Oligoasthenoteratozoospermia is an important factor affecting male fertility and has been found to be associated with genetic factors.However,there are stll a proportion of oligoasthenoteratozoospermia cases that cann...Oligoasthenoteratozoospermia is an important factor affecting male fertility and has been found to be associated with genetic factors.However,there are stll a proportion of oligoasthenoteratozoospermia cases that cannot be explained by known pathogenic genetic variants.Here,we perform genetic analyses and identify bi-allelic loss-of-function variants of MFSD6L from an oligoasthenoteratozoospermia-affected family.Mfsd6l knock-out male mice also present male subfertility with reduced sperm concentration,motility,and deformed acrosomes.Further mechanistic analyses reveal that MFsD6L,as an acrosome membrane protein,plays an important role in the formation of acrosome by interacting with the inner acrosomal membrane protein SPACA1.Moreover,poor embryonic development is consistently observed after intracytoplasmic sperm injection treatment using spermatozoa from the MFSD6L-deficient man and male mice.Collectively,our findings reveal that MFSD6L is required for the anchoring of sperm acrosome and head shaping.The deficiency of MFsD6L affects male fertility and causes oligoasthenoter-atozoospermia in humans and mice.展开更多
During spermiogenesis,haploid spermatids undergo dramatic morphological changes to form slender sperm flagella and cap-like acrosomes,which are required for successful fertilization.Severe deformities in flagella caus...During spermiogenesis,haploid spermatids undergo dramatic morphological changes to form slender sperm flagella and cap-like acrosomes,which are required for successful fertilization.Severe deformities in flagella cause a male infertility syndrome,multiple morphological abnormalities of the flagella(MMAF),while acrosomal hypoplasia in some cases leads to sub-optimal embryonic developmental potential.However,evidence regarding the occurrence of acrosomal hypoplasia in MMAF is limited.Here,we report the generation of base-edited mice knocked out for coiled-coil domain-containing 38(Ccdc38)via inducing a nonsense mutation and find that the males are infertile.The Ccdc38-KO sperm display acrosomal hypoplasia and typical MMAF phenotypes.We find that the acrosomal membrane is loosely anchored to the nucleus and fibrous sheaths are disorganized in Ccdc38-KO sperm.Further analyses reveal that Ccdc38 knockout causes a decreased level of TEKT3,a protein associated with acrosome biogenesis,in testes and an aberrant distribution of TEKT3 in sperm.We finally show that intracytoplasmic sperm injection overcomes Ccdc38-related infertility.Our study thus reveals a previously unknown role for CCDC38 in acrosome biogenesis and provides additional evidence for the occurrence of acrosomal hypoplasia in MMAF.展开更多
T-box transcription factor T(TBXT;T)is required for mesodermal formation and axial skeletal development.Although it has been extensively studied in various model organisms,human congenital vertebral malformations(CVMs...T-box transcription factor T(TBXT;T)is required for mesodermal formation and axial skeletal development.Although it has been extensively studied in various model organisms,human congenital vertebral malformations(CVMs)involving T are not well established.Here,we report a family with 15 CVM patients distributed across 4 generations.All affected individuals carry a heterozygous mutation,T c.596A>G(p.Q199R),which is not found in unaffected family members,indicating co-segregation of the genotype and phenotype.In vitro assays show that T p.Q199R increases the nucleocytoplasmic ratio and enhances its DNA-binding affinity,but reduces its transcriptional activity compared to the wild-type.To determine the pathogenicity of this mutation in vivo,we generated a Q199R knock-in mouse model that recapitulates the human CVM phenotype.Most heterozygous Q199R mice show subtle kinked or shortened tails,while homozygous mice exhibit tail filaments and severe vertebral deformities.Overall,we show that the Q199R mutation in T causes CVM in humans and mice,providing previously unreported evidence supporting the function of T in the genetic etiology of human CVM.展开更多
Sedentary behavior for two years during the coronavirus disease 2019(COVID-19)pandemic contributes to weight gain.Gut microbiota and blood metabolome are related to body mass index(BMI)and indicate individual metaboli...Sedentary behavior for two years during the coronavirus disease 2019(COVID-19)pandemic contributes to weight gain.Gut microbiota and blood metabolome are related to body mass index(BMI)and indicate individual metabolic changes.Surgery and exercise are effective weight-loss methods.The precise plasma metabolites and gut microbiota biomarkers involved and the underlying mechanisms are still largely unclear.To address this issue,we analyzed weight gain and weight loss cohorts to identify biomarkers associated with obesity.In the sedentary cohort,49 subjects were recruited in year 2019.After two years of sedentary behavior during the COVID-19 pandemic,the BMI of 24 subjects significantly increased(Weight gain group),while that of the remnant 25 subjects remained constant(Maintaining weight group).At baseline and two years post baseline,the gut microbiota and blood metabolome,as well as body composition and clinical indicators,were all collected.In weight loss studies,we analyze the plasma metabolome of the two cohorts,including individuals who underwent laparoscopic sleeve gastrectomy(LSG)surgery and exercise intervention.Weight gain through sedentary behavior contributed to the variation of the gut microbiota and plasma metabolites composition.Creatine,phenylalanine and tyrosine exhibited significant positive associations with BMI and fat mass.We further confirmed the association between BMI and plasma metabolites in two weight loss cohorts.By utilizing a linear regression model,we found that 10 metabolites including creatine were correlated with BMI in weight loss individuals.Based on receiver operating characteristic(ROC)curves,creatine exhibited a satisfactory classification performance in regard to predicting weight reduction(AUC_(LSG)=0.890,AUC_(Sports)=0.840).Moreover,some gut microbiota,including Bifidobacterium angulatum DSM 20098=JCM 7096 and Rothia dentocariosa M567I could affect BMI through the mediating factor of creatine.展开更多
