Duchenne muscular dystrophy (DMD) is a debilitating disease characterized by progressive muscle-wasting and a lack of effective therapy. Although the application of GsMTx4 has been shown to reduce muscle mass loss in dystrophic mice, the mechanism of action remains unclear.

Fine particulate matter (PM2.5) exposure significantly exacerbates respiratory morbidity, particularly in asthmatic individuals, highlighting an urgent need for effective therapeutic interventions. In this study, we evaluated the therapeutic potential and underlying mechanisms of human dental pulp stem cells (hDPSCs), a promising mesenchymal stem cell population, in mitigating airway inflammation in mice with PM2.5-induced asthma exacerbation.

Vitiligo is an acquired skin depigmentation disorder often accompanied by leukoderma and leukotrichia. Half of vitiligo patients experience episodes of stress.

The subcellular localization of a protein is important for its function, and its mislocalization is linked to numerous diseases. Existing datasets capture limited pairs of proteins and cell lines, and existing protein localization prediction models either miss cell-type specificity or cannot generalize to unseen proteins. Here we present a method for Prediction of Unseen Proteins' Subcellular localization (PUPS). PUPS combines a protein language model and an image inpainting model to utilize both protein sequence and cellular images. We demonstrate that the protein sequence input enables generalization to unseen proteins, and the cellular image input captures single-cell variability, enabling cell-type-specific predictions. Experimental validation shows that PUPS can predict protein localization in newly performed experiments outside the Human Protein Atlas used for training. Collectively, PUPS provides a framework for predicting differential protein localization across cell lines and single cells within a cell line, including changes in protein localization driven by mutations.

Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) are a potential off-the-shelf product for acute ischemic stroke. This study explored the underlying mechanism of Cytopeutics® hUC-MSCs (Neuroncell-EX) as well as its feasibility and efficacy at two different doses: 2 × 106 cells per rat and 4 × 106 cells/rat in middle cerebral artery occlusion (MCAO) ischemic stroke model for 28 d. Modified neurological severity score (mNSS) and rotarod tests were evaluated at days 1, 4, 7, and 14. Transforming growth factor-beta 1 (TGF-β1), interleukin-1 receptor antagonist (IL-1Ra), and vascular endothelial growth factor (VEGF) were evaluated by enzyme-linked immunosorbent assay (ELISA) at days 4 and 28. Immunohistochemistry expression of aquaporin-4 (AQP4) and neuronal protein marker (NeuN) were performed at days 4 and 28, respectively. Both doses of Neuroncell-EX showed significant lower mNSS scores at days 7 and 14 compared to stroke control. Both Neuroncell-EX groups showed significant longer latency time at day 7, with only 4 × 10⁶ cells/rat group having significant longer time at day 14 than stroke control. At both time points, the 2 × 10⁶ cells/rat group had significantly higher TGF-β1 and IL-1Ra levels, with significantly increased TGF-β1 only observed in 4 × 10⁶ cells/rat group at day 4 compared to stroke control. The VEGF levels were significantly lower at day 4 but then significantly increased at day 28 in both Neuroncell-EX groups than stroke control. AQP4 expression was significantly higher in stroke control compared to healthy control at day 4. Both doses of Neuroncell-EX showed significantly higher NeuN expression compared to stroke control at day 28. There is a weak correlation between TGF-β1 with VEGF and inversely with AQP4. These results suggest that Neuroncell-EX is feasible and effective in promoting functional recovery and neuroprotection in ischemic rats, potentially through immunomodulation, angiogenesis, and neurogenesis mechanisms.

Whether one or two agents added to post-transplant cyclophosphamide (PTCy) are needed in HLA-matched allogeneic hematopoietic stem cell transplantation (allo-HSCT) with peripheral blood stem cells (PBSC) is debated. We retrospectively compared PTCy in association with a calcineurin inhibitor (PTCy+CNI) or with a CNI plus mycophenolate mofetil (PTCy+CNI+MMF) in adult patients transplanted for acute myeloid leukemia in first complete remission and receiving PBSC in the period from 2010 to 2020. Propensity score matching was performed using exact matching for donor type (related or unrelated) and the nearest neighbor for other variables (i.e. age, adverse cytogenetics, Karnofsky performance status, patient and donor cytomegalovirus serology, conditioning intensity). Each group comprised 146 patients, with 63% in total undergoing matched unrelated-allo-HSCT. Median follow up was longer for PTCy+CNI (36 [IQR 31-39] months versus 25 [IQR 19-30] months for PTCy+CNI+MMF, p < 0.01). At 2 years, PTCy+CNI was associated with a higher incidence of extensive chronic GVHD (16% [95% CI 10-22] versus 6% [95% CI 3-12] for PTCy+CNI+MMF, p < 0.03) while no differences were observed for all the other transplant outcomes. Addition of MMF to PTCy and CNI may help to prevent extensive chronic GVHD in HLA-matched allo-HSCT with PBSC.

