The role of senescent beta cells in T1D and T2D

衰老 β 细胞在 T1D 和 T2D 中的作用

• 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
• 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

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：Senscent Beta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）患者的胰岛感应 感应是胰岛功能障碍的关键因素。最近，据报道，感应的消融 在T1D和T2D的小鼠模型中，通过非特异性鼻溶细胞改善了疾病结果。但是，这些 研究并未确定哪种感觉细胞类型与有益效应相关，也没有解决 在开发之前和期间，人类内分泌胰腺发生了多大程度的感受 T1D和T2D。要缩小这一知识差距，在特定目标1中，我们将确定转录的患病率 使用T1D，T1D和T2D供体中β细胞感应的特征和表观基因组景观 免疫染色，成像质量细胞术，单细胞RNASEQ，单细胞Atacseq，DNA甲基团 关键组蛋白标记的确定和剪切分析。此外，我们将评估以下假设 无法弥补的对端粒的损害驱动β细胞衰老，这种情况可能会提供 在糖尿病患者的胰岛细胞中发生感应的机制。在特定目标2中，我们将测试是否 代谢和/或炎性应激源在人β细胞中驱动感应，并确定 使用SCRNASEQ和分泌组分析对β细胞功能的感应。我们将评估是否归纳 人类胰岛细胞中的感应和与感应相关的秘书表型（SASP）是p16 依赖，采用伪islet方法，并使用我们的高血糖异种移植模型 评估鼻溶剂对人类胰岛功能的直接影响。在特定目标3中，我们将采用小说 转基因小鼠是“ senkiller”模型，以实现细胞类型的特异性并诱导的感觉细胞中的消融 包括β细胞在内的任何血统。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，我们将为一个问题提供一个确定的答案，即 T1D模型中T2D和Islet自身免疫模型中的葡萄糖intlerance。在一起，这个建议将 确定使用大型队列和 多种实验方式，探索感应的自然驱动因素和胰岛功能的后果 以及促炎物质的分泌，以及员工新颖的小鼠模型，以明确地进行 确定消除感觉B细胞是否影响T1D和T2D小鼠模型中的糖尿病进展。 此处生成的数据将解决现场的燃烧问题，即胰岛中的敏感性增加 来自糖尿病患者，这些细胞在T1D和T2D的总体病理生理中很重要。这些 关键问题将具有有关靶向senscentβ细胞的潜在有效性的热相关性 塞溶剂疗法。