Bromodomain and extra-terminal domain(BET)proteins,which function partly through MYC proto-oncogene(MYC),are critical epigenetic readers and emerging therapeutic targets in cancer.Whether and how BET inhibition simult...Bromodomain and extra-terminal domain(BET)proteins,which function partly through MYC proto-oncogene(MYC),are critical epigenetic readers and emerging therapeutic targets in cancer.Whether and how BET inhibition simultaneously induces metabolic remodeling in cancer cells remains unclear.Here we find that even transient BET inhibition by JQ-1 and other pan-BET inhibitors(pan-BETis)blunts liver cancer cell proliferation and tumor growth.BET inhibition decreases glycolytic gene expression but enhances mitochondrial glucose and glutamine oxidative metabolism revealed by metabolomics and isotope labeling analysis.Specifically,BET inhibition downregulates miR-30a to upregulate glutamate dehydrogenase 1(GDH1)independent of MYC,which producesα-ketoglutarate for mitochondrial oxidative phosphorylation(OXPHOS).Targeting GDH1 or OXPHOS is synthetic lethal to BET inhibi-tion,and combined BET and OXPHOS inhibition therapeutically prevents liver tumor growth in vitro and in vivo.Together,we uncover an important epigenetic-metabolic crosstalk whereby BET inhibition induces MYC-independent and GDH1-dependent glutamine metabolic remodeling that can be exploited for innovative combination therapy of liver cancer.展开更多
βcells are defined by the ability to produce and secret insulin.Recent studies have evaluated that human pancreaticβcells are heterogeneous and demonstrated the transcript alterations ofβcell subpopulation in diabe...βcells are defined by the ability to produce and secret insulin.Recent studies have evaluated that human pancreaticβcells are heterogeneous and demonstrated the transcript alterations ofβcell subpopulation in diabetes.Single-cell RNA sequence(scRNA-seq)analysis helps us to refine the cell types signatures and understand the role of theβcells during metabolic challenges and diseases.Here,we construct the pseudotime trajectory ofβcells from publicly available scRNA-seq data in health and type 2 diabetes(T2D)based on highly dispersed and highly expressed genes using Monocle2.We identified three major states including 1)Normal branch,2)Obesity-like branch and 3)T2D-like branch based on biomarker genes and genes that give rise to bifurcation in the trajectory.βcell function-maintain-related genes,insulin expression-related genes,and T2D-related genes enriched in three branches,respectively.Continuous pseudotime spectrum might suggest thatβcells transition among different states.The application of pseudotime analysis is conducted to clarify the different cell states,providing novel insights into the pathology ofβcells in T2D.展开更多
Cytosine base editing achieves C·G-to-T·A substitutions and can convert four codons(CAA/CAG/CGA/TGG)into STOP-codons(induction of STOP-codons,iSTOP)to knock out genes with reduced mosaicism.iSTOP enables dir...Cytosine base editing achieves C·G-to-T·A substitutions and can convert four codons(CAA/CAG/CGA/TGG)into STOP-codons(induction of STOP-codons,iSTOP)to knock out genes with reduced mosaicism.iSTOP enables direct phenotyping in founders’somatic cells,but it remains unknown whether this works in founders’germ cells so as to rapidly reveal novel genes for fertility.Here,we initially establish that iSTOP in mouse zygotes enables functional characterization of known genes in founders’germ cells:Cfap43-iSTOP male founders manifest expected sperm features resembling human“multiple morphological abnormalities of the flagella”syndrome(i.e.,MMAF-like features),while oocytes of Zp3-iSTOP female founders have no zona pellucida.We further illustrate iSTOP’s utility for dissecting the functions of unknown genes with Ccdc183,observing MMAF-like features and male infertility in Ccdc183-iSTOP founders,phenotypes concordant with those of Ccdc183-KO offspring.We ultimately establish that CCDC183 is essential for sperm morphogenesis through regulating the assembly of outer dynein arms and participating in the intra-flagellar transport.Our study demonstrates iSTOP as an efficient tool for direct reproductive disease modeling and phenotyping in germ cells of the founder generation,and rapidly reveals the essentiality of Ccdc183 in fertility,thus providing a time-saving approach for validating genetic defects(like nonsense mutations)for human infertility.展开更多
Dear Editor,Tetralogy of Fallot(TOF)is the most common complex congenital heart disease.Besides gene mutations and copy number variants,altered protein function induced by posttranscriptional or translational regulati...Dear Editor,Tetralogy of Fallot(TOF)is the most common complex congenital heart disease.Besides gene mutations and copy number variants,altered protein function induced by posttranscriptional or translational regulation also contributes to the onset of TOF.1 MiRNAs are short noncoding RNAs that bind to the 3’-UTR of target mRNAs to repress protein production.However,the causal link between miRNAs and TOF and the underlying mechanism has not been established.展开更多
Metabolically healthy obesity refers to obese individuals who do not develop metabolic disorders.These people store fat in subcutaneous adipose tissue(SAT)rather than in visceral adipose tissue(VAT).However,the molecu...Metabolically healthy obesity refers to obese individuals who do not develop metabolic disorders.These people store fat in subcutaneous adipose tissue(SAT)rather than in visceral adipose tissue(VAT).However,the molecules participating in this specific scenario remain elusive.Rab18,a lipid droplet(LD)-associated protein,mediates the contact between the endoplasmic reticulum(ER)and LDs to facilitate LD growth and maturation.In the present study,we show that the protein level of Rab18 is specifically upregulated in the SAT of obese people and mice.Rab18 adipocyte-specific knockout(Rab18 AKO)mice had a decreased volume ratio of SAT to VAT compared with wildtype mice.When subjected to high-fat diet(HFD),Rab18 AKO mice had increased ER stress and inflammation,reduced adiponectin,and decreased triacylglycerol(TAG)accumulation in SAT.In contrast,TAG accumulation in VAT,brown adipose tissue(BAT)or liver of Rab18AKO mice had a moderate increase without ER stress stimulation.Rab18 AKO mice developed insulin resistance and systematic inflammation.Rab18 AKO mice maintained body temperature in response to acute and chronic cold induction with a thermogenic SAT,similar to the counterpart mice.Furthermore,Rab18-deficient 3T3-L1 adipocytes were more prone to palmitate-induced ER stress,indicating the involvement of Rab18 in alleviating lipid toxicity.Rab18 AKO mice provide a good animal model to investigate metabolic disorders such as impaired SAT.In conclusion,our studies reveal that Rab18 is a key and specific regulator that maintains the proper functions of SAT by alleviating lipid-induced ER stress.展开更多
