute to metastatic potential.

Dental follicle stem cells (DFSCs) originate from the dental follicle during tooth development and possess multilineage differentiation potential, contributing to periodontal tissue regeneration, bone repair, and immunomodulation. This review highlights the recent advances in the application of DFSCs and biological scaffolds for regenerative medicine, with a focus on oral and craniofacial tissue. DFSCs exhibit key advantages for regenerative therapies, including high accessibility, robust self-renewal capacity, and multipotent differentiation potential, enabling their differentiation into odontogenic (dentin- and enamel-forming), osteogenic, and fibroblastic lineages. We discuss the embryonic origin of DFSCS and their unique ability to maintain stable cellular properties in long-term in vitro culture. Importantly, DFSCs play a pivotal role in tooth morphogenesis, periodontal tissue formation, and craniofacial bone regeneration, making them promising for functional oral tissue restoration. A critical aspect of DFSC-based regeneration is the integration with bioactive scaffolds, which provide structural support, promote cell adhesion, proliferation, and differentiation, and facilitate vascularization. We analyze how scaffold properties, such as biodegradability, porosity, and permeability, influence DFSC behavior and therapeutic outcomes. Finally, we explore future challenges and opportunities in optimizing DFSC-scaffold interaction, emphasizing advancements in biomaterial design and emerging bioengineering technologies. Preliminary evidence suggests that integrating DFSCs with engineered scaffold systems may offer potential benefits for personalized regenerative therapies, though further validation is required before clinical translation. Such approaches could contribute to advancing tooth and craniofacial reconstruction strategies. This review consolidates existing insights and explores potential avenues for future research to support advancements in DFSC-based regenerative medicine.

Virus-specific CD4+ T cells are essential for coordinating adaptive immunity during infection, but their differentiation and maintenance in chronic infection remain unclear. Using human hepatitis C virus (HCV) infection as a model, we assessed the determinants of virus-specific CD4+ T cell immunity in acute, spontaneously cleared, chronic, and therapeutically cured infections. During acute infection, multiple subsets of progenitor CD4+ T cells emerged, including subsets that are also found in chronic infection. In chronic infection, stem-like Bcl-2+ CD4+ T cells and T-bet+ effector CD4+ T cells existed in a progenitor/progeny relationship. Following therapy-mediated HCV cure, these cells retained their chronic signature but formed a stable memory pool that persisted for years and was distinct from HCV-specific CD4+ T cell memory after spontaneous clearance. Collectively, our findings highlight differences in CD4+ T cell fates that depend on infection outcomes and reveal common principles of CD4+ and exhausted CD8+ T cell maintenance during and after chronic infection.

The cellular, immunogenetic, and antigenic factors affecting the breadth of viral antigen variants recognized by human antibody responses are poorly defined. We developed highly multiplexed panels of DNA-tagged SARS-CoV-2 antigens from up to 20 viral variants to label and sort 6,262 antigen-binding circulating B cells from previously naive mRNA vaccinees or infected patients, and from deceased organ donor lymphoid tissues, to enable antigen receptor and transcriptome sequencing. Atypical B cells and a subset of class-switched memory cells with evidence of recent germinal center exposure were enriched for antigen binding. In contrast to atypical B cells, post-germinal center B cells showed progressively increasing variant binding breadth and somatic hypermutation over time. Vaccination, compared with infection, preferentially stimulated B cells expressing antibodies with inherently high antigen-binding breadth. This large-scale analysis reveals key determinants of antigen-binding breadth, critical for understanding responses to viral infection and guiding vaccine development against rapidly mutating viruses.

