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自噬通过 NFE2L2 和 DDIT3 介导的机制调控 GH-IGF1 系统来控制新生儿肌发生。

Autophagy controls neonatal myogenesis by regulating the GH-IGF1 system through a NFE2L2- and DDIT3-mediated mechanism.

机构信息

a Unit of Clinical Pharmacology , University Hospital "Luigi Sacco"-ASST Fatebenefratelli Sacco , Milano , Italy.

b Department of Biomedical and Clinical Sciences "Luigi Sacco" , Università degli Studi di Milano , Milano , Italy.

出版信息

Autophagy. 2019 Jan;15(1):58-77. doi: 10.1080/15548627.2018.1507439. Epub 2018 Sep 10.

Abstract

Macroautophagy/autophagy is emerging as an important process in adult muscle stem cells functions: it regulates metabolic reprogramming during activation from a quiescent state, maintains stemness and prevents senescence. We now show that autophagy is specifically required for neonatal myogenesis and muscle development. Specific deletion of Atg7 in PAX7 (paired box 7) precursors led in mice to a dwarf phenotype, with an effect restricted to the neonatal phase of muscle development. Atg7 knockdown suppressed neonatal satellite cell (nSC) proliferation and differentiation, downregulating the GH-IGF1 functions. When we disrupted autophagy, NFE2L2/NRF2 (nuclear factor, erythroid 2 like 2) accumulated in muscle and nSCs and negatively modulated DDIT3/CHOP (DNA-damage inducible transcript 3) expression. Lower levels of DDIT3 were responsible for reduced GHR expression leading to impaired local production of IGF1. Our results conclusively identify a novel autophagy-dependent pathway that regulates nSC behavior and indicate that autophagy is required for skeletal muscle development in the neonatal phase. Abbreviations: AKT/protein kinase B: Thymoma viral proto-oncogene; ASCs: adult stem cells; ATF4: activating transcription factor 4; ATG7: autophagy related 7; BAT: brown adipose tissue; BMP: bone morphogenetic protein; CEBPB: CCAAT/enhancer binding protein (C/EBP), beta; CSA: cross sectional area; CTNNB1: catenin (cadherin associated protein), beta 1; DDIT3: DNA-damage inducible transcript 3; DM: differentiation medium; E: embryonic stage; EIF2AK3/PERK; EIF4EBP1: eukaryotic translation initiation factor 2 alpha kinase 3; eukaryotic translation initiation factor 4E binding protein 1; ER: endoplasmic reticulum; FGF21: fibroblast growth factor 21; GH: growth hormone; GHR: growth hormone receptor; HSCs: hematopoietic stem cells; IGF1: insulin-like growth factor 1; ITGAM: integrin alpha M; KEAP1: kelch-like ECH-associated protein 1; LY6A/Sca-1; MAP1LC3: lymphocyte antigen 6 complex, locus A; microtubule-associated protein 1 light chain 3; MAPK1/ERK2: mitogen-activated protein kinase 1; MAPK3/ERK1: mitogen-activated protein kinase 3; miRNAs: microRNAs; MSCs: mesenchymal stem cells; MTOR: mechanistic target of rapamycin kinase; mtUPR: mitochondrial unfolded protein response; MYF5: myogenic factor 5; MYH: myosin, heavy polypeptide; MYOD1: myogenic differentiation 1; MYOG: myogenin; NFE2L2: nuclear factor, erythroid derived 2, like 2; nSC: neonatal satellite cells; NSCs: neuronal stem cells; P: postnatal day; PAX7: paired box 7; PECAM1: platelet/endothelial cell adhesion molecule 1; PPARG: peroxisome proliferator activated receptor gamma; PTPRC: protein tyrosine phosphatase, receptor type, C; ROS: reactive oxygen species; RPS6: ribosomal protein S6; SCs: adult satellite cells; SQSTM1: sequestosome 1; STAT5: signal transducer and activator of transcription 5; TGFB1: transforming growth factor beta 1; WAT: white adipose tissue; WT: wild type.

摘要

自噬作为成体肌肉干细胞功能的一个重要过程正在兴起

它调节从静止状态激活过程中的代谢重编程,维持干细胞特性并防止衰老。我们现在表明,自噬对于新生儿肌肉发生和肌肉发育是特异性所必需的。特异性敲除 PAX7(配对盒 7)前体细胞中的 Atg7 会导致小鼠出现矮小表型,其作用仅限于肌肉发育的新生儿期。Atg7 敲低抑制了新生儿卫星细胞(nSC)的增殖和分化,下调了 GH-IGF1 功能。当我们破坏自噬时,NFE2L2/NRF2(核因子,红细胞 2 样 2)在肌肉和 nSCs 中积累,并负调控 DDIT3/CHOP(DNA 损伤诱导转录 3)的表达。DDIT3 水平降低导致 GHR 表达减少,从而导致局部 IGF1 产生受损。我们的结果明确确定了一种新的自噬依赖性途径,该途径调节 nSC 行为,并表明自噬是新生儿期骨骼肌发育所必需的。缩写:AKT/protein kinase B:胸腺瘤病毒原癌基因;ASCs:成体干细胞;ATF4:激活转录因子 4;ATG7:自噬相关 7;BAT:棕色脂肪组织;BMP:骨形态发生蛋白;CEBPB:CCAAT/enhancer binding protein(C/EBP),beta;CSA:横截面积;CTNNB1:连接蛋白(钙粘蛋白相关蛋白),beta 1;DDIT3:DNA 损伤诱导转录 3;DM:分化培养基;E:胚胎阶段;EIF2AK3/PERK;EIF4EBP1:真核翻译起始因子 2 alpha 激酶 3;真核翻译起始因子 4E 结合蛋白 1;ER:内质网;FGF21:成纤维细胞生长因子 21;GH:生长激素;GHR:生长激素受体;HSCs:造血干细胞;IGF1:胰岛素样生长因子 1;ITGAM:整合素 alpha M;KEAP1:kelch-like ECH-associated protein 1;LY6A/Sca-1;MAP1LC3:淋巴细胞抗原 6 复合物,座 A;微管相关蛋白 1 轻链 3;MAPK1/ERK2:丝裂原活化蛋白激酶 1;MAPK3/ERK1:丝裂原活化蛋白激酶 3;miRNAs:microRNAs;MSCs:间充质干细胞;MTOR:雷帕霉素靶蛋白激酶;mtUPR:线粒体未折叠蛋白反应;MYF5:成肌因子 5;MYH:肌球蛋白,重链多肽;MYOD1:成肌分化 1;MYOG:肌生成素;NFE2L2:红细胞衍生 2 样,核因子 2;nSC:新生儿卫星细胞;NSCs:神经干细胞;P:出生后第天;PAX7:配对盒 7;PECAM1:血小板/内皮细胞粘附分子 1;PPARG:过氧化物酶体增殖物激活受体 gamma;PTPRC:蛋白酪氨酸磷酸酶,受体型,C;ROS:活性氧;RPS6:核糖体蛋白 S6;SCs:成年卫星细胞;SQSTM1:自噬体相关蛋白 1;STAT5:信号转导和转录激活因子 5;TGFB1:转化生长因子 beta 1;WAT:白色脂肪组织;WT:野生型。

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