m(6)A reader YTHDC1 modulates autophagy by targeting SQSTM1 in diabetic skin.
m(6)A reader YTHDC1 modulates autophagy by targeting SQSTM1 in diabetic skin.
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m6A 阅读器 YTHDC1 通过靶向糖尿病皮肤中的 SQSTM1 来调节自噬
DOI：
10.1080/15548627.2021.1974175
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发表时间：
2022-06
期刊：
影响因子：
13.3
通讯作者：
Wang, Wei
中科院分区：
文献类型：
--
作者：
Liang, Diefei;Lin, Wei-Jye;Ren, Meng;Qiu, Junxiong;Yang, Chuan;Wang, Xiaoyi;Li, Na;Zeng, Tingting;Sun, Kan;You, Lili;Yan, Li;Wang, Wei
巨噬/自噬的失调有助于糖尿病皮肤伤口愈合的延迟。已知n6 -甲基腺苷（m6A） RNA修饰在调节自噬中起关键作用。本研究发现，在两种短期高糖治疗的人角质细胞细胞系中，以及糖尿病患者和长期高血糖的db/db小鼠模型的表皮中，自噬受体SQSTM1/p62 （sequestosome 1）显著下调。敲低SQSTM1导致自噬通量受损，这与高糖处理在角质形成细胞中的结果一致。此外，与SQSTM1 mRNA相互作用的m6A读取器蛋白YTHDC1（含YTH结构域1）在急性和慢性高血糖作用下，在角质形成细胞中均下调。YTHDC1的下调会影响角质形成细胞的生物学功能，包括凋亡率增加和伤口愈合能力受损。此外，内源性YTHDC1的敲低导致角化细胞自噬通量的阻断，而YTHDC1的过表达挽救了高糖诱导的自噬通量的阻断。在体内，内源性Ythdc1或Sqstm1的下调抑制了表皮的自噬，延迟了伤口愈合。有趣的是，我们发现YTHDC1的降低驱动细胞核中SQSTM1 mRNA的降解。此外，YTHDC1与ELAVL1/HuR （ELAV样RNA结合蛋白1）相互作用并协同调节SQSTM1的表达。总的来说，本研究揭示了YTHDC1在糖尿病角化细胞中通过调节SQSTM1核mRNA的稳定性来调节自噬的先前未被认识的功能。缩写：ACTB：肌动蛋白；AGEs：糖基化终产物；艾尔:自吞噬泡;美联社:自噬小体;ATG：自噬相关；AKT: AKT丝氨酸/苏氨酸激酶；ANOVA：方差分析；BECN1: beclin 1；Co-IP: co-immunoprecipitation;DEGs：差异表达基因；DM：糖尿病；ELAVL1: ELAV样RNA结合蛋白1；FTO: α -酮戊二酸依赖双加氧酶；G:葡萄糖;HaCaT：人角质细胞；GO：基因本体；GSEA：基因集富集分析；他:hematoxylin-eosin;包含IHC:免疫组织化学;IRS：免疫反应评分；KEAP1: kelch样ECH相关蛋白1；京都基因与基因组百科全书；m6A: N6-methyladenosine;M:甘露醇;MANOVA：多变量方差分析；MAP1LC3：微管相关蛋白1轻链3；MAP1LC3B：微管相关蛋白1轻链3 β；MeRIP：甲基化RNA免疫沉淀；METTL3：甲基转移酶3，n6 -腺苷-甲基转移酶络合物催化亚基；MTOR：雷帕霉素激酶的机制靶点MTORC1：雷帕霉素复合物1的机制靶点NBR1: NBR1自噬货物受体；NFE2L2：核因子，红细胞2样2；NG：正常葡萄糖；NHEK：正常人上皮角质细胞；OE: overexpressing;p -:含磷的;PI：碘化丙啶；蛋白-蛋白相互作用网络；rbp: RNA结合蛋白；RIP: RNA免疫沉淀；RNA-seq: rna序列;RNU6-1: RNA， U6小核1；ROS：活性氧；sirna：小干扰rna；SQSTM1: sequestosome 1；SRSF：富含丝氨酸和精氨酸的剪接因子；T2DM: 2型糖尿病；TEM：透射电子显微镜；TUBB: I类微管蛋白；WT:野生型;YTHDC1：包含1的YTH域。
Dysregulation of macroautophagy/autophagy contributes to the delay of wound healing in diabetic skin. N6-methyladenosine (m6A) RNA modification is known to play a critical role in regulating autophagy. In this study, it was found that SQSTM1/p62 (sequestosome 1), an autophagy receptor, was significantly downregulated in two human keratinocyte cells lines with short-term high-glucose treatment, as well as in the epidermis of diabetic patients and a db/db mouse model with long-term hyperglycemia. Knockdown of SQSTM1 led to the impairment of autophagic flux, which was consistent with the results of high-glucose treatment in keratinocytes. Moreover, the m6A reader protein YTHDC1 (YTH domain containing 1), which interacted with SQSTM1 mRNA, was downregulated in keratinocytes under both the acute and chronic effects of hyperglycemia. Knockdown of YTHDC1 affected biological functions of keratinocytes, which included increased apoptosis rates and impaired wound-healing capacity. In addition, knockdown of endogenous YTHDC1 resulted in a blockade of autophagic flux in keratinocytes, while overexpression of YTHDC1 rescued the blockade of autophagic