Current methods for producing cardiomyocytes from human induced pluripotent stem cells (hiPSCs) using 2D monolayer differentiation are often hampered by batch-to-batch variability and inefficient purification processes. Here, we introduce CM-AI, a novel artificial intelligence-guided laser cell processing platform designed for rapid, label-free purification of hiPSC-derived cardiomyocytes (hiPSC-CMs). This approach significantly reduces processing time without the need for chronic metabolic selection or antibody-based sorting. By integrating real-time cellular morphology analysis and targeted laser ablation, CM-AI selectively removes non-cardiomyocyte populations with high precision. This streamlined process preserves cardiomyocyte viability and function, offering a scalable and efficient solution for cardiac regenerative medicine, disease modeling, and drug discovery.

Hypomethylating agents are frontline therapies for myelodysplastic neoplasms (MDS), yet clinical responses remain unpredictable. We conducted a phase 2 trial comparing injectable and oral azacitidine (AZA) administered over one or three weeks per four-week cycle, with the primary objective of investigating whether response is linked to in vivo drug incorporation or DNA hypomethylation. Our findings show that injection results in higher drug incorporation, but lower DNA demethylation per cycle, while global DNA methylation levels in mononuclear cells are comparable between responders and non-responders. However, hematopoietic stem and progenitor cells (HSPCs) from responders exhibit distinct baseline and early treatment-induced CpG methylation changes at regulatory regions linked to tissue patterning, cell migration, and myeloid differentiation. By cycle six-when clinical responses typically emerge-further differential hypomethylation in responder HSPCs suggests marrow adaptation as a driver of improved hematopoiesis. These findings indicate that intrinsic baseline and early drug-induced epigenetic differences in HSPCs may underlie the variable clinical response to AZA in MDS.

Psychological stress has profound impacts on the gastrointestinal tract via the brain‒gut axis. However, its effects on intestinal stem cells (ISCs) and the resulting implication for intestinal homeostasis remain poorly understood. Here, we observed a notable reduction in both the quantity and proliferative capacity of ISCs under chronic stress conditions, driven by elevated levels of corticosterone resulting from activation of the hypothalamic‒pituitary‒adrenal (HPA) axis. Mechanistically, corticosterone directly interacts with its receptor, nuclear receptor subfamily 3 group c member 1 (NR3C1), leading to increased expression of FKBP prolyl isomerase 5 (FKBP5) in ISCs. Subsequently, FKBP5 negatively regulates AKT activation by facilitating its dephosphorylation at Ser473, ultimately enhancing nuclear translocation of forkhead box O (FoxO) and inhibiting ISC proliferative activity. Consequently, ISC dysfunction contributes to the stress-driven exacerbation of DSS-induced colitis. Collectively, these findings reveal an intrinsic brain-to-gut regulatory pathway whereby psychological stress impairs ISC activity via corticosterone elevation, providing a mechanistic explanation for stress-enhanced susceptibility to colitis.