Investigating correlations between radiomic and genomic profiling in breast cancer(BC)molecular subtypes is crucial for understanding disease mechanisms and providing personalized treatment.We present a well-designed ...Investigating correlations between radiomic and genomic profiling in breast cancer(BC)molecular subtypes is crucial for understanding disease mechanisms and providing personalized treatment.We present a well-designed radiogenomic framework image–gene–gene set(IMAGGS),which detects multi-way associations in BC subtypes by integrating radiomic and genomic features.Our dataset consists of 721 patients,each of whom has 12 ultrasound(US)images captured from different angles and gene mutation data.To better characterize tumor traits,12 multi-angle US images are fused using two distinct strategies.Then,we analyze complex many-to-many associations between phenotypic and genotypic features using a machine learning algorithm,deviating from the prevalent one-to-one relationship pattern observed in previous studies.Key radiomic and genomic features are screened using these associations.In addition,gene set enrichment analysis is performed to investigate the joint effects of gene sets and delve deeper into the biological functions of BC subtypes.We further validate the feasibility of IMAGGS in a glioblastoma multiforme dataset to demonstrate the scalability of IMAGGS across different modalities and diseases.Taken together,IMAGGS provides a comprehensive characterization for diseases by associating imaging,genes,and gene sets,paving the way for biological interpretation of radiomics and development of targeted therapy.展开更多
Glioblastoma multiforme(GBM)is the most malignant intracranial tumor in adults and its unique pathology leads to limited therapeutic benefits.1,2 Mitochondrial fusion and fission play an important role in carcinogenes...Glioblastoma multiforme(GBM)is the most malignant intracranial tumor in adults and its unique pathology leads to limited therapeutic benefits.1,2 Mitochondrial fusion and fission play an important role in carcinogenesis;fragmented mitochondria promote tumor cell proliferation and prolonged mitochondria lead to tumor cell apoptosis.3 Therefore,profiling the function and prognostic value of mitochondrial dynamics-related genes(MDRGs)is of great interest for GBM precision treatment.Here we focused on the expression,function,and genetic alterations of MDRGs and identified new DNA methylation sites being significantly associated with the survival of GBM patients using available data in public databases.展开更多
Tetralogy of Fallot(TOF)is the most common cyanotic congenital heart disease and the incidence of late cardiac death in long-term survivors continues to increase.1 So,there is an urgent need to explore the etiology an...Tetralogy of Fallot(TOF)is the most common cyanotic congenital heart disease and the incidence of late cardiac death in long-term survivors continues to increase.1 So,there is an urgent need to explore the etiology and pathogenesis of TOF.The precise cause of TOF is currently unclear,and exploration of the pathogenesis has focused increasingly in recent years on the roles of noncoding gene products,especially long noncoding RNAs(lncRNAs).展开更多
Metabolic homeostasis is regulated by a network of organs and tissues,primarily involving adipose tissue,muscle,liver,and the hypothalamus,which act as central metabolic regulators.Cellular dysregulation within these ...Metabolic homeostasis is regulated by a network of organs and tissues,primarily involving adipose tissue,muscle,liver,and the hypothalamus,which act as central metabolic regulators.Cellular dysregulation within these tissues substantially associates with metabolic disorders,including obesity,type 2 diabetes,and non-alcoholic fatty liver disease(NAFLD)[1].Understanding the molecular mechanisms governing metabolic control requires dedicated analysis of physiological and pathological cellular heterogeneity within these tissues.However,investigations at the single cell level to decipher the complexities of cellular mechanisms remain challenging due to the fragile nature of certain cell types and technical noise within these metabolically active tissues,resulting in limited studies compared to well-characterized atlases in immune cell populations[2].展开更多
Both microbiota and carcinogens play crucial roles in tumorigenesis,yet the effect of microbes on carcinogen biotransformation remains poorly understood.Roje et al.(2024)provide new insights into microbe-carcinogen in...Both microbiota and carcinogens play crucial roles in tumorigenesis,yet the effect of microbes on carcinogen biotransformation remains poorly understood.Roje et al.(2024)provide new insights into microbe-carcinogen interactions,demonstrating that intestinal microbiota drives bladder cancer development by modulating the metabolism of the carcinogen N-butyl-N-(4-hydroxybutyl)-nitrosamine(BBN).展开更多
The intestinal lymphatic system is essential for lipid absorption, yet its regulatory mechanisms remain poorly understood. Here, we identify DHHC5, an Asp-His-His-Cys (DHHC) motif-containing palmitoyl acyltransferase,...The intestinal lymphatic system is essential for lipid absorption, yet its regulatory mechanisms remain poorly understood. Here, we identify DHHC5, an Asp-His-His-Cys (DHHC) motif-containing palmitoyl acyltransferase, as a critical regulator of intestinal lymphatic integrity and lipid uptake. Whole-body inducible Dhhc5 knockout (Dhhc5-IKO) mice were resistant to diet-induced obesity and exhibited impaired intestinal lipid absorption due to lymphatic dysfunction. Similar defects were observed upon specific knockout of DHHC5 in lymphatic endothelial cells (LECs), underscoring its cell-autonomous role. Mechanistically, DHHC5 facilitates vascular endothelial growth factor receptor 2 (VEGFR2) signaling by promoting its lipid raft localization in LECs. We further identified CRYBG1, an actin-binding protein, as the substrate of DHHC5. CRYBG1 interacts with VEGFR2, and its palmitoylation is required for the lipid raft localization of VEGFR2. These findings reveal a DHHC5–CRYBG1–VEGFR2 axis that governs intestinal lymphatic function and lipid absorption, providing new insights into the regulation of dietary lipid metabolism.展开更多
Dear Editor,The T-box transcription factor T(TBXT,T,Brachyuary)gene has been identified as a tissue-specific transcriptional factor in vertebrates,regulating mesoderm formation and notochord differentiation during emb...Dear Editor,The T-box transcription factor T(TBXT,T,Brachyuary)gene has been identified as a tissue-specific transcriptional factor in vertebrates,regulating mesoderm formation and notochord differentiation during embryonic development(Kispert et al.,1995).Tbxt knockout heterozygous mice exhibited varying degrees of tail shortening,while homozygous mice died at approximately E10.0,and displayed severe defects in somite formation and neural tube development(Abe et al.,2000).展开更多
