生长因子受体结合蛋白10(Growth factor receptor-bound protein 10,Grb10)是一个存在于小鼠11号染色体和人7号染色体的母本表达的印记基因。文章利用原位杂交技术和定量RT-PCR方法对不同发育阶段的小鼠胚胎Grb10基因进行时空表达谱的分...生长因子受体结合蛋白10(Growth factor receptor-bound protein 10,Grb10)是一个存在于小鼠11号染色体和人7号染色体的母本表达的印记基因。文章利用原位杂交技术和定量RT-PCR方法对不同发育阶段的小鼠胚胎Grb10基因进行时空表达谱的分析,以确定该基因在胚胎发育中其表达与组织发育的关系。定量RT-PCR数据结果表明,Grb10在E8.5-E13.5(Embryonic days8.5-13.5),表达水平逐渐增高,在E13.5达到高峰,后期表达量回落。在胚胎发育的中后期脑、心、肺组织的表达水平呈递减趋势。肝脏组织中Grb10表达水平较为恒定,在E18.5出现高峰。原位杂交数据验证了定量RT-PCR的结果,并且说明了Grb10在其他如骨、肾和肌肉等组织器官的高表达水平。研究结果表明Grb10基因是小鼠胚期发育的一个重要的基因。展开更多
Decreased functional β-cell mass is the hallmark of diabetes, but the cause of this metabolic defect remains elusive. Here, we show that the levels of the growth factor receptor-bound protein 10(GRB10), a negative re...Decreased functional β-cell mass is the hallmark of diabetes, but the cause of this metabolic defect remains elusive. Here, we show that the levels of the growth factor receptor-bound protein 10(GRB10), a negative regulator of insulin and m TORC1 signaling, are markedly induced in islets of diabetic mice and high glucose-treated insulinoma cell line INS-1 cells. β-cell-specific knockout of Grb10 in mice increased β-cell mass and improved β-cell function. Grb10-deficient β-cells exhibit enhanced m TORC1 signaling and reduced β-cell dedifferentiation, which could be blocked by rapamycin. On the contrary, Grb10 overexpression induced β-cell dedifferentiation in MIN6 cells. Our study identifies GRB10 as a critical regulator of β-cell dedifferentiation and β-cell mass, which exerts its effect by inhibiting m TORC1 signaling.展开更多
Domestic pigs are shaped by artificial and natural selection into obese and lean types that are closely related to muscle tissue.However,the key genes and regulatory mechanisms behind these developments remain largely...Domestic pigs are shaped by artificial and natural selection into obese and lean types that are closely related to muscle tissue.However,the key genes and regulatory mechanisms behind these developments remain largely unknown.Here,we pinpoint GRB10 specificity in muscle tissue and cells between obese and lean pigs by combining genomics,transcriptomics,epigenomics,and single-cell transcriptomics.GRB10 shows notable differences in divergent selection on haplotype blocks and expression levels between obese and lean pig breeds,with its expression profiles varying significantly by tissue and development stage.Notably,we identify a muscle-specific promoter of GRB10 and its transcription factor KLF15.This TF-promoter binding is verified by dual luciferase and chromatin immunoprecipitation(Ch IP)assays,and is suggested to be conserved in humans.Single-nucleus RNA sequencing further highlights differential expression patterns of GRB10 between obese and lean pig breeds across various cell types.Type IIa myonuclei and TTN+FAPs,which are more predominant in lean pigs,play a crucial role in myofibril assembly and muscle tissue development.These findings offer insights into the regulatory mechanisms controlling muscle growth.They highlight the tissue-and cell type-specific effects of GRB10 on muscle heterogeneity,which has potential applications in livestock breeding and human obesity research.展开更多
The development and homeostasis of intestinal epithelium are mediated by actively proliferating Lgr5^(+)stem cells,which pos-sess a remarkable self-renewal and differentiation capacity.Recently,our study demonstrated ...The development and homeostasis of intestinal epithelium are mediated by actively proliferating Lgr5^(+)stem cells,which pos-sess a remarkable self-renewal and differentiation capacity.Recently,our study demonstrated that N^(6)-methyladenosine(m^(6)A)methylation was essential for the survival of colonic stem cells.Here,we show that methyltransferase-like 3(METTL3)expression is downregulated in the colon mucosa in ulcerative colitis(UC)patients and strongly associated with the differentiation and maturation of goblet cells during inflammation.In mice,depletion of Mettl3 significantly inhibits the self-renewal and differentiation of Lgr5^(+)stem cells,especially the differentiation and maturation of goblet cells,resulting in intestinal dysplasia and spontaneous inflammation.Mechanistically,Mettl3 deletion-mediated m^(6)A loss facilitates the expression levels of growth factor receptor binding protein 10(Grb10)and interferon-related developmental regulator 1(Ifrd1)via increasing their messenger RNA stability.We further demonstrate that the levels of GRB10 and IFRD1 are negatively correlated with METTL3 level in UC samples.Collectively,our data indicate that METTL3 enhances the self-renewal and differentiation of Lgr5^(+)stem cells during intestinal development and inflammation,and thus it may be a potential therapeutic target for UC treatment.展开更多