flux induced by high glucose. In vivo, knockdown of endogenous Ythdc1 or Sqstm1 inhibited autophagy in the epidermis and delayed wound healing. Interestingly, we found that a decrease of YTHDC1 drove SQSTM1 mRNA degradation in the nucleus. Furthermore, the results revealed that YTHDC1 interacted and cooperated with ELAVL1/HuR (ELAV like RNA binding protein 1) in modulating the expression of SQSTM1. Collectively, this study uncovered a previously unrecognized function for YTHDC1 in modulating autophagy via regulating the stability of SQSTM1 nuclear mRNA in diabetic keratinocytes. Abbreviations: ACTB: actin beta; AGEs: glycation end products; AL: autolysosome; AP: autophagosome; ATG: autophagy related; AKT: AKT serine/threonine kinase; ANOVA: analysis of variance; BECN1: beclin 1; Co-IP: co-immunoprecipitation; DEGs: differentially expressed genes; DM: diabetes mellitus; ELAVL1: ELAV like RNA binding protein 1; FTO: FTO alpha-ketoglutarate dependent dioxygenase; G: glucose; HaCaT: human keratinocyte; GO: Gene Ontology; GSEA: Gene Set Enrichment Analysis; HE: hematoxylin-eosin; IHC: immunohistochemical; IRS: immunoreactive score; KEAP1: kelch like ECH associated protein 1; KEGG: Kyoto Encyclopedia of Genes and Genomes; m6A: N6-methyladenosine; M: mannitol; MANOVA: multivariate analysis of variance; MAP1LC3: microtubule associated protein 1 light chain 3; MAP1LC3B: microtubule associated protein 1 light chain 3 beta; MeRIP: methylated RNA immunoprecipitation; METTL3: methyltransferase 3, N6-adenosine-methytransferase complex catalytic subunit; MTOR: mechanistic target of rapamycin kinase; MTORC1: mechanistic target of rapamycin complex 1; NBR1: NBR1 autophagy cargo receptor; NFE2L2: nuclear factor, erythroid 2 like 2; NG: normal glucose; NHEK: normal human epithelial keratinocyte; OE: overexpressing; p-: phospho-; PI: propidium iodide; PPIN: Protein-Protein Interaction Network; RBPs: RNA binding proteins; RIP: RNA immunoprecipitation; RNA-seq: RNA-sequence; RNU6–1: RNA, U6 small nuclear 1; ROS: reactive oxygen species; siRNAs: small interfering RNAs; SQSTM1: sequestosome 1; SRSF: serine and arginine rich splicing factor; T2DM: type 2 diabetes mellitus; TEM: transmission electron microscopy; TUBB: tubulin beta class I; WT: wild-type; YTHDC1: YTH domain containing 1.
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影响因子：
4.8
作者：
Jain, Ashish;Lamark, Trond;Johansen, Terje
通讯作者：
Johansen, Terje
DOI：
10.1074/jbc.m109.096545
发表时间：
2010-05-28
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Copple IM;Lister A;Obeng AD;Kitteringham NR;Jenkins RE;Layfield R;Foster BJ;Goldring CE;Park BK
通讯作者：
Park BK
p62/SQSTM1形成自噬降解的蛋白质聚集体，并对亨廷顿蛋白诱导的细胞死亡具有保护作用。
影响因子：
7.8
作者：
Bjorkoy, Geir;Lamark, Trond;Brech, Andreas;Outzen, Heidi;Perander, Maria;Overvatn, Aud;Stenmark, Harald;Johansen, Terje
通讯作者：
Johansen, Terje
影响因子：
3.6
作者：
Kim, Jae-Hong;Yoon, Na Young;Choi, Eung Ho
通讯作者：
Choi, Eung Ho
影响因子：
13.3
作者：
Haidar, Mansour;Asselbergh, Bob;Timmerman, Vincent
通讯作者：
Timmerman, Vincent