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Oncogenic alterations in fibroblast growth factor receptor (FGFR)-family proteins occur across cancers,including pediatric gliomas. Our genomic analysis of 11,635 gliomas across ages finds that 5.3% of all gliomas harbor FGFR alterations,with an incidence of almost 9% in pediatric gliomas. Alterations in FGFR proteins are differentially enriched by age,tumor grade,and histology,with FGFR1 alterations associated with glioneuronal histologies. Leveraging isogenic systems,we confirm FGFR1 alterations to induce downstream Mitogen Activated Protein Kinase (MAPK) and mTOR signaling pathways,drive gliomagenesis,activate neuronal transcriptional programs and exhibit sensitivity to MAPK pathway and pan-FGFR inhibitors. Finally,we perform a retrospective analysis of clinical responses in children diagnosed with FGFR-altered gliomas and find that treatment with currently available inhibitors is largely associated with stability of disease. This study provides key insights into the biology of FGFR1-altered gliomas,therapeutic strategies to target them and associated challenges that still need to be overcome. A subset of pediatric gliomas harbour alterations in fibroblast growth factor receptor (FGFR)-family proteins. Here,the authors characterise the genomic landscape of 11,635 gliomas across ages and use isogenic model systems to explore the underlying biology of FGFR1-altered gliomas and potential therapeutic vulnerabilities. View Publication

AbstractMacrophages engulf apoptotic bodies and cellular debris as part of homeostasis,but they can also phagocytose live cells,such as aged red blood cells. Pharmacologic reprogramming with the SMAC mimetic LCL161 in combination with T-cell-derived cytokines can induce macrophages to phagocytose live cancer cells in mouse models. Here we extend these findings to encompass a wide range of monovalent and bivalent SMAC mimetic compounds,demonstrating that live cell phagocytosis is a class effect of these agents. We demonstrate robust phagocytosis of live pancreatic and breast cancer cells by primary human macrophages across a range of healthy donors. Unlike mouse macrophages,where a combination of SMAC mimetics with lymphotoxin enhanced phagocytosis,human macrophages were more efficiently polarized to phagocytose live cells by the combination of SMAC mimetics and IFNg. We profiled phagocytic macrophages by transcriptional and proteomic methodologies,uncovering a positive feedback loop of autocrine TNFa production. Graphical Abstract Graphical Abstract View Publication

BackgroundExtracellular Vesicles (EVs) derived from mesenchymal stem cells (MSCs) have gained recognition as promising therapeutic and drug delivery agents in regenerative medicine. However,their clinical application is limited by donor variability,low scalability,and inconsistent therapeutic quality. To overcome these challenges,a robust and standardized production platform is urgently needed.MethodsWe developed a scalable biomanufacturing strategy by generating and expanding MSCs from extended pluripotent stem cells (EPSC) using a suspension bioreactor culture system. A fixed-bed bioreactor was integrated for automated,continuous expansion of iMSCs and downstream EV harvesting. EVs were isolated through a streamlined protocol and characterized for size,morphology,surface markers,and bioactivity. Therapeutic efficacy was assessed in a bleomycin-induced pulmonary fibrosis mouse model.ResultsiMSC-derived EVs (iMSC-EVs) exhibited comparable characteristics to primary MSC-EVs,including a size distribution of 70–80 nm,cup-shaped morphology,and expression of canonical EV markers (CD63,CD81,TSG101). iMSCs were expanded for up to 20 days in 3D culture,yielding > 5 × 10⁸ cells per batch using a suspension bioreactor culture system and producing ~ 1.2 × 10¹³ EV particles/day in a fixed-bed bioreactor. In vivo,iMSC-EVs significantly reduced Ashcroft fibrosis scores and bronchoalveolar lavage fluid protein levels in bleomycin-injured lungs,with therapeutic efficacy comparable to primary MSC-EVs.ConclusionsThis study establishes a scalable and standardized platform for producing high-quality iMSC-EVs using bioreactor-based systems. Our approach addresses key limitations in traditional EV production and sets the stage for AI-integrated,fully automated,GMP-compliant manufacturing of therapeutic EVs suitable for clinical translation.Supplementary InformationThe online version contains supplementary material available at 10.1186/s13287-025-04507-y. View Publication