基金supported by grants from the National Natural Science Foundation of China (91749118, 82070807, 81770775, 81730022)Natural Science Foundation of Hunan Province, China (2021JJ30976)National Key Research and Development Program (2019YFA0801903, 2018YFC2000100)。
文摘Decreased functional β-cell mass is the hallmark of diabetes, but the cause of this metabolic defect remains elusive. Here, we show that the levels of the growth factor receptor-bound protein 10(GRB10), a negative regulator of insulin and m TORC1 signaling, are markedly induced in islets of diabetic mice and high glucose-treated insulinoma cell line INS-1 cells. β-cell-specific knockout of Grb10 in mice increased β-cell mass and improved β-cell function. Grb10-deficient β-cells exhibit enhanced m TORC1 signaling and reduced β-cell dedifferentiation, which could be blocked by rapamycin. On the contrary, Grb10 overexpression induced β-cell dedifferentiation in MIN6 cells. Our study identifies GRB10 as a critical regulator of β-cell dedifferentiation and β-cell mass, which exerts its effect by inhibiting m TORC1 signaling.
基金supported by the National Natural Science Foundation of China(32172765)。
文摘Domestic pigs are shaped by artificial and natural selection into obese and lean types that are closely related to muscle tissue.However,the key genes and regulatory mechanisms behind these developments remain largely unknown.Here,we pinpoint GRB10 specificity in muscle tissue and cells between obese and lean pigs by combining genomics,transcriptomics,epigenomics,and single-cell transcriptomics.GRB10 shows notable differences in divergent selection on haplotype blocks and expression levels between obese and lean pig breeds,with its expression profiles varying significantly by tissue and development stage.Notably,we identify a muscle-specific promoter of GRB10 and its transcription factor KLF15.This TF-promoter binding is verified by dual luciferase and chromatin immunoprecipitation(Ch IP)assays,and is suggested to be conserved in humans.Single-nucleus RNA sequencing further highlights differential expression patterns of GRB10 between obese and lean pig breeds across various cell types.Type IIa myonuclei and TTN+FAPs,which are more predominant in lean pigs,play a crucial role in myofibril assembly and muscle tissue development.These findings offer insights into the regulatory mechanisms controlling muscle growth.They highlight the tissue-and cell type-specific effects of GRB10 on muscle heterogeneity,which has potential applications in livestock breeding and human obesity research.
基金supported by grants from the National Natural Science Foundation of China(82202017,32470951,82273214,and 82200119)the Shanghai Science and Technology Commission(22ZR1439600 and 20JC1410100)+2 种基金the Medical Talent Plan of High-level Local Universities in Shanghai(KJ3-0221-22-6338)Chongqing Medical University High-level Talent Introduction and Research Launch Fund(R2062)the Fundamental Research Funds for the Central Universities.
文摘The development and homeostasis of intestinal epithelium are mediated by actively proliferating Lgr5^(+)stem cells,which pos-sess a remarkable self-renewal and differentiation capacity.Recently,our study demonstrated that N^(6)-methyladenosine(m^(6)A)methylation was essential for the survival of colonic stem cells.Here,we show that methyltransferase-like 3(METTL3)expression is downregulated in the colon mucosa in ulcerative colitis(UC)patients and strongly associated with the differentiation and maturation of goblet cells during inflammation.In mice,depletion of Mettl3 significantly inhibits the self-renewal and differentiation of Lgr5^(+)stem cells,especially the differentiation and maturation of goblet cells,resulting in intestinal dysplasia and spontaneous inflammation.Mechanistically,Mettl3 deletion-mediated m^(6)A loss facilitates the expression levels of growth factor receptor binding protein 10(Grb10)and interferon-related developmental regulator 1(Ifrd1)via increasing their messenger RNA stability.We further demonstrate that the levels of GRB10 and IFRD1 are negatively correlated with METTL3 level in UC samples.Collectively,our data indicate that METTL3 enhances the self-renewal and differentiation of Lgr5^(+)stem cells during intestinal development and inflammation,and thus it may be a potential therapeutic target for UC treatment.
