DNA damage to β cells in culture recapitulates features of senescent β cells that accumulate in type 1 diabetes.
DNA damage to β cells in culture recapitulates features of senescent β cells that accumulate in type 1 diabetes.
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DOI：
10.1016/j.molmet.2022.101524
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发表时间：
2022-08
影响因子：
8.1
通讯作者：
Thompson, Peter J.
中科院分区：
文献类型：
--
作者：
Brawerman, Gabriel;Pipella, Jasmine;Thompson, Peter J.
1型糖尿病(T1D)的特征是自身免疫破坏导致产生胰岛素的胰腺β细胞进行性丧失。除了β细胞死亡，最近的研究表明，β细胞亚群在T1D期间获得功能障碍。我们以前曾报道，在T1D的发病过程中，经历DNA损伤反应和衰老的β细胞会积累。然而，β细胞的衰老如何发展的问题还没有被研究过。在这里，我们使用培养模型验证了这一假设，即在遗传易感性的背景下未修复的DNA损伤会触发β细胞衰老，培养模型包括来自T1D敏感的非肥胖型糖尿病(NOD)株的小鼠β细胞系、人类供体胰岛和Endocβ细胞。用足叶乙甙或博莱霉素化学诱导细胞或胰岛DNA损伤，结合免疫印迹、qRT-PCR、Luminex分析、流式细胞术和组织化学染色等方法分析细胞或胰岛的衰老表型。RNA-SEQ被用来描述经历DDR和衰老的人类胰岛的整体转录变化。用胰岛素ELISA定量测定化学诱导衰老的人胰岛、Endocβ细胞和小鼠β细胞系培养中葡萄糖刺激的胰岛素分泌。NIT1细胞的亚致死性DNA损伤导致了几个典型的衰老特征，包括持续的DDR激活，生长停滞，形态变大，扁平和衰老相关的分泌表型，类似于NOD小鼠原代β细胞在T1D期间发生的情况。这些表型在NIT1细胞和来自非T1D敏感小鼠品系的MIN6β细胞系之间存在差异。对来自两个不同捐赠者的人类胰岛DNA损伤诱导的衰老的RNA-SEQ分析显示，P53转录程序和存活率和SASP基因上调，这种反应在捐赠者之间存在差异。在人类胰岛中，供体间的差异也在蛋白水平上的持续DDR激活和SASP的程度上明显。值得注意的是，化学诱导的DNA损伤也导致了endo-βH5人β细胞的DDR激活和衰老表型，证实这种反应可以直接发生在人β细胞系中。最后，与人胰岛和Endocβ细胞相比，DNA损伤导致小鼠β细胞系中葡萄糖刺激的胰岛素分泌的不同影响。综上所述，这些发现表明，在NOD小鼠和人类T1D发育过程中积累的衰老β细胞的一些表型可以通过化学诱导培养中的小鼠β细胞系、人胰岛和Endocβ细胞的DNA损伤来模拟。来自不同小鼠品系和不同人类胰岛供体的β细胞和Endocβ细胞之间的差异突出了物种差异和遗传背景在改变β细胞对DNA损伤的反应及其对胰岛素分泌的影响方面的作用。这些培养模型将为理解β细胞在T1D衰老的某些机制提供有用的工具。DNA损伤可引起小鼠β细胞系衰老表型。DNA损伤诱导人类胰岛和EnDoc-βH5细胞的P53程序和衰老表型。DNA损伤诱导的衰老导致不同的培养模式对胰岛素分泌的不同影响。
Type 1 Diabetes (T1D) is characterized by progressive loss of insulin-producing pancreatic β cells as a result of autoimmune destruction. In addition to β cell death, recent work has shown that subpopulations of β cells acquire dysfunction during T1D. We previously reported that β cells undergoing a DNA damage response (DDR) and senescence accumulate during the pathogenesis of T1D. However, the question of how senescence develops in β cells has not been investigated. Here, we tested the hypothesis that unrepaired DNA damage in the context of genetic susceptibility triggers β cell senescence using culture models including the mouse NIT1 β cell line derived from the T1D-susceptible nonobese diabetic (NOD) strain, human donor islets and EndoC β cells. DNA damage was chemically induced using etoposide or bleomycin and cells or islets were analyzed by a combination of molecular assays for senescence phenotypes including Western blotting, qRT-PCR, Luminex assays, flow cytometry and histochemical staining. RNA-seq was carried out to profile global transcriptomic changes in human islets undergoing DDR and senescence. Insulin ELISAs were used to quantify glucose-stimulated insulin secretion from chemically-induced senescent human islets, EndoC β cells and mouse β cell lines in culture. Sub-lethal DNA damage in NIT1 cells led to several classical hallmarks of senescence including sustained DDR