参考文献

Mural cells are central to vascular integrity and function. In this study,we demonstrate the innovative use of the transcription factor NKX3.1 to guide the differentiation of human induced pluripotent stem cells into mural progenitor cells (iMPCs). By transiently activating NKX3.1 in mesodermal intermediates,we developed a method that diverges from traditional growth factor-based differentiation techniques. This approach efficiently generates a robust iMPC population capable of maturing into diverse functional mural cell subtypes,including smooth muscle cells and pericytes. These iMPCs exhibit key mural cell functionalities such as contractility,deposition of extracellular matrix,and the ability to support endothelial cell-mediated vascular network formation in vivo. Our study not only underscores the fate-determining significance of NKX3.1 in mural cell differentiation but also highlights the therapeutic potential of these iMPCs. We envision these insights could pave the way for a broader use of iMPCs in vascular biology and regenerative medicine. Mural progenitor cells are crucial for vascular stability. Here,the authors generate these cells from human pluripotent stem cells using NKX3.1 and show that they can mature into various mural cell types and contribute to blood vessel formation. View Publication

The generation of retinal models from human induced pluripotent stem cells holds significant potential for advancing our understanding of retinal development,neurodegeneration,and the in vitro modeling of neurodegenerative disorders. The retina,as an accessible part of the central nervous system,offers a unique window into these processes,making it invaluable for both study and early diagnosis. This study investigates the impact of the Frontotemporal Dementia-linked IVS 10?+?16 MAPT mutation on retinal development and function using 2D and 3D retinal models derived from human induced pluripotent stem cells. Our findings reveal that the MAPT mutation leads to delayed retinal cell differentiation and maturation,with tau-mutant disease models exhibiting sustained higher expression of retinal progenitor cell markers and a reduced presence of post-mitotic neurons. Both 2D and 3D tau-mutant retinal models demonstrated an imbalance in tau isoforms,favoring 4R tau,along with increased tau phosphorylation,altered neurite morphology,and impaired cytoskeletal maturation. These changes are associated with impaired synaptic development,reduced neuronal connectivity,and enhanced cellular stress responses,including the increased formation of stress granules,markers of apoptosis and autophagy,and the presence of intracellular toxic tau aggregates. This study highlights the value of retinal models derived from human induced pluripotent stem cells in exploring the mechanisms underlying retinal pathology associated with tau mutations. These models offer essential insights into the development of therapeutic strategies for neurodegenerative diseases characterized by tau aggregation.Supplementary InformationThe online version contains supplementary material available at 10.1186/s40478-024-01920-x. View Publication

In Finland,the frequency of isolated cleft palate (CP) is higher than that of isolated cleft lip with or without cleft palate (CL/P). This trend contrasts to that in other European countries but its genetic underpinnings are unknown. We conducted a genome-wide association study in the Finnish population and identified rs570516915,a single nucleotide polymorphism highly enriched in Finns,as strongly associated with CP (P?=?5.25 × 10?34,OR?=?8.65,95% CI 6.11–12.25),but not with CL/P (P?=?7.2 × 10?5),with genome-wide significance. The risk allele frequency of rs570516915 parallels the regional variation of CP prevalence in Finland,and the association was replicated in independent cohorts of CP cases from Finland (P?=?8.82 × 10?28) and Estonia (P?=?1.25 × 10?5). The risk allele of rs570516915 alters a conserved binding site for the transcription factor IRF6 within an enhancer (MCS-9.7) upstream of the IRF6 gene and diminishes the enhancer activity. Oral epithelial cells derived from CRISPR-Cas9 edited induced pluripotent stem cells demonstrate that the CP-associated allele of rs570516915 concomitantly decreases the binding of IRF6 and the expression level of IRF6,suggesting impaired IRF6 autoregulation as a molecular mechanism underlying the risk for CP. Here,the authors perform a genome-wide study and identify a genetic variant enriched in the Finnish population that is strongly associated with isolated cleft palate. This finding suggests a genetic basis for the high prevalence of cleft palate in Finland. View Publication

Increasing shreds of evidence suggest that neurogenic-to-gliogenic shift may be critical to the abnormal neurodevelopment observed in individuals with Down syndrome (DS). REST,the Repressor Element-1 Silencing Transcription factor,regulates the differentiation and development of neural cells. Downregulation of REST may lead to defects in post-differentiation neuronal morphology in the brain of the DS fetal. This study aims to elucidate the role of REST in DS-derived NPCs using bioinformatics analyses and laboratory validations. We identified and validated vital REST-targeted DEGs: CD44,TGFB1,FN1,ITGB1,and COL1A1. Interestingly,these genes are involved in neurogenesis and gliogenesis in DS-derived NPCs. Furthermore,we identified nuclear REST loss and the neuroblast marker,DCX,was downregulated in DS human trisomic induced pluripotent stem cells (hiPSCs)-derived NPCs,whereas the glioblast marker,NFIA,was upregulated. Our findings indicate that the loss of REST is critical in the neurogenic-to-gliogenic shift observed in DS-derived NPCs. REST and its target genes may collectively regulate the NPC phenotype.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-87314-y. View Publication

