Role of POT1 mutations in glioma initiation

POT1 突变在神经胶质瘤发生中的作用

基本信息

批准号：
10022347
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
项目类别：
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/10022347
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

项目摘要

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.

项目概要弥漫性神经胶质瘤占大多数恶性原发性脑肿瘤，通常是致命的。为了寻求为了更好地了解其生物学特性，在过去十年中已经发现了许多与神经胶质瘤相关的突变。其中，我们发现 POT1 种系突变与神经胶质瘤风险增加相关发展。已知 POT1 可以结合和保护端粒，其他癌症中的几种 POT1 突变已知会导致端粒延长。然而，目前尚不清楚是否与神经胶质瘤相关的 POT1 突变会导致端粒延长，更根本的是，连接种系 POT1 的机制神经胶质瘤起始的突变尚不清楚。该提案的总体目标是确定 POT1 如何突变促进神经胶质瘤的发生。我假设 POT1 突变通过 (1) 驱动细胞来实现这一点增殖和（2）使端粒延长并绕过复制衰老。我们计划使用胶质祖细胞（GPC）作为神经胶质瘤起始细胞的模型。我们将使用鼠标 GPC 来解决第一个问题假设，因为小鼠允许进行体内建模，而人类 GPC 则解决了第二个假设，因为人类细胞更容易复制衰老。我的第一个具体目标旨在量化以下因素的影响 POT1 突变体表达或小鼠 POT1 直向同源物缺失对 GPC 增殖和体内命运的影响，使用 in 子宫电穿孔小鼠模型以及体外，在成年期的各个时间点。我们也会使用选择性启动子驱动基因表达来评估谱系限制性 GPC 中的这些影响。我的第二个具体目标旨在利用以下方法确定 POT1 突变在复制衰老旁路中的作用人类 GPC。我们将评估 POT1 突变体表达或 POT1 丢失对端粒长度和人类 GPC 的群体倍增能力。我们还将量化这些细胞的肿瘤发生潜力抑制 p53 和激活 Ras 途径的异种移植模型。这些研究将有助于揭示 POT1 突变在神经胶质瘤发生中的作用。从长远来看，我打算利用这些知识来获得更多深入了解神经胶质瘤端粒生物学机制，并将其脆弱性转化为潜力治疗措施。我设想自己的职业生涯是作为一名研究生物学的独立医师科学家和神经胶质瘤的遗传学，并积极开展脑肿瘤外科手术，重点治疗神经胶质瘤患者。为了实现这一目标，我发现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基因工程技术以及实验台和实验室的知识管理技能。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提供一个非常丰富的研究环境，我享受导师、顾问和部门的全力支持椅子。本提案中概述的计划将为我提供成功完成我的培训提出的研究以及作为独立研究者竞争研究经费的记录。