activation, growth arrest, enlarged flattened morphology and a senescence-associated secretory phenotype (SASP) resembling what occurs in primary β cells during T1D in NOD mice. These phenotypes differed between NIT1 cells and the MIN6 β cell line derived from a non-T1D susceptible mouse strain. RNA-seq analysis of DNA damage-induced senescence in human islets from two different donors revealed a p53 transcriptional program and upregulation of prosurvival and SASP genes, with inter-donor variability in this response. Inter-donor variability in human islets was also apparent in the extent of persistent DDR activation and SASP at the protein level. Notably, chemically induced DNA damage also led to DDR activation and senescent phenotypes in EndoC-βH5 human β cells, confirming that this response can occur directly in a human β cell line. Finally, DNA damage led to different effects on glucose-stimulated insulin secretion in mouse β cell lines as compared with human islets and EndoC β cells. Taken together, these findings suggest that some of the phenotypes of senescent β cells that accumulate during the development of T1D in the NOD mouse and humans can be modeled by chemically induced DNA damage to mouse β cell lines, human islets and EndoC β cells in culture. The differences between β cells from different mouse strains and different human islet donors and EndoC β cells highlights species differences and the role for genetic background in modifying the β cell response to DNA damage and its effects on insulin secretion. These culture models will be useful tools to understand some of the mechanisms of β cell senescence in T1D. DNA damage elicits senescent phenotypes in mouse β cell lines. DNA damage induces a p53 program and senescent phenotypes in human islets and EndoC-βH5 cells. DNA damage-induced senescence leads to different effects on insulin secretion depending on the culture model.
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影响因子：
30.8
作者：
Dooley J;Tian L;Schonefeldt S;Delghingaro-Augusto V;Garcia-Perez JE;Pasciuto E;Di Marino D;Carr EJ;Oskolkov N;Lyssenko V;Franckaert D;Lagou V;Overbergh L;Vandenbussche J;Allemeersch J;Chabot-Roy G;Dahlstrom JE;Laybutt DR;Petrovsky N;Socha L;Gevaert K;Jetten AM;Lambrechts D;Linterman MA;Goodnow CC;Nolan CJ;Lesage S;Schlenner SM;Liston A
通讯作者：
Liston A
影响因子：
3.3
作者：
Freund A;Laberge RM;Demaria M;Campisi J
通讯作者：
Campisi J
伊马替尼治疗新发 1 型糖尿病患者：一项多中心、随机、双盲、安慰剂对照 2 期试验。
DOI：
10.1016/s2213-8587(21)00139-x
发表时间：
2021-08
期刊：
The lancet. Diabetes & endocrinology
影响因子：
--
作者：
Gitelman SE;Bundy BN;Ferrannini E;Lim N;Blanchfield JL;DiMeglio LA;Felner EI;Gaglia JL;Gottlieb PA;Long SA;Mari A;Mirmira RG;Raskin P;Sanda S;Tsalikian E;Wentworth JM;Willi SM;Krischer JP;Bluestone JA;Gleevec Trial Study Group
通讯作者：
Gleevec Trial Study Group
影响因子：
8
作者：
Fischer M
通讯作者：
Fischer M
影响因子：
8.1
作者：
Benazra, Marion;Lecomte, Marie-Jose;Ravassard, Philippe
通讯作者：
Ravassard, Philippe