The telomerase enzyme is essential for telomere maintenance. Pathogenic variants in telomere-associated genes have been associated with critical telomere shortening,resulting in telomere biology disorders (TBD) such as bone marrow failure,idiopathic pulmonary fibrosis,and dyskeratosis congenita. The TBDs are clinically heterogeneous and families with TBD often experience an earlier onset and increased symptom severity for each generation. Consensus guidelines have identified certain genetic variants as pathogenic or likely pathogenic,but many are classified as variants of uncertain significance (VUS) in the absence of additional supporting evidence. The pathogenicity of a VUS in genes encoding the telomerase complex could be evaluated by in vitro telomerase activity (TA) measurement. We have developed a functional TA assay in patient-derived T-cells based on the Telomeric Repeat Amplification Protocol (TRAP) combined with qPCR. TA was significantly lower in six TBD patients with a TERT or TERC variant compared to controls (0.11 versus 0.54,p < 0.001). Four patients had a TA of more than three standard deviations below the mean of controls,strongly supporting pathogenicity of the variants. In summary,functional analysis of TA in patient-derived cells could support pathogenic evaluation in clinical diagnostics and reduce the number of reported VUS for TBD patients. View Publication

Given the local and systemic adverse reactions associated with whole-cell pertussis vaccines combined with diphtheria and tetanus toxoids (DTP),acellular pertussis vaccines combined with the same toxoids (DTaP) were developed in the 1990s. In comparison to DTP,DTaP vaccines demonstrated reduced reactogenicity and equivalent or improved immunogenicity and efficacy. However,there has been a resurgence of pertussis disease,particularly in DTaP-vaccinated children,suggesting that immunity wanes more quickly with DTaP vaccination. To elucidate the differences in immune responses to DTP and DTaP vaccines,we employed a systems biology-based strategy to compare global changes in gene expression following primary vaccination with either DTP or DTaP. We used RNA-Seq and ribosome profiling (RP) to identify transcriptional and translational signatures,respectively,in peripheral blood mononuclear cells (PBMCs) collected from 50 infant recipients of DTP or DTaP at two time-points (baseline (pre-vaccination at Day 1) and either Day 2 or 8 post-vaccination). We also used standard serologic methods to assess immunogenicity,and correlated these results with transcriptional and translational signatures. Here,we provide a detailed description of the rationale,experimental design,methodology,and enrollment procedures used. Given the technical complexity of our approach,our objective is to fill knowledge gaps,describe key quality metrics,and support future publications. In brief,we recovered 4-12 million PBMCs (average 8.9 million) with 99% viability per 2.5 mL blood sample,enabling excellent nucleic acid recovery yields for the preparation of high-quality sequencing libraries. In turn,these generated RNA-Seq and RP datasets with sufficient genome coverage breadth and depth to enable differential gene expression analyses,demonstrating the validity of this approach to study pertussis vaccine immunology specifically,and its utility to characterize mechanisms of the human immune response to vaccination generally. View Publication

Despite robust antibody responses,immunity induced by acellular pertussis vaccine (DTaP) wanes over time and risk of pertussis seems to be lower in children who receive whole-cell vaccine (DTP) as their first dose. To interrogate the early immunologic response to pertussis vaccine,we enrolled 56 healthy infants who received either DTP or DTaP at 2-,4-,6-,and 18-months of age. RNA-sequencing and ribosome profiling of PBMC were performed prior to vaccination (Day 1) and on either Day 2 or Day 8. Pathway enrichment analysis on Days 2 and 8 showed enrichment of TLR-signaling and FcϒR-mediated phagocytosis among DTP recipients. DTP also led to increases in IRAK-4 and IL-1ß. After booster vaccination,a higher frequency of PT-specific B-cells was observed in DTP- vs. DTaP recipients. These data provide insights into the early immunologic responses to pertussis vaccine and may guide next-generation pertussis vaccine development. View Publication

