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Mesenchymal stem cells (MSCs) have emerged as a promising therapeutic approach for degenerative diseases due to their ability to modulate disease progression through paracrine mechanisms. Among various MSC sources,placenta-derived MSCs (PD-MSCs) offer significant advantages,including high proliferation capacity,reduced senescence,and low immunogenicity,making them ideal for allogeneic applications. In this study,we investigated the therapeutic effects of PD-MSC transplantation in an estrogen-deficiency-induced osteoporosis mouse model. Mice were divided into three groups: a normal control group,a non-transplanted osteoporosis group,and a PD-MSC-transplanted group. Our findings demonstrated that PD-MSC transplantation significantly improved osteoporosis-related parameters,including increased femur weight,bone volume,bone mineral density,and calcium deposition. Additionally,estrogen levels were elevated,bone formation markers were upregulated,and bone resorption markers were downregulated. PD-MSCs also reduced inflammatory cytokine levels while enhancing anti-inflammatory factors. Notably,the BMP/SMAD signaling pathway,crucial for bone formation,was significantly upregulated. These results suggest that PD-MSC transplantation effectively restores bone homeostasis by inhibiting osteoclast activity,promoting osteogenesis,and modulating inflammation. This study provides strong evidence supporting the potential of PD-MSCs as a novel therapeutic strategy for osteoporosis,offering a regenerative and anti-inflammatory approach to bone disease management. View Publication

ObjectivesThis study evaluated Biodentine (BD) after modification with 2 wt% fluorinated graphene (FG).MethodsFG was prepared using the modified Hummers’ method,where sulfuric and phosphoric acids were added to fluorinated graphite and potassium permanganate. The mixture was heated,sifted,filtered,and centrifuged to obtain FG powder. Characterization was performed using XRD,FTIR,TEM,and SEM/EDX. PH was evaluated,And Ca And F ion release were assessed by inductively coupled plasma spectroscopy and ion chromatography,at days 1,14,And 28. Cell viability was performed using the MTT Assay on pulp stem cells,while ALP assay was evaluated by a spectrophotometer. Compressive strength was evaluated by a universal testing machine. Statistical analysis was performed on the data (p ≤ 0.05).ResultsGraphene and C-F bonds of FG were confirmed in XRD and FTIR,while nanosheets were detected in TEM. SEM/EDX showed more surface roughness in modified BD-FG. pH And Ca ion release results showed significantly higher values at day 1 for modified BD-FG,with significantly higher cumulative Ca ion release. Cell viability results showed no significant difference between modified And unmodified Biodentine at days 1 And 7; however,modified BD-FG showed significantly lower values at day 3. No significant difference was observed between the two groups in ALP,while the BD-FG group showed significantly higher compressive strength.ConclusionIncorporating 2 wt% FG into BD increases ion release,hydroxyapatite formation,and mechanical properties without compromising cell viability and differentiation.Clinical relevanceThe addition of FG enhanced the bioactivity of Biodentine and improved its strength without showing cytotoxicity,making it a promising approach that needs further study.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12903-025-06947-7. View Publication

α/β-hydrolase domain-containing protein 11 (ABHD11) is a mitochondrial hydrolase that maintains the catalytic function of α-ketoglutarate dehydrogenase (α-KGDH),and its expression in CD4 + T-cells has been linked to remission status in rheumatoid arthritis (RA). However,the importance of ABHD11 in regulating T-cell metabolism and function is yet to be explored. Here,we show that pharmacological inhibition of ABHD11 dampens cytokine production by human and mouse T-cells. Mechanistically,the anti-inflammatory effects of ABHD11 inhibition are attributed to increased 24,25-epoxycholesterol (24,25-EC) biosynthesis and subsequent liver X receptor (LXR) activation,which arise from a compromised TCA cycle. The impaired cytokine profile established by ABHD11 inhibition is extended to two patient cohorts of autoimmunity. Importantly,using murine models of accelerated type 1 diabetes (T1D),we show that targeting ABHD11 suppresses cytokine production in antigen-specific T-cells and delays the onset of diabetes in vivo in female mice. Collectively,our work provides pre-clinical evidence that ABHD11 is an encouraging drug target in T-cell-mediated inflammation. α/β-hydrolase domain-containing protein 11 (ABHD11) is a mitochondrial hydrolase,and its expression in CD4 + T-cells has been linked to remission status in rheumatoid arthritis. Here the authors report that pharmacological inhibition of ABHD11 modulates T-cell effector function via increased 24,25-epoxycholesterol biosynthesis and subsequent liver X receptor activation. View Publication

