Manji Jasmine, Pipella Jasmine, Brawerman Gabriel, Thompson Peter J
Diabetes Research Envisioned and Accomplished in Manitoba (DREAM) Theme, Children's Hospital Research Institute of Manitoba, Winnipeg, MB R3E 3P4, Canada.
Department of Physiology & Pathophysiology, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, MB R3E 3P4, Canada.
Epigenomes. 2024 Mar 6;8(1):10. doi: 10.3390/epigenomes8010010.
Type 1 diabetes (T1D) is a metabolic disease resulting from progressive autoimmune destruction of insulin-producing pancreatic beta cells. Although the majority of beta cells are lost in T1D, a small subset undergoes senescence, a stress response involving growth arrest, DNA damage response, and activation of a senescence-associated secretory phenotype (SASP). SASP in beta cells of the nonobese diabetic (NOD) mouse model of T1D and primary human islets is regulated at the level of transcription by bromodomain extra-terminal (BET) proteins, but the mechanisms remain unclear. To explore how SASP is transcriptionally regulated in beta cells, we used the NOD beta cell line NIT-1 to model beta cell SASP and identified binding partners of BET protein Brd4 and explored the role of the cyclin-dependent kinase inhibitor p21. Brd4 interacted with a variety of proteins in senescent NIT-1 cells including subunits of the Ino80 chromatin remodeling complex, which was expressed in beta cells during T1D progression in NOD mice and in human beta cells of control, autoantibody-positive, and T1D donors as determined from single-cell RNA-seq data. RNAi knockdown of p21 during senescence in NIT-1 cells did not significantly impact viability or SASP. Taken together, these results suggest that Brd4 interacts with several protein partners during senescence in NIT-1 cells, some of which may play roles in SASP gene activation and that p21 is dispensable for the SASP in this beta cell model.
1型糖尿病(T1D)是一种代谢性疾病,由胰岛素生成胰腺β细胞的进行性自身免疫破坏引起。尽管在T1D中大多数β细胞会丢失,但一小部分会发生衰老,这是一种涉及生长停滞、DNA损伤反应以及衰老相关分泌表型(SASP)激活的应激反应。在T1D的非肥胖糖尿病(NOD)小鼠模型和原代人胰岛的β细胞中,SASP在转录水平上受溴结构域额外末端(BET)蛋白调控,但其机制仍不清楚。为了探究β细胞中SASP是如何进行转录调控的,我们使用NODβ细胞系NIT-1来模拟β细胞SASP,并鉴定了BET蛋白Brd4的结合伙伴,同时探究了细胞周期蛋白依赖性激酶抑制剂p21的作用。Brd4与衰老的NIT-1细胞中的多种蛋白质相互作用,包括Ino80染色质重塑复合体的亚基,根据单细胞RNA测序数据确定,该复合体在NOD小鼠T1D进展过程中的β细胞以及对照、自身抗体阳性和T1D供体的人β细胞中均有表达。在NIT-1细胞衰老过程中,通过RNA干扰敲低p21对细胞活力或SASP没有显著影响。综上所述,这些结果表明,在NIT-1细胞衰老过程中,Brd4与几种蛋白质伙伴相互作用,其中一些可能在SASP基因激活中发挥作用,并且在这个β细胞模型中,p21对于SASP是可有可无的。