Peroxisomes are eukaryotic organelles that are essential for multiple metabolic pathways,including fatty acid oxidation,degradation of amino acids,and biosynthesis of ether lipids. Consequently,peroxisome dysfunction leads to pediatric-onset neurodegenerative conditions,including Peroxisome Biogenesis Disorders (PBD). Due to the dynamic,tissue-specific,and context-dependent nature of their biogenesis and function,live cell imaging of peroxisomes is essential for studying peroxisome regulation,as well as for the diagnosis of PBD-linked abnormalities. However,the peroxisomal imaging toolkit is lacking in many respects,with no reporters for substrate import,nor cell-permeable probes that could stain dysfunctional peroxisomes. Here we report that the BODIPY-C12 fluorescent fatty acid probe stains functional and dysfunctional peroxisomes in live mammalian cells. We then go on to improve BODIPY-C12,generating peroxisome-specific reagents,PeroxiSPY650 and PeroxiSPY555. These probes combine high peroxisome specificity,bright fluorescence in the red and far-red spectrum,and fast non-cytotoxic staining,making them ideal tools for live cell,whole organism,or tissue imaging of peroxisomes. Finally,we demonstrate that PeroxiSPY enables diagnosis of peroxisome abnormalities in the PBD CRISPR/Cas9 cell models and patient-derived cell lines. The array of tools to image peroxisome regulation is still limited. Here,the authors develop improved fatty acid-based probes with high peroxisome specificity and bright fluorescence in the red/far-red spectrum,which makes them ideal to study peroxisomes in live cells and whole organisms. View Publication

Somatic cell reprogramming into human induced pluripotent stem cells entails significant intracellular changes,including modifications in mitochondrial metabolism and a decrease in mitochondrial DNA copy number. However,the mechanisms underlying this decrease in mitochondrial DNA copy number during reprogramming remain unclear. Here we aimed to elucidate these underlying mechanisms. Through a meta-analysis of several RNA sequencing datasets,we identified genes responsible for the decrease in mitochondrial DNA. We investigated the functions of these identified genes and assessed their regulatory mechanisms. In particular,the expression of the thymidine kinase 2 gene (TK2),located in the mitochondria and required for mitochondrial DNA synthesis,is decreased in human pluripotent stem cells as compared with its expression in somatic cells. TK2 was significantly downregulated during reprogramming and markedly upregulated during differentiation. Collectively,this decrease in TK2 levels induces a decrease in mitochondrial DNA copy number and contributes to shaping the metabolic characteristics of human pluripotent stem cells. However,contrary to our expectations,treatment with a TK2 inhibitor impaired somatic cell reprogramming. These results suggest that decreased TK2 expression may result from metabolic conversion during somatic cell reprogramming. Mitochondrial DNA loss linked to stem cell reprogrammingInduced pluripotent stem (iPS) cells are special cells created by reprogramming regular body cells. Researchers explored how these cells change their energy production methods during reprogramming. The study focused on a protein called thymidine kinase 2 (TK2),which is important for maintaining mitochondrial DNA (mtDNA). Mitochondria are the cell’s powerhouses,and their DNA is crucial for energy production. Researchers used human cell lines to study how TK2 affects mtDNA during reprogramming. They found that,as cells become iPS cells,TK2 levels drop,leading to reduced mtDNA and a shift in energy production from oxidative phosphorylation to glycolysis. Results suggest that reducing TK2 and mtDNA is key for cells to gain pluripotency. This shift helps support the rapid growth and development of iPS cells. Understanding this process could improve stem cell therapies and regenerative medicine in the future.This summary was initially drafted using artificial intelligence,then revised and fact-checked by the author. View Publication

The angiotensin-converting enzyme 2 (ACE2) is a primary cell surface viral binding receptor for SARS-CoV-2,so finding new regulatory molecules to modulate ACE2 expression levels is a promising strategy against COVID-19. In the current study,we utilized islet organoids derived from human embryonic stem cells (hESCs),animal models and COVID-19 patients to discover that fibroblast growth factor 7 (FGF7) enhances ACE2 expression within the islets,facilitating SARS-CoV-2 infection and resulting in impaired insulin secretion. Using hESC-derived islet organoids,we demonstrated that FGF7 interacts with FGF receptor 2 (FGFR2) and FGFR1 to upregulate ACE2 expression predominantly in ? cells. This upregulation increases both insulin secretion and susceptibility of ? cells to SARS-CoV-2 infection. Inhibiting FGFR counteracts the FGF7-induced ACE2 upregulation,subsequently reducing viral infection and replication in the islets. Furthermore,retrospective clinical data revealed that diabetic patients with severe COVID-19 symptoms exhibited elevated serum FGF7 levels compared to those with mild symptoms. Finally,animal experiments indicated that SARS-CoV-2 infection increased pancreatic FGF7 levels,resulting in a reduction of insulin concentrations in situ. Taken together,our research offers a potential regulatory strategy for ACE2 by controlling FGF7,thereby protecting islets from SARS-CoV-2 infection and preventing the progression of diabetes in the context of COVID-19. View Publication