Doxorubicin (DOX) is the most effective chemotherapeutic for breast cancer, but it is usually associated with severe cardiotoxicity. Further investigation to alleviate its side effects is essential. The present study investigated the mechanism of the cross-organ communication between tumors and the heart and potential intervention targets. Morphological bubble-like protrusions were observed in both adult murine ventricular cardiomyocytes (AMVCs) and human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) cocultured with breast cancer cells (BCCs), along with elevated expression of pyroptosis-related proteins. Exosomes (EXOs) from DOX-treated BCCs aggravated DOX-induced cardiotoxicity (DOXIC) in an orthotopic mouse model of breast cancer. Blocking miRNAs by knocking down Rab27a or inhibiting the release of EXOs in cancer tissue by Dicer enzyme knockout attenuated this additional injury effect. Exosomal miRNA sequencing revealed that miR-216a-5p is especially upregulated in EXOs from DOX-induced BCCs. Mechanistically, miR-216a-5p was upregulated by enhanced transcription mediated by DOX-induced AMP-dependent transcription factor 3 (ATF3) and packaged into EXOs by splicing factor 3b subunit 4 (SF3B4) in BCCs. Itchy E3 ubiquitin-protein ligase (ITCH) was identified as a novel downstream target mRNA of miR-216a-5p. ITCH negatively mediated thioredoxin-interacting protein (TXNIP) ubiquitination to activate the NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) inflammasome pathway, ultimately leading to cardiomyocyte pyroptosis. Our findings revealed novel cross-organ pathogenic communication between breast cancer and the heart through the exosomal miR-216a-5p-mediated ITCH/TXNIP/NLRP3 pathway, which drives cardiomyocyte pyroptosis. These findings suggest that targeting myocardial miR-216a-5p or blocking harmful EXOs from breast cancer is a potential therapeutic strategy for alleviating DOXIC.

X-linked inhibitor of apoptosis protein (XIAP) deficiency is a congenital immunodeficiency disorder characterized by increased susceptibility to Epstein-Barr virus (EBV) infection and is frequently associated with hemophagocytic lymphohistiocytosis (HLH).

Lymphoma, a clonal malignancy from lymphocytes, includes diverse subtypes requiring distinct immunohistochemical stains for accurate diagnosis. Limited biopsy specimens often restrict the use of multiple stains, complicating diagnostic workflows. Lymphomas are typically classified into B-cell and T-cell types, each with specific markers. This study represents the first feasibility study in deploying deep learning models for B- and T-cell lymphoma classification on histopathological images. We analyzed 1510 H&E-stained sections (750 B-cell, 760 T-cell) with CNN models (Xception, NASNetL, ResNet50, EfficientNet), enhanced by Convolutional Block Attention Modules (CBAMs). All models demonstrated strong classification capabilities, with EfficientNet achieving the highest accuracy at 91.5% and the best precision at 91.9%, while Xception performed the best recall at 91.5%. Furthermore, the deep learning models significantly outperformed human pathologists in classification accuracy and inference speed, processing images in milliseconds compared to the several seconds required for manual diagnosis. These findings underscore the effectiveness of advanced CNN models in improving diagnostic precision while reducing dependency on manual staining and interpretation, and the integration of AI-driven classification can provide valuable support for pathologists.

Endometrial cancer (ECa) is a prevalent gynecological malignancy, with treatment often hindered by metastasis and recurrence driven by cancer stem-like cells. While circular RNAs (circRNAs) are well known for their cytoplasmic roles as microRNA sponges, their nuclear functions remain largely unexplored. This study investigates nuclear circRNAs and their roles in regulating cancer stem-like properties in ECa.

Photobiomodulation (PBM) uses light to stimulate cellular responses and has the potential to enhance cartilage regeneration through mesenchymal stem cell (MSC) differentiation. This study investigates the chondrogenic potential of adipose-derived MSCs (ADMSCs) under PBM using Red (630 nm, 72.7 mW) and Red/NIR (630 nm and 845 nm, 59.7 mW) organic light-emitting diodes (OLEDs), with ADMSCs exposed to two distinct OLED panels (10.85 cm2 surface area each). These wavelengths correspond to the absorption peaks of cytochrome c oxidase (CCO), a key mitochondrial enzyme involved in PBM-induced cellular bioenergetics. Cells were cultured in 24-well cell culture plate, receiving irradiance of 3.6 mW/cm2 and 2.74 mW/cm2, respectively. Exposure durations were 4.5, 14, and 23 minutes for Red OLED and 6, 18, and 30 minutes for Red/NIR OLED, delivering energy doses of 1, 3, and 5 J/cm² at 72-hour intervals. While Red/NIR OLED irradiation significantly enhanced ADMSC proliferation, it did not improve chondrogenic differentiation. In contrast, Red OLEDs consistently outperformed Red/NIR OLEDs as evidenced by stronger Safranin O staining, elevated collagen type II expression, and enhanced glycosaminoglycan (GAG) deposition via Alcian Blue staining. These findings underscore the superior efficacy of Red OLED-mediated PBM in promoting ADMSC differentiation toward the chondrogenic lineage and highlight the critical role of wavelength selection for PBM-based cartilage repair. Further exploration of the underlying mechanisms and optimization of PBM parameters is needed for improved clinical efficacies in tissue engineering and regenerative medicine.