Simple SummaryEndometrial cancer (EC) develops in an environment strongly modulated by the immune system,and changes in dendritic cells (mDCs and pDCs),and monocytes (MO) expressing PD-L1 and PD-L2 may influence disease progression. Our study demonstrated that EC patients have lower percentages of PD-L1-positive MO and pDCs,as well as PD-L2-positive MO and mDCs,compared to the control group. We also found changes in plasma levels of soluble forms of PD-1,PD-L1,and PD-L2,which correlated with tumor PD-L2 expression and clinical characteristics,such as BMI and disease FIGO stage. AbstractBackground: Endometrial cancer is one of the most common female genital cancers and poses a significant clinical problem due to its increasing incidence and variable prognosis depending on the stage of the disease. The development of EC is largely dependent on interactions with the immune system,including immune checkpoints (ICPs) such as PD-1,PD-L1,and PD-L2. The aim of our study was to evaluate the PD-1/PD-L1/PD-L2 pathway in EC and its clinical significance. Methods: The analysis was performed by flow cytometry on myeloid and plasmacytoid dendritic cells and monocytes (MO) in peripheral blood (PB). The concentration of sPD-1,sPD-L1,and sPD-L2 in plasma was determined by ELISA. Additionally,PD-L1 and PD-L2 gene expression levels in tumor tissue (TT) were assessed using real-time polymerase chain reaction (qPCR). The obtained results were correlated with clinical data of EC patients. Results: Patients with EC had lower percentages of PD-L1-positive MO and pDCs,as well as PD-L2-positive MO and mDCs,compared with the control group. We observed accumulation of sPD-1 and lower levels of sPD-L1 and sPD-L2 in EC patients compared to the control group,with sPD-L2 correlating with PD-L2 gene expression level in the TT. Conclusions: The study results indicate a difference in the distribution of mDCs,pDCs,and MO with PD-L1/PD-L2 expression in EC patients. Reduced percentages of MO and DCs expressing PD-L1 and PD-L2,altered concentrations of soluble forms of these IPCs,and correlations with gene expression in TT suggest that dysregulation of this pathway may influence disease progression. Furthermore,the relationships between immunological parameters and clinical features such as BMI and FIGO stages suggest the potential use of these factors as diagnostic and prognostic biomarkers and the possibility of incorporating them into future therapeutic strategies. However,further studies are necessary to validate this hypothesis. View Publication

Background/Objectives: β-Carotene oxygenase-2 (BCO2) is a mitochondrial carotenoid-cleaving enzyme expressed in multiple tissues,including the lungs. While BCO2 regulates carotenoid handling,its role in shaping pulmonary immune architecture and antiviral responses is unknown. We hypothesized that BCO2 deficiency reprograms epithelial–innate circuits and alters antiviral outcomes. Methods: BCO2-knockout (KO) and C57BL/6J wild-type (WT) mice underwent lung single-cell RNA sequencing (scRNA-seq),immunoblotting,and intranasal SARS-CoV-2 challenge to assess cell-type heterogeneity,pathway programs (by gene set variation analysis,GSVA),and antiviral responses. Results: scRNA-seq resolved 14 major lung cell populations with cell-type-specific pathway shifts. Compared with WT,BCO2 KO lungs showed increased conventional dendritic cells and natural killer (NK) cells,with reductions in macrophages,B cells,and endothelial cells. In KO alveolar type II cells,GSVA indicated a stress-adapted metabolic program. Ciliated epithelium exhibited vitamin-K-responsive and axoneme-remodeling signatures with attenuated glucocorticoid and very-low-density lipoprotein remodeling. Innate lymphoid type 2 cells favored fatty acid oxidation and chromatin dynamics with reduced mitochondrial activity. NK cells were biased toward constitutive chemokine/cytokine secretion and counter-inflammatory signaling. Immunoblotting confirmed the elevated level of interferon regulatory factor-3 protein in BCO2-KO lungs. Functionally,BCO2-KO mice had improved outcomes after intranasal SARS-CoV-2 exposure. Conclusions: Loss of BCO2 reconfigures the pulmonary immune landscape and enhances antiviral responsiveness in mice. These findings identify BCO2 as a nutrient-linked enzyme with immunomodulatory impact and highlight cell-state changes as candidate mechanisms for improved antiviral tolerance. View Publication

