Role of POT1 mutations in glioma initiation

POT1 突变在神经胶质瘤发生中的作用

• 批准号: 10240676
• 负责人: Ali Jalali
• 金额: $ 17.64万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-30 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10240676
• 关键词: ATRX gene; Address; Adult; Appearance; Area; Astrocytoma; Automobile Driving; Binding; Biological; Biological Models; Biology; Blood; Brain; Brain Neoplasms; Bypass; Cell Aging; Cell Proliferation; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Department chair; Development; Diffuse; Dominant-Negative Mutation; Electroporation; Embryo; Environment; Exposure to; Expression Profiling; Family; Funding; Gender; Gene Expression; Generations; Genes; Genetic; Genetic Engineering; Genetic Risk; Germ-Line Mutation; Glioma; Goals; Human; In Vitro; Incidence; Individual; Knock-out; Knowledge; Laboratories; Learning; Length; Link; MEKs; Malignant - descriptor; Malignant Neoplasms; Measures; Medicine; Mentors; Modeling; Modification; Mus; Mutation; Oncogenic; Operative Surgical Procedures; Orthologous Gene; Pathway interactions; Patients; Physicians; Population; Predisposition; Primary Brain Neoplasms; Process; Ras/Raf; Reporting; Research; Research Personnel; Risk; Role; Saliva; Scientist; Somatic Mutation; TP53 gene; Techniques; Telomerase; Testing; Therapeutic; Time; Training; Translating; Xenograft Model; career; cell transformation; cohort; college; experimental study; high risk; in utero; in vivo; in vivo Model; insight; mouse model; mutant; neurosurgery; oligodendroglioma; progenitor; programs; promoter; skills; stem cell proliferation; stem cells; symposium; telomere; tumor; tumor initiation; tumorigenesis

PROJECT SUMMARY Diffuse gliomas comprise the majority of malignant primary brain tumors and are commonly fatal. In a quest to better understand their biology, many glioma-associated mutations have been identified over the past decade. Among these, we have shown that germline mutations in POT1 are associated with increased risk of glioma development. POT1 is known to bind and protect telomeres, and several POT1 mutations in other cancers are known to result in telomere elongation. However, it is not known whether the glioma-associated POT1 mutations can cause telomere elongation, and more fundamentally, the mechanisms linking germline POT1 mutations to glioma initiation are not understood. The overall goal of this proposal is to determine how POT1 mutations facilitate glioma initiation. I hypothesize that POT1 mutations accomplish this by (1) driving cell proliferation and (2) enabling telomere elongation and bypass of replicative senescence. We plan to use glial progenitor cells (GPCs) as a model for glioma initiating cells. We will use mouse GPCs to address the first hypothesis, as mice allow in vivo modeling, and human GPCs to address the second hypothesis, as human cells are more readily capable of replicative senescence. My first Specific Aim seeks to quantify the effects of POT1 mutant expression or loss of mouse POT1 orthologs in GPC proliferation and fate in vivo, using an in utero electroporation mouse model, as well as in vitro, at various time points into adulthood. We will also assess these effects in lineage-restricted GPCs using selective promoters to drive gene expression. My second Specific Aim seeks to establish the role of POT1 mutations in replicative senescence bypass using human GPCs. We will assess the effects of POT1 mutant expression or POT1 loss on telomere length and population doubling capacity of human GPCs. We will also quantify the tumorigenesis potential of these cells in a xenograft model upon inhibition of p53 and activation of Ras pathways. These studies will help to reveal the role of POT1 mutations in glioma initiation. In the long term, I intend to leverage this knowledge to gain further insight into the mechanisms of telomere biology in glioma and translate its vulnerabilities into potential therapeutic measures. I envision a career for myself as an independent physician-scientist studying the biology and genetics of gliomas with an active brain tumor surgical practice focused on treating patients with glioma. To reach this goal, I have identified several areas of deficiency in my training that require addressing. These include knowledge of telomere biology and genetic engineering techniques as well as bench and laboratory management skills. I plan to address these deficiencies by learning from my mentor and colleagues as well as participation in courses and conferences. Baylor College of Medicine and Department of Neurosurgery provide a highly fertile environment for research, and I enjoy the full support of my mentor, advisors, and Department Chair. The program outlined in this proposal will provide me with the training to successfully complete my proposed studies as well as the track record to compete for research funding as an independent investigator.

项目摘要 弥漫性神经胶质瘤包括大多数恶性原发性脑肿瘤，并且通常是致命的。以寻求 为了更好地了解它们的生物学，在过去的十年中已经确定了许多神经胶质瘤相关的突变。 在这些研究中，我们发现POT1的生殖系突变与神经胶质瘤风险增加有关 发展已知POT1结合并保护端粒，其他癌症中的几种POT1突变是 已知导致端粒延长。然而，目前尚不清楚胶质瘤相关的POT1 突变可以导致端粒延长，更根本的是， 神经胶质瘤起始的突变尚不清楚。本提案的总体目标是确定POT1 突变促进胶质瘤起始。我假设POT1突变通过以下方式实现这一点：（1）驱动细胞 增殖和（2）使端粒延长和复制衰老的旁路。我们计划用神经胶质 祖细胞（GPC）作为胶质瘤起始细胞的模型。我们将使用鼠标GPC来解决第一个问题 第二种假设，因为小鼠允许体内建模，而人GPC解决了第二种假设，因为人 细胞更容易复制衰老。我的第一个具体目标是量化的影响， POT1突变体表达或小鼠POT1直系同源物在GPC增殖和体内命运中的损失， 子宫电穿孔小鼠模型，以及在体外，在不同的时间点进入成年期。我们还将 使用选择性启动子驱动基因表达，评估谱系限制性GPCs中的这些效应。我 第二个特定目标旨在利用以下方法确定POT 1突变在复制衰老旁路中的作用 人GPCs。我们将评估POT1突变表达或POT1缺失对端粒长度的影响， 人类GPC的人口倍增能力。我们还将量化这些细胞的肿瘤发生潜力， 在抑制p53和激活Ras途径后的异种移植模型。这些研究将有助于揭示 POT1突变在胶质瘤发生中的作用从长远来看，我打算利用这些知识， 深入了解胶质瘤中端粒生物学的机制，并将其脆弱性转化为潜在的 治疗措施。我设想自己的职业生涯是作为一个独立的物理学家和科学家， 和神经胶质瘤的遗传学与积极的脑肿瘤手术实践，重点是治疗神经胶质瘤患者。 为了达到这个目标，我已经确定了我的培训中需要解决的几个不足之处。这些 包括端粒生物学和遗传工程技术知识，以及实验室和实验室 管理能力我计划通过向我的导师和同事学习来解决这些缺陷， 参加课程和会议。贝勒医学院和神经外科系提供 一个非常肥沃的研究环境，我享受我的导师，顾问和部门的全力支持 椅子本建议书中概述的计划将为我提供成功完成我的 建议的研究以及作为独立调查员竞争研究资金的记录。