Obesity is a major pathological factor that induces insulin resistance and consequent type 2 diabetes through multiple mechanisms.Inactivation of the insulin receptor(INSR)contributes to the development of insulin res...Obesity is a major pathological factor that induces insulin resistance and consequent type 2 diabetes through multiple mechanisms.Inactivation of the insulin receptor(INSR)contributes to the development of insulin resistance,whose protein level is down-regulated in obesity through as yet-undefined mechanisms.Here we show that the E3-ligase TRAF6 is a critical regulator of INSR maturation,whose inactivation prevents palmitic acid-or high-fat diet-induced diminution of the INSR.Consequently,genetic inactivation of TRAF6 enhances insulin signaling that further increases muscle glucose uptake and inhibits hepatic gluconeogenesis.TRAF6 inactivation increases the proprotein convertase FURIN that controls the processing of pro-INSR to mature INSR.Mechanistically,TRAF6 associates with the Golgi apparatus,where it ubiquitinates the cytosolic tail of FURIN,leading to its lysosomal degradation.This TRAF6-FURIN axis also regulates cholesterol metabolism via PCSK9 processing in the circulation.Collectively,our results reveal a critical role of TRAF6 in regulating proprotein processing and have therapeutic implications for metabolic control.展开更多
Brown adipose tissue (BAT) plays an essential role in non-shivering thermogenesis. The phosphatidylinositol transfer protein, cytoplasmic 1 (PITPNC1) is identified as a lipid transporter that reciprocally transfers ph...Brown adipose tissue (BAT) plays an essential role in non-shivering thermogenesis. The phosphatidylinositol transfer protein, cytoplasmic 1 (PITPNC1) is identified as a lipid transporter that reciprocally transfers phospholipids between intracellular membrane structures. However, the physiological significance of PITPNC1 and its regulatory mechanism remain unclear. Here, we demonstrate that PITPNC1 is a key player in thermogenesis of BAT. While Pitpnc1^(−/−) mice do not differ with wildtype mice in body weight and insulin sensitivity on either chow or high-fat diet, they develop hypothermia when subjected to acute cold exposure at 4℃. The Pitpnc1^(−/−) brown adipocytes exhibit defective β-oxidation and abnormal thermogenesis-related metabolism pathways in mitochondria. The deficiency of lipid mobilization in Pitpnc1^(−/−) brown adipocytes might be the result of excessive accumulation of phosphatidylcholine and a reduction of phosphatidic acid. Our findings have uncovered significant roles of PITPNC1 in mitochondrial phospholipid homeostasis and BAT thermogenesis.展开更多
Brown adipocyte maturation during postnatal development is essential for brown adipose tissue(BAT)to protect animals against cold.Impaired maturation of brown adipocytes leads to cold intolerance.However,the molecular...Brown adipocyte maturation during postnatal development is essential for brown adipose tissue(BAT)to protect animals against cold.Impaired maturation of brown adipocytes leads to cold intolerance.However,the molecular mechanisms that determine the maturation of brown adipocytes during postnatal development are not fully understood.Here,we identify Wilms’tumor 1-associating protein(WTAP)as an essential regulator in the postnatal development and maturation of BAT.BAT-specific knockout of Wtap(Wtap-BKO)severely impairs maturation of BAT in vivo by decreasing the expression of BAT-selective genes,leading to the whitening of interscapular BAT(iBAT).Single nucleus RNA-sequencing analysis shows the dynamic changes of cell heterogeneity in iBAT of Wtap-BKO mice.Adult mice with WTAP deficiency in BAT display hypothermic and succumb to acute cold challenge.Mechanistically,WTAP deficiency decreases m6A mRNA modification by reducing the protein stability of METTL3.BAT-specific overexpression of Mettl3 partially rescues the phenotypes observed in Wtap-BKO mice.These data demonstrate that WTAP/METTL3 plays an essential role in iBAT postnatal development and thermogenesis.展开更多
It is widely recognized that tumor immune microenvironment(TIME)plays a crucial role in tumor progression,metastasis,and therapeutic response.Despite several noninvasive strategies have emerged for cancer diagnosis an...It is widely recognized that tumor immune microenvironment(TIME)plays a crucial role in tumor progression,metastasis,and therapeutic response.Despite several noninvasive strategies have emerged for cancer diagnosis and prognosis,there are still lack of efective radiomic-based model to evaluate TIME status,let alone predict clinical outcome and immune checkpoint inhibitor(ICIs)response for hepatocellular carcinoma(HCC).In this study,we developed a radiomic model to evaluate TIME status within the tumor and predict prognosis and immunotherapy response.A total of 301 patients who underwent magnetic resonance imaging(MRI)examinations were enrolled in our study.The intra-tumoral expression of 17 immune-related molecules were evaluated using co-detection by indexing(CODEX)technology,and we construct Immunoscore(IS)with the least absolute shrinkage and selection operator(LASSO)algorithm and Cox regression method to evaluate TIME.Of 6115 features extracted from MRI,fve core features were fltered out,and the Radiomic Immunoscore(RIS)showed high accuracy in predicting TIME status in testing cohort(area under the curve=0.753).More importantly,RIS model showed the capability of predicting therapeutic response to anti-programmed cell death 1(PD-1)immunotherapy in an independent cohort with advanced HCC patients(area under the curve=0.731).In comparison with previously radiomicbased models,our integrated RIS model exhibits not only higher accuracy in predicting prognosis but also the potential guiding signifcance to HCC immunotherapy.展开更多
基金This study was supported by the National Key Research and Development Program of China(2021YFC2701400 and 2023YFC2705600)the National Natural Science Foundation of China(32288101,32100480,32370654,82271639,32322017,and32200485).
文摘Oligoasthenoteratozoospermia is an important factor affecting male fertility and has been found to be associated with genetic factors.However,there are stll a proportion of oligoasthenoteratozoospermia cases that cannot be explained by known pathogenic genetic variants.Here,we perform genetic analyses and identify bi-allelic loss-of-function variants of MFSD6L from an oligoasthenoteratozoospermia-affected family.Mfsd6l knock-out male mice also present male subfertility with reduced sperm concentration,motility,and deformed acrosomes.Further mechanistic analyses reveal that MFsD6L,as an acrosome membrane protein,plays an important role in the formation of acrosome by interacting with the inner acrosomal membrane protein SPACA1.Moreover,poor embryonic development is consistently observed after intracytoplasmic sperm injection treatment using spermatozoa from the MFSD6L-deficient man and male mice.Collectively,our findings reveal that MFSD6L is required for the anchoring of sperm acrosome and head shaping.The deficiency of MFsD6L affects male fertility and causes oligoasthenoter-atozoospermia in humans and mice.