Porcine epidemic diarrhoea virus (PEDV) is the most widespread porcine coronavirus worldwide,causing high mortality and a high incidence rate among piglets. The molecular mechanisms by which PEDV regulates epithelial cell function and differentiation,as well as its disruption of the intestinal mucosal barrier,are not yet fully understood. This study reveals that PEDV infection reduces the number of goblet cells and impairs the intestinal barrier integrity in newborn piglets. Regarding the pathways involved in the differentiation of intestinal stem cells (ISCs),PEDV infection concurrently activates the Notch and MAPK pathways while suppressing the Wnt/β-catenin pathway in the intestines of piglets. Furthermore,in vitro experiments using intestinal monolayer organoid models showed that PEDV infection hinders goblet cell differentiation by activating the Notch signalling pathway. Additionally,the PEDV-encoded ORF3 protein,which is crucial for activating the Notch pathway,inhibits goblet cell differentiation in PEDV-infected intestinal monolayer organoids. This study offers new insights into the mechanisms underlying intestinal mucosal barrier dysfunction induced by PEDV infection.Supplementary InformationThe online version contains supplementary material available at 10.1186/s13567-025-01599-5. View Publication

AbstractPancreatic ductal adenocarcinoma (PDAC),one of the most aggressive forms of pancreatic cancer,is associated with poor survival outcomes and currently ranks as the third leading cause of cancer-related death in the United States. Despite its clinical significance,the mechanisms of PDAC development and progression remain,in part,poorly understood. In this study,we provide evidence of a novel role of sorting nexin 10 (SNX10),a member of the sorting nexin family,in the regulation of KRAS-induced pancreatic carcinogenesis. We demonstrate that SNX10 is downregulated in PDAC,especially in advanced cases. Furthermore,mutational analysis revealed SNX10 genetic alterations in PDAC cases. Functional studies demonstrated that SNX10 overexpression in human PDAC cells inhibited cell proliferation and colony formation. Moreover,SNX10 overexpression induced G1-phase cell-cycle arrest and decreased KRAS signaling activity. Using a novel Snx10 knockout mouse crossed with a Kras-driven PDAC model,we observed reduced survival,increased tumor cell proliferation,enhanced aggression,and heightened inflammation. Collectively,these findings highlight SNX10 as a tumor suppressor candidate in PDAC and underscore its promise as a foundation for new therapeutic approaches.Significance:SNX10 plays a crucial role in reducing pancreatic tumorigenesis. This discovery offers valuable insights into PDAC’s biology and the development of new effective treatments. View Publication

Epilepsy is a central nervous system disorder causing uncontrollable seizures. One-third of patients do not respond to current medications,necessitating new treatments. This study targeted epileptogenesis,the process leading to chronic epilepsy,using human mesenchymal stem cells (MSCs) in a rodent model. MSC transplantation can positively affect neurodegenerative diseases by modifying inflammation. Additionally,glial cell line-derived neurotrophic factor (GDNF) may counteract seizures and tissue damage. We transplanted naïve immortalized human adipose-derived MSCs (Ctrl-MSCs) or GDNF-releasing MSCs (GDNF-MSCs,releasing 588.67 ± 20.14 pg/ml/24 h GDNF) into rat hippocampi after kainic acid-induced status epilepticus. Seizure progression was monitored for 5 weeks using video-EEG,behavioral assessments,and histological analysis. Both cell types influenced epileptogenesis. GDNF-MSCs delayed early-stage seizures,while Ctrl-MSCs reduced seizure frequency in later stages. Differences emerged in seizure development and cumulative seizure count,with Ctrl-MSCs showing significant seizure-attenuating effects. Behavioral differences were also noted: Ctrl-MSCs improved short-term memory and reduced anxiety,whereas GDNF-MSCs primarily reduced anxiety without significantly improving memory. This study highlights the therapeutic potential of MSCs,with or without GDNF,in modulating epileptogenesis,offering promising avenues for future clinical treatments.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00018-025-05853-z. View Publication