Alterations in gut microbiota composition have been implicated in various diseases,including cancer. Recent evidence suggests that intestinal microbiota may influence the efficacy of immunotherapy. In this study,we investigated the relationship between gut dysbiosis and NK cell exhaustion in Mexican patients with cervical cancer (CC),a connection not previously explored. This cross-sectional study included newly diagnosed CC patients,a separate cohort of post-radio-chemotherapy (RCT) patients,and healthy donors (HD). Fecal microbiota profiles were assessed using 16S rRNA sequencing,while peripheral NK cell immune checkpoint expression was analyzed by multiparametric flow cytometry. CC patients exhibited significant gut dysbiosis,marked by reduced α-diversity,enrichment of pro-inflammatory taxa (Escherichia-Shigella,Prevotella),depletion of short-chain fatty acid (SCFA)-producing bacteria (Ruminococcus,Christensenellaceae),and enrichment of microbial metabolic pathways related to inflammation,oxidative stress,nutrient limitation,and immune suppression. Dysbiosis was more pronounced in patients after RCT,with further enrichment of Phascolarctobacterium. In parallel,NK cells displayed a putative exhausted phenotype,with elevated expression and co-expression of PD-1,LAG-3,TIM-3,TIGIT,BTLA,and NKG2A. A dysbiosis score and an NK exhaustion score were developed,revealing a significant positive correlation between microbial imbalance and NK cell exhaustion. Machine learning analysis identified the Escherichia/Ruminococcus ratio and PD-1+CD56bright NK cells as predictive markers of CC. Moreover,both dysbiosis and NK cell exhaustion markers were significantly associated with reduced patient survival. This is the first study to demonstrate a link between gut microbiota alterations and NK cell exhaustion in CC. Our findings suggest that gut dysbiosis may contribute to impaired anti-tumor immunity. This study supports the rationale for microbiota-targeted interventions as adjunctive strategies in CC,although prospective validation is required. View Publication

Over the past decade,non-viral DNA template delivery has been used with engineered nucleases to target single-stranded DNA sequences in hematopoietic stem and progenitor cells. While effective for gene therapy,this method is limited to short DNA donor templates,restricting its applications to gene corrections. To expand its scope,we developed an editing process using kilobase-long circular single-stranded DNA donor templates and TALEN technology. Our results show that the CssDNA editing process achieves high gene insertion frequency in HSPCs. Compared to AAV-edited HSPCs,CssDNA-edited HSPCs show a higher propensity to engraft and maintain gene edits in a female NCG murine model. This positive outcome is partly due to higher levels of primitive edited HSPCs,a more quiescent metabolic state,and elevated expression of bone marrow niche adhesion markers. Our findings highlight the strong potential of CssDNA as a universal,scalable and efficient non-viral DNA template for gene therapy applications. Letort et al. demonstrate that Circular single-stranded DNA enables efficient and precise gene insertion in hematopoietic stem cells,outperforming other DNA delivery formats and supporting progress toward scalable next-generation gene therapies. View Publication

