THE ROLE OF TELOMERASE REGULATORS IN TELOMERE MAINTENANCE AND GENOMIC INSTABILITY

端粒酶调节剂在端粒维持和基因组不稳定中的作用

基本信息

批准号：
9215460
负责人：
Zhou Songyang
金额：
$ 36.26万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-01-01 至 2021-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9215460
关键词：
Adopted Affect Animal Model Biogenesis Biological Assay Biology CRISPR/Cas technology Cancer Cell Growth Cancer Etiology Cell Cycle Cell model Cells Chronic Lymphocytic Leukemia Complement Complex DNA DNA Damage DNA Folding DNA biosynthesis Data Development Dyskeratosis Congenita Fluorescence Functional disorder Genomic Instability Grant Growth Holoenzymes Human Investigation Knock-out Lead Length Libraries Link Malignant Neoplasms Mammalian Chromosomes Mediating Methods Molecular Molecular Conformation Mus Mutation Names Oncogenic Open Reading Frames Pathway interactions Patients Physiological Predisposition Proteins Regulation Role Study models Telomerase Telomere Capping Telomere Maintenance Telomere Shortening Telomere-Binding Proteins Validation Vitronectin Work Xenograft Model base cancer cell cancer diagnosis cancer therapy effective therapy follow-up genetic regulatory protein genome-wide analysis melanoma mutant new therapeutic target novel overexpression prevent promoter screening telomere telomere loss therapeutic target trafficking treatment strategy tumor tumor growth whole genome

项目摘要

Project Summary Mammalian chromosome ends or telomeres are tightly regulated by the telomerase that mediates telomere elongation and telomere-binding proteins that cap and protect telomere ends. Telomere DNA normally adopts a closed conformation, capped and protected by a multitude of telomere-binding proteins, to prevent DNA damage and genome instability. Telomeres become open and linear during DNA replication to enable telomerase access for telomere elongation. Exposed and critically short telomeres, as a result of mutations in telomerase and telomere regulators, also become open and susceptible to damage and genome instability, ultimately leading to cancer. Mutations in telomere-binding proteins and the TERT promoter have been identified in a number of cancers. Most cancer cells have up-regulated telomerase expression and activities, and cancer cells appear highly sensitive to perturbations in telomerase activities and telomere capping, making the telomerase attractive for therapeutic targeting. A comprehensive study of telomerase regulators therefore should greatly facilitate our understanding of telomerase dysregulation in cancer and the discovery of new drug targets. We have developed an arrayed whole-genome protein interaction network screening strategy based on the Bi-molecular Fluorescence Protein Complementation (BiFC) assay. A pilot TERT BiFC screen identified several proteins as key components of the telomerase complex, including a protein we named TARP1 that has never been characterized before. We propose here to screen genome wide for cell cycle-dependent regulators of the telomerase, and to examine the mechanisms and function of the TARP1-telomerase complex. We will use inducible TARP1 knockout cells generated by CRISPR/Cas9 as well as mouse xenograft models for these studies. Our work will have important implications in devising effective treatment strategies for cancers that result from telomere dysfunction induced genome instability.

项目摘要哺乳动物染色体末端或端粒受介导的端粒酶严格调节端粒伸长率和端粒结合蛋白，这些蛋白限制和保护端粒末端。端粒DNA 通常采用封闭的构型，受到多种端粒结合蛋白的限制和保护防止DNA损伤和基因组不稳定性。在DNA复制期间，端粒变得开放和线性启用端粒酶访问端粒伸长率。由于端粒酶和端粒调节剂的突变也变得开放并容易受到损害和基因组不稳定，最终导致癌症。端粒结合蛋白和TERT启动子的突变具有在许多癌症中被鉴定出来。大多数癌细胞具有上调的端粒酶表达和活动，癌细胞对端粒酶和端粒的扰动显着敏感封盖，使端粒酶对治疗靶向具有吸引力。端粒酶的全面研究因此，监管机构应极大地促进我们对癌症和端粒酶失调的理解发现新药靶标。我们已经开发了一个数组的全基因组蛋白质相互作用网络基于双分子荧光蛋白互补（BIFC）测定法的筛选策略。飞行员 TERT BIFC屏幕将几种蛋白质确定为端粒酶复合物的关键成分，包括A 我们命名为Tarp1的蛋白质以前从未有过特征。我们在这里提议筛选基因组宽用于端粒酶的细胞周期依赖性调节剂，并检查 TARP1-Telomerase复合物。我们还将使用CRISPR/CAS9产生的诱导TARP1敲除单元作为这些研究的小鼠异种移植模型。我们的工作将在设计有效的情况下具有重要意义端粒功能障碍引起的癌症的治疗策略诱导基因组不稳定性。