基金supported by the National Key Research and Development Program of China(2021YFC2701400)in part by the National Natural Science Foundation of China(32000393 and 32288101).
文摘During spermiogenesis,haploid spermatids undergo dramatic morphological changes to form slender sperm flagella and cap-like acrosomes,which are required for successful fertilization.Severe deformities in flagella cause a male infertility syndrome,multiple morphological abnormalities of the flagella(MMAF),while acrosomal hypoplasia in some cases leads to sub-optimal embryonic developmental potential.However,evidence regarding the occurrence of acrosomal hypoplasia in MMAF is limited.Here,we report the generation of base-edited mice knocked out for coiled-coil domain-containing 38(Ccdc38)via inducing a nonsense mutation and find that the males are infertile.The Ccdc38-KO sperm display acrosomal hypoplasia and typical MMAF phenotypes.We find that the acrosomal membrane is loosely anchored to the nucleus and fibrous sheaths are disorganized in Ccdc38-KO sperm.Further analyses reveal that Ccdc38 knockout causes a decreased level of TEKT3,a protein associated with acrosome biogenesis,in testes and an aberrant distribution of TEKT3 in sperm.We finally show that intracytoplasmic sperm injection overcomes Ccdc38-related infertility.Our study thus reveals a previously unknown role for CCDC38 in acrosome biogenesis and provides additional evidence for the occurrence of acrosomal hypoplasia in MMAF.
基金supported by the National Key R&D Program of China(2021YFC2701101 to H.W.and X.Y.)the National Natural Science Foundation of China(81930036 and 82150008 to H.W.,and 31000542 to X.Y.)the Commission of Science and Technology of Shanghai Municipality(20JC1418500 to H.W.).
文摘T-box transcription factor T(TBXT;T)is required for mesodermal formation and axial skeletal development.Although it has been extensively studied in various model organisms,human congenital vertebral malformations(CVMs)involving T are not well established.Here,we report a family with 15 CVM patients distributed across 4 generations.All affected individuals carry a heterozygous mutation,T c.596A>G(p.Q199R),which is not found in unaffected family members,indicating co-segregation of the genotype and phenotype.In vitro assays show that T p.Q199R increases the nucleocytoplasmic ratio and enhances its DNA-binding affinity,but reduces its transcriptional activity compared to the wild-type.To determine the pathogenicity of this mutation in vivo,we generated a Q199R knock-in mouse model that recapitulates the human CVM phenotype.Most heterozygous Q199R mice show subtle kinked or shortened tails,while homozygous mice exhibit tail filaments and severe vertebral deformities.Overall,we show that the Q199R mutation in T causes CVM in humans and mice,providing previously unreported evidence supporting the function of T in the genetic etiology of human CVM.
基金supported by grants from the National Key R&D Program of China(2020YFA0803800(Ru W.),2019YFA0801900,2018YFA0800300(T.L.)the National Natural Science Foundation of China(31971097,32271226(Ru W.),92357304,92249302,32150610475,31971074(T.L.),82200952(Q.Z.),32271191(Y.T.),81800751(Q.S.),82100584(Y.S.),82000790(W.W.),82200949(L.F.),81970458(Q.Y.))+3 种基金Shanghai“Science and Technology Innovation Action Plan”Social Development Science and Technology Reach Project(22dz1204600)(Ru W.)Shanghai Municipal Science and Technology Committee of Shanghai outstanding academic leaders plan(21XD1403200)(Ru W.)the Construction Project of High-Level Local Universities in Shanghai,China,Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(ZYYCXTD-D-202001)(T.L.)the Yangfan Project of Shanghai Science and Technology Commission(21YF1405000)(Y.F.)。
文摘Sedentary behavior for two years during the coronavirus disease 2019(COVID-19)pandemic contributes to weight gain.Gut microbiota and blood metabolome are related to body mass index(BMI)and indicate individual metabolic changes.Surgery and exercise are effective weight-loss methods.The precise plasma metabolites and gut microbiota biomarkers involved and the underlying mechanisms are still largely unclear.To address this issue,we analyzed weight gain and weight loss cohorts to identify biomarkers associated with obesity.In the sedentary cohort,49 subjects were recruited in year 2019.After two years of sedentary behavior during the COVID-19 pandemic,the BMI of 24 subjects significantly increased(Weight gain group),while that of the remnant 25 subjects remained constant(Maintaining weight group).At baseline and two years post baseline,the gut microbiota and blood metabolome,as well as body composition and clinical indicators,were all collected.In weight loss studies,we analyze the plasma metabolome of the two cohorts,including individuals who underwent laparoscopic sleeve gastrectomy(LSG)surgery and exercise intervention.Weight gain through sedentary behavior contributed to the variation of the gut microbiota and plasma metabolites composition.Creatine,phenylalanine and tyrosine exhibited significant positive associations with BMI and fat mass.We further confirmed the association between BMI and plasma metabolites in two weight loss cohorts.By utilizing a linear regression model,we found that 10 metabolites including creatine were correlated with BMI in weight loss individuals.Based on receiver operating characteristic(ROC)curves,creatine exhibited a satisfactory classification performance in regard to predicting weight reduction(AUC_(LSG)=0.890,AUC_(Sports)=0.840).Moreover,some gut microbiota,including Bifidobacterium angulatum DSM 20098=JCM 7096 and Rothia dentocariosa M567I could affect BMI through the mediating factor of creatine.
基金supported by the National Natural Science Foundation of China(82273223 to F.L.,32270798 to P.L.)the National Key Research and Development Program of China(2022YFA1103900 to F.L.).