Advancements in vaccination and sanitation have significantly reduced the prevalence and burden of infectious diseases; however,these benefits have coincided with a marked rise in autoimmune and allergic disorders. Recent studies have investigated these linked trends through the lens of host-microbiome alterations,proposing these shifts as a potential explanatory mechanism. Previously,we demonstrated that vancomycin-induced depletion of short-chain fatty acid (SCFA)-producing bacteria results in hyperactivation of ILC2s and exacerbated allergic responses. Here we investigate the effects of low-dose streptomycin on innate and adaptive immune cell populations and their activation states. Although streptomycin-treated mice exhibit normal allergic responses,they display heightened susceptibility to Th1/Th17-mediated disease,specifically hypersensitivity pneumonitis (HP). This is characterized by a two-fold increase in ILC3s and Th17 cells in the lungs,alongside activation of antigen-presenting cells (APCs) at steady state-an effect that is further amplified upon exposure to HP-inducing agents. Shotgun metagenomic analysis revealed that streptomycin-induced dysbiosis reduces microbial diversity,depletes bile acid-metabolizing bacteria,and enriches for metabolic pathways involved in branched-chain amino acid biosynthesis,including leucine-a known activator of mTORC1. Strikingly,administration of the secondary bile acid metabolite isolithocholic acid (an inverse agonist of RORγt),or an IL-23 neutralizing antibody,reverses the enhanced susceptibility to HP. Inhibition of mTORC1 significantly reduced Th17/ILC3 responses and histopathology. Our findings underscore microbial equilibrium as a key determinant of susceptibility to HP and uncover a positive feedback loop between IL and 23-producing APCs and ILC3/Th17 cells that mechanistically links dysbiosis to sustained type 3 inflammation,and we identify a simple,actionable means of intervention. View Publication

The association between type 2 diabetes (T2D) pathogenesis and immune-mediated tissue damage and insulin resistance suggests that T2D patients might benefit from the suppression of pathogenic inflammation. Foxp3+ Treg cells are crucial suppressors of inflammation,but the differentiation of Foxp3+ Treg cells is not static and is subject to conversion into IL-17-producing Th17-like cells upon receiving external signals. In this study,we examined the production of IL-17 by Treg cells. Compared to non-T2D controls,T2D patients presented significantly higher levels of IL-17-expressing cells in both Foxp3- CD4 T cells and Foxp3+ Treg cells. The frequencies of IL-17-nonexpressing Foxp3+ Treg cells,on the other hand,were not changed. Interestingly,IL-17-expressing Foxp3+ Treg cells were mutually exclusive from IL-10-expressing and TGF-$\beta$-expressing Foxp3+ Treg cells,suggesting that multiple subpopulations exist within the Foxp3+ Treg cells from T2D patients. In T2D patients,the frequencies of IL-17-expressing Foxp3+ Treg cells were positively correlated with the body mass index (BMI) and the HbA1c levels of T2D patients. The frequencies of IL-10-expressing Treg cells,on the other hand,were inversely associated with the BMI of both non-T2D controls and T2D patients. In addition,the suppressive activity of Treg cells was significantly lower in T2D patients than in non-T2D controls. Together,our study uncovered a dysregulation in Foxp3+ Treg cells from T2D patients,characterized by high IL-17 expression and low suppression activity. View Publication

Osteoporotic osteoarthritis (OPOA) is a common bone disease mostly in the elderly,but the relationship between Osteoporotic (OP) and osteoarthritis (OA) is complex. It has been shown that knee loading can mitigate OA symptoms. However,its effects on OPOA remain unclear. In this study,we characterized pathological linkage of OP to OA,and evaluated the effect of knee loading on OPOA. We employed two mouse models (OA and OPOA),and conducted histology,cytology,and molecular analyses. In the OA and OPOA groups,articular cartilage was degenerated and Osteoarthritis Research Society International score was increased. Subchondral bone underwent abnormal remodeling,the differentiation of bone marrow mesenchymal stem cells (BMSCs) to osteoblasts and chondrocytes was reduced,and migration and adhesion of pre-osteoclasts were enhanced. Compared to the OA group,the pathological changes of OA in the OPOA group were considerably aggravated. After knee loading,however,cartilage degradation was effectively prevented,and the abnormal remodeling of subchondral bone was significantly inhibited. The differentiation of BMSCs was also improved,and the expression of Wnt/$\beta$-catenin was elevated. Collectively,this study demonstrates that osteoporosis aggravates OA symptoms. Knee loading restores OPOA by regulating subchondral bone remodeling,and may provide an effective method for repairing OPOA. View Publication