The olfactory epithelium possesses an adult stem cell population, the horizontal basal cells (HBCs), to permit lifelong tissue regeneration. Here, we show that HBCs exhibit asymmetric inheritance of histone H4 but not H2A-H2B during olfactory epithelium regeneration in mice. Primary HBC cultures further revealed asymmetric histone inheritance for H3 and H3.3. Upon mitotic exit, asymmetric histone inheritance correlates with asynchronous transcription re-initiation and differential enrichment of p63, a key transcription factor for HBC cell fate. Disruption of asymmetric histone inheritance abolishes these asymmetric cellular features and attenuates olfactory epithelium regeneration and smell behavior recovery. Single-cell RNA sequencing of paired HBC daughters in culture further supports asymmetric multilineage cell fate priming. Together, these findings reveal asymmetric histone inheritance in a mammalian adult stem cell lineage and highlight its biological significance in neural tissue regeneration and animal behavior.

Tissue-resident stem cells play an essential role in repairing barrier tissues subjected to frequent insults. However, the local cues that coordinate successful barrier repair or lead to tissue remodeling are largely unknown. Here we use murine models of airway injury, fate mapping, and null strains to identify a role for rare tuft epithelial cells in signaling to submucosal stem cells through the generation of cysteinyl leukotrienes (CysLTs) and activation of the CysLT receptor OXGR1. This results in mobilization of SOX9+ submucosal gland progenitors, aberrant repair of the surface airway epithelium, and durable features of airway remodeling including submucosal gland hyperplasia and collagen deposition. Remarkably, selective deletion of SOX9 from the airway stem compartment allows epithelial restoration and prevents tissue remodeling. These findings demonstrate a tuft cell- OXGR1- and SOX9- circuit that remodels the airway after injury and is detected in the human sinus mucosa.

DIAPH3 is a master regulator of the cytoskeleton with key roles in cell division. In the mouse brain, DIAPH3-deficient neural stem cells exhibit abnormalities in karyokinesis and cytokinesis, leading to cell cycle arrest, aneuploidy, and mitotic catastrophe. Here, we investigated the role of DIAPH3 in glioma genesis in mouse models. We selectively deleted the Diaph3 and Trp53 genes in the mouse cerebral cortex and thoroughly analyzed single (Diaph3 cKO and Trp53 cKO) and double (dcKO) conditional knockout mice. The tumors appeared earlier in dcKO than in Trp53 cKO mice, and this was associated with increased whole chromosome copy number alterations, endogenous DNA damage, and shorter survival of dcKO mice. We performed a comparative transcriptomic analysis prior to the onset of tumors and identified changes in cancer gene signatures specifically in dcKO, suggesting that the loss of DIAPH3 hastens the tumorigenic process. We isolated cancer stem-like cells and assessed their sensitivity to ionizing radiation and found that DIAPH3 regulates the resistance of glioma stem-like cells to irradiation. Our data suggest that DIAPH3 has a tumor-suppressor function and that its deficiency promotes aneuploidy and genome instability, accelerating tumorigenesis and leading to early onset of high-grade diffuse glioma with DNA damage, and resistance to ionizing radiation.

Brain inflammation leads to an increase in the amount of iron in brain tissue, but the source of the iron that could lead to accumulation remains unclear. Our recent in vitro study discovered that, in addition to the traditional pathway, iron can be released from the blood-brain barrier via extracellular vesicles (EVs). Herein, we investigated the impact of brain inflammation on iron release via EVs from the brain microvasculature (BMV). For this study, we induced brain inflammation in three-month-old C57BL/6 by intracerebroventricular injection of lipopolysaccharide (LPS,12μg/mice). For in vitro, we used human blood-brain barrier endothelial cells derived from human-induced pluripotent stem cells (hiPSCs). The LPS injection activated microglia and astrocytes as well as increased brain proinflammatory cytokines compared to the control mice. Furthermore, brain inflammation increased the iron levels in the brain parenchyma but decreased the iron levels in BMV. Brain inflammation was associated with the degradation of ferroportin (FPN1), an iron exporter, in the BMV. CD63, an EVs membrane protein, was increased in the BMV and associated with increased FTH1 release via EVs from BMVs to the brain. Moreover, brain inflammation decreased iron in BMV as evidenced by an increase in the transferrin receptor and decreased FTH1, suggestive of increased iron uptake. Pharmacological reduction of EVs by GW4869 reduced iron accumulation in the inflamed brain parenchyma compared to control mice. In summary, we have discovered a novel mechanism by which BMV-released EVs enriched with FTH1 and reveal a significant mechanism for brain iron accumulation during inflammation.