Regulatory T cells (Tregs), winners of the 2025 Nobel Prize in Physiology or Medicine, emerged as an unsung giant of peripheral immune tolerance after allogeneic hematopoietic stem cell transplantation (allo-HSCT). These cells exert influence across four interconnected immunological axes encompassing graft-versus-leukemia (GVL), graft-versus-host disease (GVHD), relapse, and viral infection. This correspondence highlights key advancements in Treg-based interventions in allo-HSCT, ranging from basic research to clinical applications, as presented at the ASH 2025 Annual Meeting. Accordingly, Tregs are used as GVHD prophylaxis via the Orca-T platform and as a therapeutic strategy for steroid-refractory (SR)-GVHD patients. Experimental studies further pave the way to address existing limitations toward next-generation Treg-based cell therapy in allo-HSCT.

Reprogramming tumor-associated macrophages (TAMs) from the pro-tumoral M2-like state to the immunostimulatory M1-like phenotype has emerged as a promising strategy for tumor therapy. However, most M2-like TAMs are preferentially located in hypoxic regions of the tumor, which are poorly accessible to many advanced drug delivery systems, posing a significant challenge to effective TAM reprogramming. Here, leveraging the tropism of facultative anaerobic bacteria to localize and propagate in the hypoxic tumor, an optogenetically engineered Escherichia coli Nissle 1917 strain conjugated with murine STING agonist (EcNflaB@UPD) was developed for cancer-specific immunotherapy. Upon near-infrared light illumination, the blue and UV emissions from upconversion nanoparticles (UCNPs) simultaneously activate the expression of Toll-like receptor (TLR) agonist, flaB, from EcNflaB, and the release of photocaged murine STING agonist, DMXAA, respectively. This spatiotemporally synchronized dual release ensures co-localized STING and TLR5 agonists inside the hypoxic niche, repolarizing TAMs from the M2 to the M1 phenotype via synergistic TLR5-MAPK1-NF-κB and STING-NF-κB signaling. The polarization of TAMs enhances their antigen-presenting capacity and, more importantly, activates the cytotoxic, stem-like and memory CD8+ T cells responses. This subsequently inhibits tumor growth, relapse, and metastasis in the murine 4T1 tumor model. Collectively, our work introduces the bacteria-based system that uses near-infrared light to dual-release immunotherapeutics for systemic anti-tumor immunity, opening new avenues for precise and effective cancer immunotherapy.

Reprogrammable Adenosine Deaminase Acting on RNA (ADAR) Sensors (RADARS) control RNA translation in mammalian cells, allowing for noninvasive sensing or perturbation of specific cell types based on transcriptional signatures. Upon base-pairing between a target RNA and a sensor RNA, RADARS leverages ADAR to edit a premature stop codon upstream of a gene of interest, thereby releasing translation of the desired cargo. These design principles enable sequence programmability, allowing RADARS to adapt more easily to new contexts than existing tools for targeting cell types. We describe a detailed protocol for performing experiments with RADARS, including designing, cloning and validating RADARS constructs targeting a transcript of interest. RADARS guide sequences can be designed with an intuitive web interface and cloned into existing constructs for downstream applications including imaging, sorting and sequencing. We outline recommendations for cargo choice, sensor design and ADAR system selection, enabling users to choose the best workflow depending on the desired application. Beginning with sensor design, the selection of top-performing RADARS guides can be completed in ~2 weeks, followed by a desired use case. Convenient engineering and application of RADARS for various applications enable the design and execution of various cell-targeting experiments.

Growing evidence implicates early metabolic dysfunctions in retinal ganglion cells (RGCs) as a contributor to both high- and normal-tension glaucoma, yet no approved therapy directly protects RGCs to preserve vision. We aimed at identifying a safe, druggable neuroprotective strategy that restores RGC metabolic homeostasis for glaucoma therapy.

