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.

项目摘要衰老细胞被认为是衰老和年龄相关疾病的驱动因素。衰老细胞具有持续某种形式的应力会导致不可逆的细胞周期停滞，这些细胞避免了凋亡。这些细胞具有超级代谢表型，分泌细胞因子和趋化因子，导致衰老相关的分泌表型（SASP）。 SASP包含多种炎症成分，这些成分促进组织功能障碍。这些观察结果一直是开发鼻溶剂的动力。这些药物旨在引起衰老细胞的选择性凋亡，并且已经有几种鼻溶剂进入特发性肺纤维化，骨关节炎，慢性糖尿病等的临床试验。这些鼻溶剂中的许多是通过候选方法开发的，最初是药物为癌症治疗而开发的用于清除衰老细胞。这些药物，不幸的是，具有靶标和脱靶剂量限制副作用，例如血小板减少症和小梁骨质流失。因此，有必要开发针对特定基础的鼻溶剂衰老细胞的脆弱性而不会影响健康的细胞。在这个项目中，我们进行了公正基于CRISPR的全基因组合成杀伤力筛查以识别被击倒时鉴定基因，在在敲除时，经历了端粒引起的衰老的细胞的设置却得到了良好的耐受性。健康，非元素细胞的背景。通过此屏幕，我们确定了ER应力的遗传改变途径是衰老细胞的特定漏洞。具体而言，靶标ER居民蛋白，PARP16 在CRISPR屏幕上鉴定出的BIP调节ER应力途径。因此，我们假设由于其高分泌负担，衰老细胞特别容易受到改变 ER应力途径。我们建议调查以下情况：（1）识别和表征一组基因在衰老细胞中被撞倒会导致这些细胞的特定死亡，而不会影响健康的功能细胞；（2）了解经历端粒诱导衰老的衰老细胞是否具有更高的水平 ER应力以及这些途径的改变是否影响衰老细胞的生存能力和生产 SASP组件；（3）了解端粒诱导的衰老是否影响 - 有丝分裂细胞（例如心肌细胞）以及心肌细胞衰老是否会影响心脏功能。因此，该建议将使人们更好地了解衰老细胞并阐明细胞如何保持衰老状态的分子基础。该提案的成功完成将为开发Senolotics的发展提供基础各种衰老相关疾病。最后，该建议将为我提供一个出色的平台我的基础科学和临床思维技巧，以便我可以从事医师科学家的职业。