Cell therapy remains an attractive therapeutic option for the numerous genetic and non-genetic maladies affecting skeletal muscle. Since skeletal muscle is the largest tissue in the body, delivery has been notoriously challenging, but there have been significant advances, with several ongoing clinical trials of allogeneic and autologous cell transplantation aiming to replace diseased skeletal muscle with healthy and functional myofibers and muscle stem cells. Paracrine cellular approaches intended to enhance regeneration are also ongoing. In this review, we will provide an overview of the progress and current status of these different approaches, and discuss the forecast for future phases as well as the hurdles that need to be circumvented for the widespread application of cell therapy for skeletal muscle disorders.

Mycoplasma pneumoniae (MP) infection directly damages lung tissue and triggers dysregulated immune responses, yet the immunopathology underlying the severe pneumonia remains unknown. Here we analyzed cellular and molecular features in the peripheral blood and bronchoalveolar lavage fluid from patients to define the systemic immune landscape associates with severity of MP pneumonia (MPP). Cytokine and cellular profiling found severe MPP is associated with type I inflammation potentiated by γδ T and NK T cells. MPP patients were found to have autoantibodies targeting non-protein antigens such as dsDNA, phosphatidylserine and cardiolipin, and a broad panel of peptide antigen such as CUL4B, a core component of the ubiquitin ligase complex which is significantly enriched in severe MPP. Mechanistically, we identified elevated plasma cells correlating with increased autoantibody levels, as well as disease severity. These findings suggest a potential association between cellular immune responses and autoantibody generation, advancing our understanding of MPP pathogenesis and revealing novel potential therapeutic targets.

Sarcomas and carcinomas represent approximately 1% and 80% of all cancer diagnoses, respectively. Despite their very different prevalence, both tumor types share a critical dependence on mitochondrial functions for metabolic adaptation, survival and progression. Mitochondria act as cellular powerhouses by generating ATP through oxidative phosphorylation; however, their roles extend far beyond energy production. These organelles are central hubs of biosynthetic and catabolic pathways, including the tricarboxylic acid cycle, glutaminolysis, lipid metabolism, branched-chain amino acid catabolism and gluconeogenesis. Moreover, they play a key role in regulating various forms of programmed cell death, such as apoptosis, necroptosis, ferroptosis and pyroptosis. In this review, we provide a comprehensive overview on the contribution of mitochondria to tumor cell metabolism specifically in sarcomas and carcinomas. We describe how mitochondrial DNA-encoded proteins influence tumorigenesis and how mitochondria support cancer stem cell maintenance. We also discuss the therapeutic potential of targeting mitochondrial pathways, highlighting clinical trials and emerging strategies. The available evidence suggests that sarcoma cells might be more responsive to mitochondrial-targeted therapies due to their higher mitochondrial content and activity compared with carcinomas. Lastly, we bring some evidence of the involvement of mitochondria in the tumor microenvironment and discuss the implication of this finding for cancer immunotherapy. Altogether, these insights emphasize the importance of mitochondria as central regulators of cancer cell fate and promising therapeutic targets.

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.

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.

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.

Modulation of miRNA expression in glomerular cells is associated with renal disease. Here, we investigated the role of miR-93-5p in mitigating glomerular damage in Alport syndrome and whether the disease-modifying activity of extracellular vesicles from human amniotic fluid stem cells (hAFSC-EVs) is mediated by their miR-93-5p cargo. We identified downregulation of miR-93-5p specifically in glomerular endothelial cells in Alport syndrome along disease progression. Silencing of miR-93-5p in hAFSC-EVs changed the transcriptomic and proteomic profile, regulating EV disease-modifying activity. Compared with naive hAFSC-EVs, silenced hAFSC-EVs did not rescue glomerular endothelial function in vitro and did not restore kidney function in vivo. We established that hAFSC-EVs regulate VEGFR1 and VEGFR2 signaling by miR-93-5p cargo transfer, highlighting that miR-93-5p can restore glomerular endothelial cell biology. Spatial transcriptomics analysis of hAFSC-EV-injected kidneys showed that these EVs can reverse pathways altered during disease progression by stimulating proregenerative processes, specifically in the glomerulus, by regulating miR-93-5p targets. Alteration of glomerular endothelial cell transcriptomics and miR-93-5p targets was also confirmed in biopsies of patients with Alport syndrome using spatial molecular imaging. We demonstrated the critical role of miR-93-5p in glomerular endothelial cells and the capability of hAFSC-EVs to regulate miR-93-5p and its targets in Alport syndrome.