文摘Bromodomain and extra-terminal domain(BET)proteins,which function partly through MYC proto-oncogene(MYC),are critical epigenetic readers and emerging therapeutic targets in cancer.Whether and how BET inhibition simultaneously induces metabolic remodeling in cancer cells remains unclear.Here we find that even transient BET inhibition by JQ-1 and other pan-BET inhibitors(pan-BETis)blunts liver cancer cell proliferation and tumor growth.BET inhibition decreases glycolytic gene expression but enhances mitochondrial glucose and glutamine oxidative metabolism revealed by metabolomics and isotope labeling analysis.Specifically,BET inhibition downregulates miR-30a to upregulate glutamate dehydrogenase 1(GDH1)independent of MYC,which producesα-ketoglutarate for mitochondrial oxidative phosphorylation(OXPHOS).Targeting GDH1 or OXPHOS is synthetic lethal to BET inhibi-tion,and combined BET and OXPHOS inhibition therapeutically prevents liver tumor growth in vitro and in vivo.Together,we uncover an important epigenetic-metabolic crosstalk whereby BET inhibition induces MYC-independent and GDH1-dependent glutamine metabolic remodeling that can be exploited for innovative combination therapy of liver cancer.
基金supported by the National Key R&D Program of China(2019YFA0801900,2018YFA0800300)the National Natural Science Foundation of China(31971074)+3 种基金the Science and Technology Innovation Action Plan of Shanghai Science and Technology Committee(18140901300)the Open Research Fund of the National key laboratory of genetic engineering(SKLGE1803)the Shanghai Municipal Science and Technology Major Project(2017SHZDZX01)Shanghai Frontiers Science Research Base of Exercise and Metabolic Health.
文摘βcells are defined by the ability to produce and secret insulin.Recent studies have evaluated that human pancreaticβcells are heterogeneous and demonstrated the transcript alterations ofβcell subpopulation in diabetes.Single-cell RNA sequence(scRNA-seq)analysis helps us to refine the cell types signatures and understand the role of theβcells during metabolic challenges and diseases.Here,we construct the pseudotime trajectory ofβcells from publicly available scRNA-seq data in health and type 2 diabetes(T2D)based on highly dispersed and highly expressed genes using Monocle2.We identified three major states including 1)Normal branch,2)Obesity-like branch and 3)T2D-like branch based on biomarker genes and genes that give rise to bifurcation in the trajectory.βcell function-maintain-related genes,insulin expression-related genes,and T2D-related genes enriched in three branches,respectively.Continuous pseudotime spectrum might suggest thatβcells transition among different states.The application of pseudotime analysis is conducted to clarify the different cell states,providing novel insights into the pathology ofβcells in T2D.
基金supported by the National Key Research and Development Program of China(2021YFC2701400)the National Natural Science Foundation of China(32000393,32322017,32288101)。
文摘Cytosine base editing achieves C·G-to-T·A substitutions and can convert four codons(CAA/CAG/CGA/TGG)into STOP-codons(induction of STOP-codons,iSTOP)to knock out genes with reduced mosaicism.iSTOP enables direct phenotyping in founders’somatic cells,but it remains unknown whether this works in founders’germ cells so as to rapidly reveal novel genes for fertility.Here,we initially establish that iSTOP in mouse zygotes enables functional characterization of known genes in founders’germ cells:Cfap43-iSTOP male founders manifest expected sperm features resembling human“multiple morphological abnormalities of the flagella”syndrome(i.e.,MMAF-like features),while oocytes of Zp3-iSTOP female founders have no zona pellucida.We further illustrate iSTOP’s utility for dissecting the functions of unknown genes with Ccdc183,observing MMAF-like features and male infertility in Ccdc183-iSTOP founders,phenotypes concordant with those of Ccdc183-KO offspring.We ultimately establish that CCDC183 is essential for sperm morphogenesis through regulating the assembly of outer dynein arms and participating in the intra-flagellar transport.Our study demonstrates iSTOP as an efficient tool for direct reproductive disease modeling and phenotyping in germ cells of the founder generation,and rapidly reveals the essentiality of Ccdc183 in fertility,thus providing a time-saving approach for validating genetic defects(like nonsense mutations)for human infertility.
基金supported by grants from the National Key R&D Program of China(2021YFC2701101,H.W.)the National Natural Science Foundation of China(81930036 and 82150008,H.W.)the Commission for Science and Technology of Shanghai Municipality(20JC1418500,H.W.).
文摘Dear Editor,Tetralogy of Fallot(TOF)is the most common complex congenital heart disease.Besides gene mutations and copy number variants,altered protein function induced by posttranscriptional or translational regulation also contributes to the onset of TOF.1 MiRNAs are short noncoding RNAs that bind to the 3’-UTR of target mRNAs to repress protein production.However,the causal link between miRNAs and TOF and the underlying mechanism has not been established.
基金supported by the National Key Research and Development Program of China(2018YFA0506901,2019YFA0801701,2022YFA0806502)the National Natural Science Foundation of China(92254308,92157107)the Lingang Laboratory(LG-QS-202204-06)。
文摘Metabolically healthy obesity refers to obese individuals who do not develop metabolic disorders.These people store fat in subcutaneous adipose tissue(SAT)rather than in visceral adipose tissue(VAT).However,the molecules participating in this specific scenario remain elusive.Rab18,a lipid droplet(LD)-associated protein,mediates the contact between the endoplasmic reticulum(ER)and LDs to facilitate LD growth and maturation.In the present study,we show that the protein level of Rab18 is specifically upregulated in the SAT of obese people and mice.Rab18 adipocyte-specific knockout(Rab18 AKO)mice had a decreased volume ratio of SAT to VAT compared with wildtype mice.When subjected to high-fat diet(HFD),Rab18 AKO mice had increased ER stress and inflammation,reduced adiponectin,and decreased triacylglycerol(TAG)accumulation in SAT.In contrast,TAG accumulation in VAT,brown adipose tissue(BAT)or liver of Rab18AKO mice had a moderate increase without ER stress stimulation.Rab18 AKO mice developed insulin resistance and systematic inflammation.Rab18 AKO mice maintained body temperature in response to acute and chronic cold induction with a thermogenic SAT,similar to the counterpart mice.Furthermore,Rab18-deficient 3T3-L1 adipocytes were more prone to palmitate-induced ER stress,indicating the involvement of Rab18 in alleviating lipid toxicity.Rab18 AKO mice provide a good animal model to investigate metabolic disorders such as impaired SAT.In conclusion,our studies reveal that Rab18 is a key and specific regulator that maintains the proper functions of SAT by alleviating lipid-induced ER stress.