Successful autologous stem cell transplantation (ASCT) in newly diagnosed multiple myeloma (NDMM) patients relies on the efficient mobilization of hematopoietic stem cells following induction therapy. While the efficacy of etoposide for stem cell mobilization has been demonstrated in numerous studies, a randomized comparison of the efficacy of cyclophosphamide versus etoposide has previously been lacking. This randomized, open-label, multicenter trial enrolled NDMM patients eligible for ASCT. The inclusion criteria were patients with a diagnosis of NDMM who required stem cell mobilization prior to ASCT. Patients were randomly assigned to receive either high-dose etoposide (VP16; 1.2 g/m2) or high-dose cyclophosphamide (CTX; 3.0 g/m2) before mobilization. Granulocyte colony-stimulating factor (G-CSF) was administered after chemotherapy to promote stem cell mobilization. The primary endpoint was the proportion of patients achieving CD34 + cell counts ≥ 2 × 10⁶/kg and ≥ 5 × 10⁶/kg. A total of 62 patients were enrolled, with 31 patients in each group. The VP16 group significantly outperformed the CTX group in CD34 + cell collection across all thresholds: ≥2 × 10⁶/kg (100% vs. 77%, p = 0.011), ≥ 5 × 10⁶/kg (90% vs. 55%, p = 0.002), and ≥ 8 × 10⁶/kg (71% vs. 32.3%, p = 0.023). The VP16 group also showed superior success rates in the first apheresis session and achieved higher CD34 + percentages in the collection. Additionally, the VP16 group required fewer apheresis sessions, fewer platelet transfusions, and experienced less nausea during the mobilization period. High-dose etoposide (1.2 g/m2) demonstrated superior efficacy and safety compared to high-dose cyclophosphamide (3.0 g/m2) for stem cell mobilization in NDMM patients. Based on these findings, etoposide may be considered a more effective and safer option for stem cell mobilization in clinical practice.The clinical trial was registered on 24/08/2022 (clinical trial identifier NCT05517213).

In this study, we demonstrate that cytoskeletal state at the onset of directed differentiation impacts the exit of human pluripotent stem cells (hPSCs) from pluripotency and downstream lineage specification. In particular, depolymerizing F-actin with latrunculin A (latA) during the first 24 h of definitive endoderm formation facilitates efficient loss of pluripotency and alters Activin/Nodal, BMP, c-Jun, and WNT signaling dynamics. These signaling changes influence downstream patterning of the gut tube, leading to improved pancreatic progenitor identity and decreased expression of markers associated with other endodermal lineages. Continued differentiation generates islets containing a higher percentage of β cells that exhibit improved maturation, insulin secretion, and ability to reverse hyperglycemia. Furthermore, this latA treatment reduces enterochromaffin cells in the final cell population and corrects differentiations from hPSC lines that otherwise fail to consistently produce pancreatic islets, highlighting the importance of cytoskeletal signaling at the onset of directed differentiation.

Glioblastoma (GBM) is an aggressive primary brain tumor marked by a poor prognosis and limited effectiveness of current therapies, which are often accompanied by substantial recurrence rates. To address this challenge, we developed BM03, an engineered mesenchymal stem cell (MSC) line specifically designed for glioblastoma therapy. BM03 is structured to enhance migration to GBM sites and deliver targeted, multimodal gene-based therapies.

Compared to the general population, individuals with Neurofibromatosis Type 1 have a 50-fold higher risk of developing a high-grade glioma (HGG) in their lifetime. Despite improved understanding of the molecular and cellular drivers of these neoplasms, we have yet to translate this knowledge into therapies that improve overall survival. One limitation has been the paucity of in vivo models for drug testing within this population. We generated 3 distinct glioma stem cell lines from high-grade gliomas arising in mice with Nf1 and Trp53 mutations in cis (NPcis) and characterized the allografts resulting from one Nf1-glioma stem cell line (Nf1-HGG17) by single cell and single nuclear RNA-seq. Because our cell lines are grown in stem cell media, there is an inherent reduction of the differentiation states present in primary HGG, with only oligodendrocyte precursor-like and neural-like cells identified. However, orthotopic allografts of Nf1-HGG17 regained the other differentiation states typically observed in HGG and GBM (neuronal progenitor cell-like and astrocyte-like). About half of neoplastic cells cluster separately from these classical groups and highly express genes of the antigen presentation machinery, including Cd74, which we also observed in patient samples. Notably, heterozygosity of Nf1 within the tumor microenvironment does not cause marked changes in the immune tumor microenvironment, gene expression, or differentiation states of neoplastic cells. These data indicate that allografted HGG lines from NPcis mice are an effective model of NF1-HGG, mimicking the complexity observed in human HGG, that can be used for larger scale in vivo drug screening and evaluation.