Limb-girdle muscular dystrophy R2 (LGMD R2) is an autosomal recessive disorder caused by dysferlin deficiency, leading to progressive muscle weakness and wasting. The lack of reliable clinical biomarkers has limited disease monitoring and therapeutic evaluation. Here, we identified Disabled-2 (DAB2) as a molecular and clinical indicator of disease state in LGMD R2. Transcriptomic profiling revealed a significant upregulation of DAB2 in induced pluripotent stem cell-derived (iPSC-derived) myotubes from patients, a finding validated in muscle biopsies from 14 dysferlin-deficient individuals and in dysferlin-deficient Bla/J mice, where DAB2 levels increased with disease progression. Importantly, AAV-mediated expression of full-length dysferlin restored DAB2 levels, supporting its value as a dynamic readout of disease activity for both disease monitoring and therapeutic response. Given the established role of DAB2 in clathrin-mediated endocytosis, particularly in LDL receptor internalization and cholesterol homeostasis, and the pathological lipid accumulation reported in LGMD R2, we investigated its contribution to lipid dysregulation. High DAB2 expression paralleled lipid deposition in patient muscles, iPSC-derived myotubes, and mouse tissue, whereas siRNA-mediated DAB2 knockdown reduced lipid accumulation in LGMD R2 myotubes. Collectively, these findings suggest that DAB2 functions as a mechanistic link between dysferlin deficiency, altered lipid handling, and disease severity, and they highlight its potential as a prognostic marker and therapeutic response measure for LGMD R2.

Common variable immunodeficiency (CVID) is the most prevalent symptomatic primary antibody deficiency. For unclear reasons, inflammatory complications, like gastrointestinal (GI) disease, occur in ~50% of CVID cases, worsening morbidity and mortality. NFKB1 variants are among the most frequent genetic variants in CVID. While effect of NFKB1 variants is not well understood, we previously found frameshift heterozygous NFKB1 variants to increase cytokines, monocytes, and inflammatory complications in CVID. In this report, we used induced pluripotent stem cell-derived (iPSC-derived) monocytes (iMONOs) with CRISPR/Cas9-mediated gene editing to study a heterozygous NFKB1 frameshift found in a patient with CVID with severe GI disease. The heterozygous NFKB1 variant similarly reduced NFKB1 protein in CVID patient- and healthy donor-derived iMONOs, but elevated LPS-induced IL-1β release and expression of inflammatory genes, including IL1B, IL6, TNF, and neutrophil chemoattractants, only in CVID patient iMONOs. CVID patient iMONOs also had elevations of IL-12, CCL4, and CCL12 unaffected by presence or absence of the NFKB1 variant. TNF antagonism improved the patient's GI disease, diminishing neutrophilic gastritis, circulating neutrophils, and the neutrophil chemoattractant CXCL1 in the blood. While the biology remains complex, our approach found heterozygous NFKB1 variant-induced inflammatory changes intensified in CVID iMONOs, corresponding with clinical response to TNF antagonism.

Pancreatic ductal adenocarcinoma (PDAC) is among the most lethal malignancies, characterized by aggressive biology, a dense fibrotic and immunosuppressive microenvironment, and profound resistance to standard therapies. Smart polymeric nanoparticles (SPNs), engineered to sense and respond to biological cues, present a transformative approach to overcome these barriers. This review highlights recent advances in SPNs tailored for PDAC, including systems designed to actively target tumor cells, cancer-associated fibroblasts (CAFs), and cancer stem cells (CSCs), thereby enhancing selective drug delivery and efficacy. SPNs also remodel the desmoplastic stroma or deliver matrix-modulating agents to improve tumor penetration. Furthermore, stimuli-responsive SPNs exploit the unique tumor microenvironment (TME) of PDAC, leveraging pH, hypoxia, or enzymatic triggers to achieve controlled, localized drug release. Beyond these strategies, SPNs have been developed to reprogram tumor immunity, modulate metabolic pathways, and enable precision gene therapy or combination treatments. Incorporating chronotherapy principles, future SPNs are capable of synchronizing drug release with circadian rhythms to maximize therapeutic windows while minimizing toxicity. Emerging concepts, such as integrating biosensors for real-time endogenous signal detection or applying AI-driven design to optimize SPN properties, underscore the future potential of these systems. Together, these multifaceted strategies position SPNs as a powerful platform to tackle the formidable challenges of PDAC and advance toward personalized cancer care.

Intracranial germ cell tumors (iGCTs) are categorized into germinomas (GEs) and non-germinomatous germ cell tumors (NGGCTs), which show divergent clinical outcomes with GEs having a significantly more favorable prognosis. This prognostic disparity suggests distinct biological characteristics, yet the association between tumor stemness and the host immune microenvironment, particularly in pre-treatment peripheral blood, remains poorly understood. Therefore, this study integrated multi-omics data, immune cell enrichment scores, and peripheral blood analyses to elucidate the relationship between cancer stemness indices and immune cells, especially neutrophils; and to evaluate its prognosis and treatment strategy in iGCTs.