AbstractTissue engineering for cardiovascular implants has largely utilized primary human cells to generate human tissue-engineered matrices (hTEMs). However,due to donor-to-donor variability and limited passage numbers,a more robust alternative to primary cells would be beneficial. To overcome these limitations,we have defined a new differentiation protocol for human-induced pluripotent stem cells (hiPSCs) into isogeneic cardiac fibroblast-like cells (iCFs) using animal sera-free and chemically defined methods. Morphology,extracellular matrix (ECM) deposition,and global transcriptomics revealed similarity between iCFs and primary human cardiac fibroblasts. Additionally,by overexpressing specific ECM and ECM-related proteins through gene-editing approaches,the ECM composition can be modulated as a building block to create “designer” next-generation hTEMs. Proteomics of gene-edited iCF-derived hTEMs demonstrated an increase in proteins involved in collagen and elastic fiber assembly. Furthermore,analysis of gene-edited iCF-derived hTEM mechanical functionality through biaxial mechanical testing exhibited increased collagen function,attributed to increased crosslinking and maturation. In sum,we have combined hiPSC technology with genome engineering to lay the foundation for next-generation tissue engineering applications by generating a novel cell source,gene-edited iCFs,that are able to modulate the composition as well as the functional mechanics of hTEMs. Graphical abstract Graphical Abstract Depicts the building blocks of next-generation human tissue-engineered matrices (hTEMs). Human-induced pluripotent stem cells (hiPSCs) as the starting cell source were gene edited,using the TALEN-based genome engineering approach,to induce a stable upregulation of genes involved in the biosynthesis and regulation of extracellular matrix (ECM) proteins (ie,lysyl oxidase [LOX],lysyl oxidase like 1 [LOXL1],elastin [ELN],and fibulin 5 [FBLN5]). These isogeneic hiPSCs were then differentiated into isogeneic cardiac fibroblast-like cells (iCFs) and used as building blocks to manufacture hTEMs for cardiovascular applications with modulated ECM composition and tissue mechanical properties. View Publication

The choroid plexus-cerebrospinal fluid (ChP-CSF) interface regulates a microenvironment supporting neural stem cell growth,strongly affected by hypoxia through ChP function. From human induced pluripotent stem cells (hiPSCs),here we established and validated in vitro ChP organoid secreting CSF-like fluid (iCSF) and exposed them to low oxygen atmosphere for 24 h. Transcriptomic indicated major data on morphological and functional alterations in the ChP cells and shotgun proteomics revealed significant changes in proteins involved in energy metabolism and mitochondrial function. We found that H2AZ and ITM2B,involved in neurogenesis and neurite growth,were the key proteins downregulated in hypoxic iCSF and ChP organoids,respectively. Positive correlation analysis between hypoxia-induced mRNA expression of the neuronal progenitor biomarkers SOX2 and PAX6. Mature neuron MAP2 and H2AZ also confirmed impairment of neurogenesis. The results from this study suggest that ChP-CSF interface opens new opportunities to characterize hypoxic brain pathophysiology and discover novel biomarkers.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12987-025-00731-z. View Publication

AbstractINTRODUCTIONRecent studies have identified important species‐dependent differences in the response of microglia to β‐amyloid (Aβ) pathology. Yet,whether human microglia also interact differently with the pathognomonic combination of amyloid and tau pathologies that occur in Alzheimer's disease (AD) remains unclear.METHODSWe generated a xenotolerant mouse model of AD that develops both plaque and tangle pathologies,transplanted stem cell‐derived microglial progenitors and examined the interactions between human microglia and AD pathologies with scRNA sequencing,immunohistochemistry,and in vitro modeling.RESULTSThe combined amyloid and tau pathologies induced robust type‐I interferon and proinflammatory cytokine responses,as well as an increased adoption of a distinct “rod” morphology in human microglia. The rod morphology could be induced with type‐I interferon treatment in vitro.DISCUSSIONWe provide new insights into human microglial responses to combined AD pathologies and a novel platform to investigate and manipulate human microglia in vivo.Highlights Amyloid pathology promotes the rapid development of neurofibrillary tangles and neuronal loss in a novel chimeric model of AD.Combined Alzheimer's disease pathologies lead to an expansion of disease‐associated microglia (DAM) and exacerbate Interferon‐responsive and cytokine/chemokine‐enriched states in xenotransplanted human microglia.The combination of amyloid and tau promotes the development of a distinctive rod microglial phenotype that closely correlates with tau pathology and neurodegeneration.Rod morphology and transcriptional changes can be modeled in vitro by treatment of induced pluripotent stem cells (iPSC) ‐microglia with type‐I interferons. View Publication