Wound healing is complex, and hard-to-heal (chronic) wounds pose significant treatment challenges, especially in adults. Micrografts (MGs) are emerging as a promising treatment for wounds refractory to conventional approaches. MG involves transplanting a stem cell suspension to the wound to promote healing. Scientific studies on MG are increasing; however, a systematic review is needed for a comprehensive understanding of its efficacy.

The amygdala paralaminar nuclei (PL) contain immature excitatory neurons that develop on a delayed timeline from birth to adulthood and are more prominent in the amygdala of humans and other primates than in rodents. Whether this expansion is linked to brain complexity or is a feature of primates is unknown. We sought to identify the PL in the ferret (Mustela putorius furo), a small, gyrencephalic mammal that does not belong to the primate order. Here, we show that the amygdala of juvenile (P30-P67) and adult ferrets (>1 year) also contains a collection of immature excitatory neurons that express doublecortin (Dcx) and polysialylated neural cell adhesion molecule (Psa-Ncam). Similar to humans and mice, these immature neurons express Tbr1 and CoupTFII, but not FoxP2, which labels neighboring clusters of GABAergic cells in the intercalated nuclei. Ferret PL neurons extend into the ventral basolateral amygdala (BLA) and appear either in dense clusters surrounded by astroglia or as individual cells, and each subpopulation contains neurons with migratory morphology. This expansion of PL neurons into the amygdala is similar to what is seen in humans, but unlike in mice, where PL neurons are infrequent in the BLA. We compared these findings to the marmoset (Callithrix jacchus), a lissencephalic non-human primate, and the swine (Sus scrofa domesticus), a gyrencephalic mammal, and found immature neurons extending into the amygdala in both. Our study identifies the PL region of the ferret amygdala, which contains immature neurons with migratory features in juveniles and adults. Cross-species comparisons indicate that the expansion of PL neurons into the amygdala seen in primates with both high and low gyrencephalic indices has also occurred in species with gyrencephalic brains from different orders.

Chromosome karyotyping, particularly G-banding, is a fundamental diagnostic and prognostic tool for hematological malignancies, providing a genome-wide view of large-scale numerical and structural chromosomal abnormalities. Its clinical utility is paramount for disease classification, risk stratification, and the evaluation of hematopoietic stem cell transplantation (HSCT) across diseases such as acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), multiple myeloma (MM), and myelodysplastic syndromes (MDS). However, clinical challenges including low resolution and culture failure necessitate complementary advanced techniques. Fluorescence in situ hybridization (FISH) targets specific aberrations in non-dividing cells, while array comparative genomic hybridization (aCGH) and single-nucleotide polymorphism (SNP) arrays offer higher resolution for detecting cryptic copy number variations (CNVs) and copy-neutral loss of heterozygosity (CN-LOH). Furthermore, the modern diagnostic standard has evolved into a multi-omics approach that integrates morphology, flow cytometry, karyotyping, and next-generation sequencing (NGS). This comprehensive workflow significantly enhances diagnostic accuracy, refines risk stratification, and informs personalized therapeutic strategies. Clinically, karyotyping is essential for assessing cytogenetic remission, though it is less sensitive for minimal residual disease (MRD) detection than molecular methods. As emerging technologies such as optical genome mapping (OGM) demonstrate the potential to streamline these workflows, karyotyping continues to evolve, solidifying its indispensable role in the comprehensive management of hematologic cancers.