基金supported by the National Natural Science Foundation of China(81830058,82071945,91959207,92159301,and 82302212)the Science and Technology Commission of Shanghai Municipality(22ZR1404800)。
文摘Investigating correlations between radiomic and genomic profiling in breast cancer(BC)molecular subtypes is crucial for understanding disease mechanisms and providing personalized treatment.We present a well-designed radiogenomic framework image–gene–gene set(IMAGGS),which detects multi-way associations in BC subtypes by integrating radiomic and genomic features.Our dataset consists of 721 patients,each of whom has 12 ultrasound(US)images captured from different angles and gene mutation data.To better characterize tumor traits,12 multi-angle US images are fused using two distinct strategies.Then,we analyze complex many-to-many associations between phenotypic and genotypic features using a machine learning algorithm,deviating from the prevalent one-to-one relationship pattern observed in previous studies.Key radiomic and genomic features are screened using these associations.In addition,gene set enrichment analysis is performed to investigate the joint effects of gene sets and delve deeper into the biological functions of BC subtypes.We further validate the feasibility of IMAGGS in a glioblastoma multiforme dataset to demonstrate the scalability of IMAGGS across different modalities and diseases.Taken together,IMAGGS provides a comprehensive characterization for diseases by associating imaging,genes,and gene sets,paving the way for biological interpretation of radiomics and development of targeted therapy.
基金the Key R&D Program of the Science and Technology Ministry of China(No.2021YFC2701100)the National Natural Science Foundation of China(No.82150008,81930036)the Commission of Science and Technology of Shanghai Municipality,China(No.20JC1418500).
文摘Glioblastoma multiforme(GBM)is the most malignant intracranial tumor in adults and its unique pathology leads to limited therapeutic benefits.1,2 Mitochondrial fusion and fission play an important role in carcinogenesis;fragmented mitochondria promote tumor cell proliferation and prolonged mitochondria lead to tumor cell apoptosis.3 Therefore,profiling the function and prognostic value of mitochondrial dynamics-related genes(MDRGs)is of great interest for GBM precision treatment.Here we focused on the expression,function,and genetic alterations of MDRGs and identified new DNA methylation sites being significantly associated with the survival of GBM patients using available data in public databases.
基金supported by grants from the National Key R&D Program of China(No.2021YFC2701101)the National Natural Science Foundation of China(No.81930036 and 82150008)the Commission for Science and Technology of Shanghai Municipality(China)(No.20ZR1404800).
文摘Tetralogy of Fallot(TOF)is the most common cyanotic congenital heart disease and the incidence of late cardiac death in long-term survivors continues to increase.1 So,there is an urgent need to explore the etiology and pathogenesis of TOF.The precise cause of TOF is currently unclear,and exploration of the pathogenesis has focused increasingly in recent years on the roles of noncoding gene products,especially long noncoding RNAs(lncRNAs).
基金funded by the National Key Research and Development Program of China (2024YFA1802800 and 2024YFA1802803)the National Natural Science Foundation of China (32371195)a new PI Start up grant of Fudan University (JIH2303132Y) to G.W
文摘Metabolic homeostasis is regulated by a network of organs and tissues,primarily involving adipose tissue,muscle,liver,and the hypothalamus,which act as central metabolic regulators.Cellular dysregulation within these tissues substantially associates with metabolic disorders,including obesity,type 2 diabetes,and non-alcoholic fatty liver disease(NAFLD)[1].Understanding the molecular mechanisms governing metabolic control requires dedicated analysis of physiological and pathological cellular heterogeneity within these tissues.However,investigations at the single cell level to decipher the complexities of cellular mechanisms remain challenging due to the fragile nature of certain cell types and technical noise within these metabolically active tissues,resulting in limited studies compared to well-characterized atlases in immune cell populations[2].
基金supported by the National Key R&D Program of China(2022YFA0806400,2021YFC2701100)National Natural Science Foundation of China(32271215)。
文摘Both microbiota and carcinogens play crucial roles in tumorigenesis,yet the effect of microbes on carcinogen biotransformation remains poorly understood.Roje et al.(2024)provide new insights into microbe-carcinogen interactions,demonstrating that intestinal microbiota drives bladder cancer development by modulating the metabolism of the carcinogen N-butyl-N-(4-hydroxybutyl)-nitrosamine(BBN).
基金supported by the National Natural Science Foundation of China(32125022,32230053,92157301,and 32401059)the National Key R&D Program of China(2024YFA1306101 and 2020YFA0803601)the Shanghai Basic Research Field Project“Science and Technology Innovation Action Plan”(21JC1400400).
文摘The intestinal lymphatic system is essential for lipid absorption, yet its regulatory mechanisms remain poorly understood. Here, we identify DHHC5, an Asp-His-His-Cys (DHHC) motif-containing palmitoyl acyltransferase, as a critical regulator of intestinal lymphatic integrity and lipid uptake. Whole-body inducible Dhhc5 knockout (Dhhc5-IKO) mice were resistant to diet-induced obesity and exhibited impaired intestinal lipid absorption due to lymphatic dysfunction. Similar defects were observed upon specific knockout of DHHC5 in lymphatic endothelial cells (LECs), underscoring its cell-autonomous role. Mechanistically, DHHC5 facilitates vascular endothelial growth factor receptor 2 (VEGFR2) signaling by promoting its lipid raft localization in LECs. We further identified CRYBG1, an actin-binding protein, as the substrate of DHHC5. CRYBG1 interacts with VEGFR2, and its palmitoylation is required for the lipid raft localization of VEGFR2. These findings reveal a DHHC5–CRYBG1–VEGFR2 axis that governs intestinal lymphatic function and lipid absorption, providing new insights into the regulation of dietary lipid metabolism.
基金supported by the Basic Science Center Program(32488101)Shenzhen Medical Research Fund(B2402004)+1 种基金Sanming Project of Medicine in Shenzhen(SZSM202311005)Shanghai Municipal Science and Technology Major Project(FCK-92724413-2024-025)。
文摘Dear Editor,The T-box transcription factor T(TBXT,T,Brachyuary)gene has been identified as a tissue-specific transcriptional factor in vertebrates,regulating mesoderm formation and notochord differentiation during embryonic development(Kispert et al.,1995).Tbxt knockout heterozygous mice exhibited varying degrees of tail shortening,while homozygous mice died at approximately E10.0,and displayed severe defects in somite formation and neural tube development(Abe et al.,2000).