BACKGROUND: Glioblastoma (GBM),the most aggressive primary brain tumor,has a dismal prognosis largely due to therapy-resistant stem-like cells that drive recurrence. While N6-methyladenosine modifications in GBM are well-studied,the role of 5-methylcytosine (m5C) modifications specifically in circular RNAs (circRNAs) remains poorly understood. METHODS: Bioinformatic analysis,qRT-PCR and Western blot assays were used to investigate the expression of NSUN7,circNTRK2,YBX3,STK31,IKZF1 in GBM tissues and cell lines. m5C dot blot assays,m5C-bisulfite sequence assays,Sanger’s sequencing,RNA pull down and RIP assays,in vitro kinase assays,chromatin immunoprecipitation and luciferase reporter gene assays are used to clarify the interaction between factors above. Colony formation assays,sphere formation assays,and stemness marker (OCT4,DCLK1) analysis were utilized to assess the impact of the factors above on GBM stemness. Subcutaneous heterotopic and orthotopic xenograft are utilized to demonstrate the function of YBX3/circNTRK2/STK31/IKZF1 axis in GBM in vivo. RESULTS: NSUN7 and YBX3 are both upregulated in GBM tissues and cell lines. NSUN7 catalyzes m5C modification of circNTRK2. YBX3 decreasing circNTRK2 stability and function via binding m5C of circNTRK2. CircNTRK2 binds to and activates STK31,leading to phosphorylation of IKZF1 at S63,which decreases the O-GlcNAc levels and transcriptional activity of IKZF1,thereby promoting the stem cell characteristics of GBM cells. CONCLUSIONS: This study unveils that NSUN7-mediated m5C modification of circNTRK2 fuels GBM stemness via circNTRK2-dependent STK31 activation,thus identifying potential biomarkers for targeted molecular therapy of GBM and providing novel therapeutic targets for GBM treatment. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12967-025-07484-1. View Publication

Background: Cryptococcus-associated immune reconstitution inflammatory syndrome (C-IRIS) is a life-threatening complication of immune recovery,often triggered by antiretroviral therapy and characterized by Th1-skewed CD4+ T cell hyperactivation,neuroinflammation,and pulmonary dysfunction. Methods: Using a validated murine model of unmasking C-IRIS,we assessed the therapeutic potential of star-shaped glatiramer acetate (sGA),a structurally enhanced derivative of the FDA-approved immunomodulator glatiramer acetate (GA). sGA was administered intraperitoneally on days 1 and 3 post-CD4+ T cell reconstitution. Results: sGA significantly ameliorated C-IRIS-associated respiratory dysfunction,including increasing breaths per minute by ~35% and improved minute volume,total respiratory cycle time,expiration time,and inspiration time. Survival rate grew to 75% on day 14 for sGA-treated C-IRIS mice. In both the lung and the brain,sGA reduced total CD4+ T cells and selectively diminished Th1 cells by 50–60% and Th17 cells by 40–50%. Activated microglia decreased by 45% within the brain,indicating attenuated innate immune activation. Golgi-Cox analysis revealed region-specific neuroprotection: neuronal loss in the prefrontal cortex,lateral hypothalamus,and periaqueductal gray was rescued by 25–40%,whereas hippocampal neurons were relatively preserved,and basolateral amygdala neurons showed no significant recovery. Conclusion: Collectively,our findings suggest that sGA exerts neuroprotection in C-IRIS by limiting peripheral CD4+ T cell effector activity and suppressing CNS-resident immune activation. This study supports the use of sGA as a promising preclinical therapeutic candidate for C-IRIS and other Th1-mediated neuroinflammatory conditions. View Publication