The role of senescent beta cells in T1D and T2D

衰老 β 细胞在 T1D 和 T2D 中的作用

基本信息

批准号：
10708994
负责人：
KLAUS H KAESTNER
金额：
$ 72.13万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-19 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10708994
关键词：
ATAC-seq Ablation Age Alleles Animal Model Autoantibodies Autoimmunity B-Lymphocytes Backcrossings Beta Cell Biological Assay CDKN1A gene CDKN2A gene Cell Aging Cell Cycle Cell Cycle Arrest Cell physiology Cells Cellular Stress Characteristics Chronic Cre driver Cytometry DNA DNA Damage Data Development Diabetes Mellitus Disease Excision Functional disorder Gene Expression Profile Genes Genetic Genotoxic Stress Glucose Intolerance Histones Human Human Characteristics Hyperglycemia Image Inbred NOD Mice Inflammatory Insulin Insulin Resistance Insulin-Dependent Diabetes Mellitus Islet Cell Islets of Langerhans Knowledge Metabolic Metabolism Modality Modeling Molecular Morphology Mus Outcome Oxidative Stress Pancreas Pathology Pathway interactions Pharmaceutical Preparations Phenotype Population Prevalence Property Recovery Reporting Research Role Signal Transduction Stimulus TP53 gene Testing Therapeutic Transgenic Mice Tumor Suppressor Proteins Type 2 diabetic Viral Vector Work Xenograft Model Xenograft procedure acute stress age related blood glucose regulation cell injury cell type clinical practice cohort diabetic diabetic patient epigenomics gain of function genetic signature genomic tools human data improved inhibitor islet islet autoimmunity loss of function methylome mouse model non-diabetic novel prevent programs response senescence sex single-cell RNA sequencing stressor telomere tumorigenesis type I and type II diabetes type I diabetic

项目摘要

U01 Application: The role of senescent beta cells in T1D and T2D Abstract Recent studies including our own suggest a marked increase in β-cells expressing key components of cellular senescence in islets from Type 1 diabetes (T1D) and Type 2 diabetic (T2D) patients, implicating β-cell senescence as a critical contributor to islet dysfunction. Recently, it was reported that ablation of senescent cells by non-specific senolysis in mouse models of T1D and T2D improved diseease outcome. However, these studies did not determine which senescent cell type was relevant to the beneficial effect, nor did they address to what extent senescence occurs in the human endocrine pancreas before and during the development of T1D and T2D. To close this knowledge gap, in Specific Aim 1 we will determine the prevalence, transcription signatures and epigenomic landscapes of β-cell senescence in T1D, pre-T1D and T2D donors using immunostaining, imaging mass cytometry, single cell RNAseq, single cell ATACseq, DNA methylome determination and Cut-and-Tag analysis for key histone marks. In addition, we will evaluate the hypothesis that irreparable damage to telomeres drives senescence in β-cells, a possible scenario that could provide a mechanism for senescence to occur in islet cells of diabetic patients. In Specific Aim 2, we will test whether metabolic and/or inflammatory stressors drive senescence in human β-cells and determine the effect of senescence on β-cell function using scRNAseq and secretome analysis. We will evaluate if induction of senescence and the senescence-associated secretory phenotype (SASP) in human islet cells is p16 dependent, employing the pseudo-islets approach and using our hyperglycemic xeno-transplantation model to assess the direct effect of senolytics on human islet function. In Specific Aim 3, we will employ a novel transgenic mouse, the ‘SenKiller’ model, to enable cell-type specific and inducible ablation of senescent cells in any lineage including β-cells. Using this mouse model in combination with the appropriate β-cell specific Cre driver, we will provide a definitive answer to the question if senescent β-cells are critical in the development of glucose intolerance in models of T2D and islet autoimmunity in models of T1D. Together, this proposal will determine the occurrence of senescence among islet cells from T1D and T2D donors using large cohorts and multiple experimental modalities, explore the natural drivers of senescence and consequences to islet function as well as secretion of pro-inflammatory substances, and employ novel mouse models to unequivocally determine if elimination of senescent b-cells impacts diabetes progression in mouse models of T1D and T2D. The data generated here will address burning questions in the field, namely, is senescence increased in islets from diabetic patients, and are these cells important in the overall pathophysiology of T1D and T2D. These critical questions will have therapeutic relevance regarding the potential efficacy of targeting senescent β-cells with senolytic therapies.

U01 应用：衰老 β 细胞在 T1D 和 T2D 中的作用抽象的最近的研究（包括我们自己的研究）表明，表达细胞关键成分的 β 细胞显着增加。 1 型糖尿病 (T1D) 和 2 型糖尿病 (T2D) 患者的胰岛衰老，与 β 细胞有关衰老是胰岛功能障碍的关键因素，最近有报道称衰老的消融。在 T1D 和 T2D 小鼠模型中，通过非特异性衰老作用改善了细胞的疾病结果。研究没有确定哪种衰老细胞类型与有益效果相关，也没有解决在发育之前和发育过程中，人类内分泌胰腺的衰老程度如何？为了弥补这一知识差距，在具体目标 1 中，我们将确定患病率、转录情况。使用 T1D、T1D 前期和 T2D 供体中 β 细胞衰老的特征和表观基因组景观免疫染色、成像质谱流式分析、单细胞 RNAseq、单细胞 ATACseq、DNA 甲基化组关键组蛋白标记的测定和切割和标签分析此外，我们将评估以下假设。端粒不可挽回的损伤会导致β细胞衰老，这是一种可能的情况，可以提供在具体目标2中，我们将测试糖尿病患者胰岛细胞衰老的机制。代谢和/或炎症应激源驱动人类 β 细胞衰老并决定我们将使用 scRNAseq 和分泌组分析评估衰老对 β 细胞功能的影响。人类胰岛细胞的衰老和衰老相关分泌表型（SASP）是 p16 依赖性，采用伪胰岛方法并使用我们的高血糖异种移植模型评估 senolytics 对人类胰岛功能的直接影响在特定目标 3 中，我们将采用一种新颖的方法。转基因小鼠“SenKiller”模型，能够对衰老细胞进行细胞类型特异性和诱导性消融任何谱系，包括 β 细胞，将此小鼠模型与适当的 β 细胞特异性 Cre 结合使用。驱动程序，我们将为这个问题提供明确的答案：衰老的 β 细胞是否在该提案将结合 T2D 模型中的葡萄糖不耐受和 T1D 模型中的胰岛自身免疫。使用大型队列确定来自 T1D 和 T2D 供体的胰岛细胞中衰老的发生情况多种实验方式，探索衰老的自然驱动因素及其对胰岛功能的影响以及促炎物质的分泌，并采用新型小鼠模型明确地确定消除衰老 B 细胞是否会影响 T1D 和 T2D 小鼠模型的糖尿病进展。这里生成的数据将解决该领域的紧迫问题，即胰岛的衰老是否增加来自糖尿病患者，这些细胞在 T1D 和 T2D 的整体病理生理学中很重要。关于靶向衰老 β 细胞的潜在功效，关键问题将具有治疗相关性与 senolytic 疗法。