基金supported by the National Natural Science Foundation of China(32025019)the Ministry of Science and Technology of China(2021YFF0702100,2018YFA0801100)+2 种基金the Science and Technology Foundation of Jiangsu Province of China(BK20243041)the Fundamental Research Funds for the Central Universities(021414380533)Wellcome Trust Investigator Award(209380/Z/17/Z).
文摘Obesity is a major pathological factor that induces insulin resistance and consequent type 2 diabetes through multiple mechanisms.Inactivation of the insulin receptor(INSR)contributes to the development of insulin resistance,whose protein level is down-regulated in obesity through as yet-undefined mechanisms.Here we show that the E3-ligase TRAF6 is a critical regulator of INSR maturation,whose inactivation prevents palmitic acid-or high-fat diet-induced diminution of the INSR.Consequently,genetic inactivation of TRAF6 enhances insulin signaling that further increases muscle glucose uptake and inhibits hepatic gluconeogenesis.TRAF6 inactivation increases the proprotein convertase FURIN that controls the processing of pro-INSR to mature INSR.Mechanistically,TRAF6 associates with the Golgi apparatus,where it ubiquitinates the cytosolic tail of FURIN,leading to its lysosomal degradation.This TRAF6-FURIN axis also regulates cholesterol metabolism via PCSK9 processing in the circulation.Collectively,our results reveal a critical role of TRAF6 in regulating proprotein processing and have therapeutic implications for metabolic control.
基金the National Key R&D Program of China(2018YFA0506900)the National Key R&D Program of China(2018YFA0800301)+3 种基金the National Natural Science Foundation of China(91857103)Shanghai Basic Research Field Project“Science and Technology Innovation Action Plan”(21JC1400400)the Lingang Laboratory(LG-QS-202204-06)Shanghai Municipal Science and Technology Major Project(2017SHZDZX01)。
文摘Brown adipose tissue (BAT) plays an essential role in non-shivering thermogenesis. The phosphatidylinositol transfer protein, cytoplasmic 1 (PITPNC1) is identified as a lipid transporter that reciprocally transfers phospholipids between intracellular membrane structures. However, the physiological significance of PITPNC1 and its regulatory mechanism remain unclear. Here, we demonstrate that PITPNC1 is a key player in thermogenesis of BAT. While Pitpnc1^(−/−) mice do not differ with wildtype mice in body weight and insulin sensitivity on either chow or high-fat diet, they develop hypothermia when subjected to acute cold exposure at 4℃. The Pitpnc1^(−/−) brown adipocytes exhibit defective β-oxidation and abnormal thermogenesis-related metabolism pathways in mitochondria. The deficiency of lipid mobilization in Pitpnc1^(−/−) brown adipocytes might be the result of excessive accumulation of phosphatidylcholine and a reduction of phosphatidic acid. Our findings have uncovered significant roles of PITPNC1 in mitochondrial phospholipid homeostasis and BAT thermogenesis.
基金This study was supported by the National Natural Science Foundation of China grant(92057110,31971083,92057115,and 32071138)National Key R&D Program of China(2020YFA0803800,2019YFA0801900,2020YFA0803601,and 2018YFA0801300).
文摘Brown adipocyte maturation during postnatal development is essential for brown adipose tissue(BAT)to protect animals against cold.Impaired maturation of brown adipocytes leads to cold intolerance.However,the molecular mechanisms that determine the maturation of brown adipocytes during postnatal development are not fully understood.Here,we identify Wilms’tumor 1-associating protein(WTAP)as an essential regulator in the postnatal development and maturation of BAT.BAT-specific knockout of Wtap(Wtap-BKO)severely impairs maturation of BAT in vivo by decreasing the expression of BAT-selective genes,leading to the whitening of interscapular BAT(iBAT).Single nucleus RNA-sequencing analysis shows the dynamic changes of cell heterogeneity in iBAT of Wtap-BKO mice.Adult mice with WTAP deficiency in BAT display hypothermic and succumb to acute cold challenge.Mechanistically,WTAP deficiency decreases m6A mRNA modification by reducing the protein stability of METTL3.BAT-specific overexpression of Mettl3 partially rescues the phenotypes observed in Wtap-BKO mice.These data demonstrate that WTAP/METTL3 plays an essential role in iBAT postnatal development and thermogenesis.
基金National Natural Science Foundation of China(U21A20376,81988101,81790633 and 81830054)National Science Foundation of Shanghai(21XD1404600,17ZR143800,21JC1406600 and 22140901000)Project of Shanghai Municipal Commission of Health(2022LJ024).
文摘It is widely recognized that tumor immune microenvironment(TIME)plays a crucial role in tumor progression,metastasis,and therapeutic response.Despite several noninvasive strategies have emerged for cancer diagnosis and prognosis,there are still lack of efective radiomic-based model to evaluate TIME status,let alone predict clinical outcome and immune checkpoint inhibitor(ICIs)response for hepatocellular carcinoma(HCC).In this study,we developed a radiomic model to evaluate TIME status within the tumor and predict prognosis and immunotherapy response.A total of 301 patients who underwent magnetic resonance imaging(MRI)examinations were enrolled in our study.The intra-tumoral expression of 17 immune-related molecules were evaluated using co-detection by indexing(CODEX)technology,and we construct Immunoscore(IS)with the least absolute shrinkage and selection operator(LASSO)algorithm and Cox regression method to evaluate TIME.Of 6115 features extracted from MRI,fve core features were fltered out,and the Radiomic Immunoscore(RIS)showed high accuracy in predicting TIME status in testing cohort(area under the curve=0.753).More importantly,RIS model showed the capability of predicting therapeutic response to anti-programmed cell death 1(PD-1)immunotherapy in an independent cohort with advanced HCC patients(area under the curve=0.731).In comparison with previously radiomicbased models,our integrated RIS model exhibits not only higher accuracy in predicting prognosis but also the potential guiding signifcance to HCC immunotherapy.