AbstractCentral adiposity and poor cardiorespiratory fitness are modifiable risk factors for various diseases. This study investigated their impact on CD4+ differentiated T regulatory (Treg) cell responses. Thirty‐eight young adults were classified into high cardiorespiratory fitness/low visceral adipose tissue (High V̇O2–Low VAT,n = 20) and low cardiorespiratory fitness/high VAT (Low V̇O2–High VAT,n = 18). Body composition was assessed using DXA and ultrasound,while cardiorespiratory fitness and physical activity were measured via treadmill testing and accelerometry. CD4+ cells were cultured in Treg differentiation medium with 2 ng/mL TGF‐β,with or without 100 nM rapamycin or 50 nM Torin‐1,for 96 h. Differentiated Treg from Low V̇O2–High VAT participants exhibited no significant changes in IL‐10 or IL‐6 production with rapamycin or Torin‐1. Conversely,differentiated Treg from High V̇O2–Low VAT participants showed significantly lower IL‐10 production with rapamycin (p < 0.001,adjusted p < 0.001) and Torin‐1 (p < 0.001,adjusted p < 0.001). These findings indicate that low cardiorespiratory fitness and high VAT contribute to an altered inflammatory response,influencing peripheral blood mononuclear cell immunophenotypes and exhaustion markers. Furthermore,mTORC1 and mTORC2 inhibition modulate cytokine production,emphasizing the role of metabolic status in immune regulation. High cardiorespiratory fitness and low visceral adiposity modulate CD4+ T regulatory cells. Fit participants shows be more responsible to mTORC1/2 inhibition via rapamycin or Torin‐1indicating a fitness‐ and adiposity‐dependent immunometabolic regulation. View Publication

The zinc containing matrix metalloproteinase enzyme regulates a diverse array of biological processes in health and disease,including ADAM17 (a disintegrin and metalloproteinase domain 17) enzyme. Due to its large substrate profile,ADAM17 is known to regulate diverse pathways of inflammation and adaptive immunity. However,the role of ADAM17 in modulating the pathogenesis of type 2 allergic asthma is largely unknown. To determine the in vivo contribution of ADAM17 in house dust mite (HDM)-induced airway inflammation and adaptive immune response,we assessed the deletion of ADAM17 in mice conventional dendritic cells (ΔDC) and employed a complementary chemical biology approach using small-molecule novel ADAM17 inhibitor (2155-17). DC-specific ADAM17 ablation (ΔDC) suppressed type 2/ eosinophilic polarized HDM allergic responses and is protected from developing AHR. DC isolated from ΔDC mice showed a reduced state of metabolic activity,immune priming function and suppressed allergen-specific type 2 cell polarizations. Intranasal administration of 2155-17 protected WT mice against type2/ eosinophilic polarized HDM allergic responses. These concurrent results from two independent approaches identify a novel role for ADAM17 as an upstream site in airway inflammation. Furthermore,targeting ADAM17 with a selective small-molecule inhibitor might be harnessed as a potential drug target for type 2-high allergic asthma. View Publication

The mechanisms through which mutations in splicing factor genes drive clonal hematopoiesis (CH) and myeloid malignancies,and their close association with advanced age,remain poorly understood. Here we show that telomere maintenance plays an important role in this phenomenon. First,by studying 454,098 UK Biobank participants,we find that,unlike most CH subtypes,splicing-factor-mutant CH is more common in those with shorter genetically predicted telomeres,as is CH with mutations in PPM1D and the TERT gene promoter. We go on to show that telomere attrition becomes an instrument for clonal selection in advanced age,with splicing factor mutations ‘rescuing’ HSCs from critical telomere shortening. Our findings expose the lifelong influence of telomere maintenance on hematopoiesis and identify a potential shared mechanism through which different splicing factor mutations drive leukemogenesis. Understanding the mechanistic basis of these observations can open new therapeutic avenues against splicing-factor-mutant CH and hematological or other cancers. This study explores the relationship between telomere length and clonal hematopoiesis. Splicing factor and PPM1D gene mutations are more frequent in people with genetically predicted shorter telomere lengths,suggesting that these mutations protect against the consequences of telomere attrition. View Publication