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In Vitro and In Vivo Biology of Telomere Stress Induced Senescence

端粒应激诱导衰老的体外和体内生物学

基本信息

批准号：
10663808
负责人：
Shohini Kalyani Ghosh-Choudhary
金额：
$ 5.27万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
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：
2022-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10663808
关键词：
4-Hydroxy-Tamoxifen Affect Age Aging Animal Model Aorta Apoptosis Atherosclerosis Basic Science Bioinformatics Biology CRISPR screen Cardiac Cardiac Myocytes Cardiac Output Cell Aging Cell Culture Techniques Cell Cycle Arrest Cell Death Cell Death Induction Cell Line Cell Senescence Induction Cell Survival Cell model Cells Chronic Clinical Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats DNA Damage Degenerative polyarthritis Development Diabetes Mellitus Disease Dose Limiting Drug Design Drug Targeting EFRAC Enterobacteria phage P1 Cre recombinase Estrogens Foundations Functional disorder Future Genes Genetic Predisposition to Disease Genomics Geroscience In Vitro Inflammatory Knock-out Knockout Mice Lead LoxP-flanked allele Measures Metabolic Modeling Molecular Mus Mutation Neurodegenerative Disorders Normal Cell Organ Pathology Pathway Analysis Pathway interactions Pharmaceutical Preparations Phenotype Physicians Physiological Play Population Production Proteins Reactive Oxygen Species Regulatory Pathway Research Role Scientist Stress Stroke Volume Techniques Telomere-Binding Proteins Therapeutic Therapeutic Index Thinking Thrombocytopenia Time Tissues Training Transgenic Mice Translating Translational Research age related aged bone loss cancer therapy career cell immortalization cell killing cell type chemokine cytokine deep sequencing endoplasmic reticulum stress genome-wide heart function idiopathic pulmonary fibrosis in vivo inflammatory milieu innovation knock-down knockout gene mouse model pharmacologic postmitotic postnatal response senescence side effect skills substantia spongiosa telomere tumor microenvironment whole genome

项目摘要

PROJECT SUMMARY Senescent cells have been implicated as drivers of aging and age-associated diseases. Senescent cells have sustained some form of stress that causes irreversible cell-cycle arrest, and these cells avoid apoptosis. These cells have a hyper-metabolic phenotype and secrete cytokines and chemokines resulting in the senescence associated secretory phenotype (SASP). The SASP contains multiple inflammatory components which promotes tissue dysfunction. These observations have been the impetus to develop senolytic drugs. These drugs are designed to cause the selective apoptosis of senescent cells and several senolytics have already entered clinical trials for potential use in idiopathic pulmonary fibrosis, osteoarthritis, chronic diabetes, etc. Many of these senolytic drugs have been developed via a candidate approach where drugs that were initially developed for the treatment of cancer are repurposed for the clearance of senescent cells. These drugs, unfortunately, have both on-target and off-target dose-limiting side effects such as thrombocytopenia and trabecular bone loss. Therefore, there is a need to develop senolytic drugs that target a specific underlying vulnerability of senescent cells without affecting healthy cells. In this project, we have performed an unbiased CRISPR-based, whole genome synthetic lethality screen to identify genes when knocked out, are lethal in the setting of a cell that has undergone telomere-induced senescence yet are well-tolerated when knockout in the context of a healthy, non-senescent cell. Through this screen, we identified that genetic alterations in ER stress pathways are specific vulnerabilities of senescent cells. Specifically, the targets ER-resident proteins, PARP16 and BIP, which were identified in the CRISPR screen, modulate ER stress pathways. Therefore, we hypothesize that due to their high secretory burden, senescent cells are particularly vulnerable to alterations in ER stress pathways. We propose to investigate the following: (1) Identify and characterize the set of genes which when knocked out in senescent cells cause the specific death of these cells without affecting the function of healthy cells; (2) Understand whether senescent cells that undergo telomere induced senescence have higher levels of ER stress and whether alterations in these pathways affect the viability of senescent cells and the production of SASP components; (3) Understand whether telomere-induced senescence affects the function of post- mitotic cells such as cardiomyocytes and in turn, whether cardiomyocyte senescence affect heart function. This proposal will therefore lead to a better understanding of the unique molecular vulnerabilities of senescent cells and clarify the molecular underpinnings of how cells maintain a state of senescence. Successful completion of this proposal will provide a foundation for the development of senolytics to treat a variety of aging-associated diseases. Finally, this proposal will provide me with an outstanding platform to build my basic science and clinical thinking skills so that I can pursue a career as a physician scientist.

项目总结衰老细胞被认为是衰老和衰老相关疾病的驱动因素。衰老的细胞有承受某种形式的压力，导致不可逆转的细胞周期停滞，这些细胞避免了凋亡。这些细胞具有高代谢表型，并分泌导致衰老的细胞因子和趋化因子相关分泌表型(SASP)。SASP含有多种炎性成分，促进组织功能障碍。这些观察结果一直是开发抗衰老药物的动力。这些药物被设计用来引起衰老细胞的选择性凋亡，并且已经有几种抗衰老药物进入临床试验，可能用于特发性肺纤维化、骨关节炎、慢性糖尿病等。许多这些抗衰老药物都是通过一种候选方法开发出来的，这些药物最初是为治疗癌症而开发的药物被重新用于清除衰老细胞。这些药物，不幸的是，在靶上和靶外都有限制剂量的副作用，如血小板减少和骨小梁丢失。因此，有必要开发针对特定底物的抗衰老药物。在不影响健康细胞的情况下降低衰老细胞的脆弱性。在这个项目中，我们执行了一项不偏不倚的基于CRISPR的全基因组合成致死性筛查，以识别敲除时的基因，在设置已经经历了端粒诱导的衰老但在基因敲除时仍能很好耐受的细胞健康的、不衰老的细胞的背景。通过这个筛选，我们发现内质网应激中的基因改变通路是衰老细胞特有的脆弱性。具体地说，目标是内质网驻留蛋白PARP16 和BIP，这是在CRISPR屏幕上发现的，调制内质网应激途径。因此，我们假设由于它们的高分泌负担，衰老的细胞特别容易受到 ER应激途径。我们建议进行以下研究：(1)识别和表征当在衰老细胞中被敲除导致这些细胞的特定死亡而不影响健康人的功能细胞；(2)了解经历端粒诱导衰老的衰老细胞是否有更高水平的以及这些途径的改变是否会影响衰老细胞的活性和产量 (3)了解端粒诱导的衰老是否影响细胞后功能。有丝分裂细胞如心肌细胞，进而影响心肌细胞衰老是否影响心功能。因此，这一提议将使人们更好地了解人类免疫系统的独特分子脆弱性并阐明细胞如何维持衰老状态的分子基础。这项提案的成功完成将为开发治疗癌症的感觉剂奠定基础。各种与衰老相关的疾病。最后，这项提议将为我提供一个出色的平台来构建我的基本科学和临床思维能力，使我可以追求作为一名内科科学家的职业。