Traumatic brain injury (TBI) is one of the primary causes of long-term brain disabilities among military personnel and civilians, regardless of gender. A plethora of secondary events are triggered by a primary brain insult, increasing the complexity of TBI. One of the most affected brain regions is the hippocampus, where neurogenesis occurs throughout life due to the presence of neural stem cells (NSC). Preclinical models have been extensively used to better understand TBI and develop effective treatments. Among these, rapid stretch injury has been used to mimic the effect of mechanical stress produced by a TBI on neurons and glia in vitro. In this study, we aimed to determine the impact of rapid stretch on the viability, proliferation, and differentiation of NSC isolated from rat hippocampus (Hipp-NSC) and to determine the role of the stretch-activated ion channel Piezo-1 in modulating their response to mechanical stress. We found that while rapid stretch (30 and 50 PSI) reduced Hipp-NSC viability (measured as a function of LDH release), it did not change their proliferation and differentiation potentials. Interestingly, rapid stretch in the presence of a selective Piezo-1 inhibitor, GsMTx4, or Piezo1 targeting siRNA, directed Hipp-NSC differentiation toward a neurogenic lineage. Additionally, we found that inhibiting Piezo1 with the addition of a rapid stretch injury increased the expression of miRNAs known to regulate neurogenesis. This work uses a novel approach for studying the effect of mechanical stress on NSC in vitro and points to the critical role the stretch-activated ion channel Piezo-1 has in modulating the impact of TBI on hippocampal neurogenesis.

Corneal repair using mesenchymal stem cell therapy faces challenges due to long-term cell survival issues. Here, we design cerium oxide with gold single-atom-based nanorobots (CeSAN-bots) for treating corneal damage in a synergistic combination with stem cells. Powered by glucose, CeSAN-bots exhibit enhanced diffusion and active motion due to the cascade reaction catalyzed by gold and cerium oxide. CeSAN-bots demonstrate a two-fold increase in cellular uptake efficiency into mesenchymal stem cells compared to passive uptake. CeSAN-bots possess intrinsic antioxidant and immunomodulatory properties, promoting corneal regeneration. Validation in a mouse corneal alkali burn model reveals an improvement in corneal clarity restoration when stem cells are incorporated with CeSAN-bots. This work presents a strategy for developing glucose-driven, enzyme-free, single-atom-based ultrasmall nanorobots with promising applications in targeted intracellular delivery in diverse biological environments.

Central trained immunity, induced via reprogramming of hematopoietic stem and progenitor cells (HSPCs), mediates sustained heightened responsiveness of mature myeloid cells to secondary challenges. We have previously demonstrated that HSPCs use TLR2 and Dectin-1 to sense Candida albicans to induce the production of trained monocytes/macrophages to fight against secondary infection. Neutrophils play an important role in innate immunity and are critical for clearance of C. albicans. In this work, we used an in vitro model of mouse HSPC differentiation to investigate the functional phenotype of neutrophils derived from HSPCs exposed to various PAMPs and C. albicans cells. We found that neutrophils derived from HSPCs stimulated by a TLR2 agonist exhibit reduced inflammatory cytokine production (tolerized neutrophils) whereas neutrophils generated from a Dectin-1 agonist or C. albicans stimulated HSPCs produce higher amounts of cytokines (trained neutrophils). We further demonstrated that a transient exposure of HSPCs to live C. albicans cells is sufficient to induce a trained phenotype of the neutrophils they produce in a Dectin-1- and TLR2-dependent manner. These trained neutrophils exhibited higher phagocytosis and microbicidal capacity than control neutrophils. Additionally, their adoptive transfer was sufficient to reduce fungal burden during invasive candidiasis. Mechanistically, we demonstrated that trained neutrophils use mitochondrial ROS (mtROS) to enhance their ability to kill C. albicans cells, as they produce higher amounts of mtROS and scavenging mtROS with MitoTEMPO attenuated their yeast-killing ability to match that of control neutrophils. Altogether, these data suggest that infection-experienced HSPCs contribute to trained immunity by providing a source of trained neutrophils with enhanced antimicrobial activity which may confer prolonged protection from infection. The tailored manipulation of this mechanism might offer new therapeutic strategies for controlling fungal infections by harnessing neutrophils.