Understanding the divergent functions of mutant p53

项目摘要/摘要 肿瘤抑制基因TP53的突变是癌症中最常见的基因改变，超过100 在TP53中发现了明显的和反复出现的错义突变。到目前为止几乎所有研究过的p53突变体 已经失去了与DNA结合的能力，从而削弱了它作为转录因子的功能，而且似乎很可能 这种分子功能在很大程度上解释了它在肿瘤形成中的作用。此外，许多研究都有 发现单个突变体的功能获得或致癌特性，其范围超出野生型丧失的范围 功能，最显著的是促进侵袭和转移的能力。然而，哪一种特定的TP53 突变导致肿瘤的不同发展及其分子机制 人们对此仍然知之甚少。鉴于P53是一种转录因子，而且癌症的很大一部分- 相关突变，包括C132Y，发生在DNA结合域，我假设p53突变 通过深刻改变细胞转录组促进分化肿瘤的形成和进展 突变体特有的方式。 目的1：研究突变型p53基因C132Y的转移潜能。在初步研究中，我确定了 癌症相关突变，C132Y，作为一种比其他突变具有更大转移潜力的等位基因。在这 目的：对该突变体进行体外和体内功能鉴定，目的是确定：1) 突变体是否对转移是必需的，以及2)突变体促进转移的分子机制 转移。 目的2：p53错义突变体的系统特征。许多研究，包括我们自己的一些研究 实验室提供的数据表明，独特的错义突变体P53的功能可能存在差异 蛋白质。这些功能是什么，它们在突变体之间的保守程度仍然很差 明白了。为了解决这个问题，我建议：1)对细胞转录组进行系统的描述 含有不同的突变型p53蛋白和2)每个突变型的前两个突变型的详细特征 转录簇以探索突变体发挥作用的机制。 培训计划：申请人将与一个由导师和合作者组成的跨学科团队合作，以获得 在细胞和分子生物学、精确的老鼠疾病模型以及基因的产生和分析方面的专业知识 转录数据。申请者在这个项目中发展的技能将在整个课程中对她有很好的帮助。 她在生物医学研究方面的职业生涯。

PROJECT SUMMARY/ABSTRACT Mutation of the TP53 tumor suppressor gene is the most common genetic alteration in cancer, and over 100 distinct and recurrent missense mutations in TP53 have been identified. Virtually all p53 mutants studied to date have lost the ability to bind to DNA, thereby impairing its function as a transcription factor, and it seems likely that this molecular function largely explains its role in tumor formation. Additionally, many studies have uncovered gain-of-function or oncogenic properties of individual mutants that extend beyond loss of wild type function, most notably the ability to promote invasion and metastasis. Nevertheless, which specific TP53 mutations drive differential tumor development and the molecular mechanisms responsible for these phenotypes remain poorly understood. Given that p53 is a transcription factor and that a significant fraction of cancer- associated mutations, including C132Y, occur in the DNA binding domain, I hypothesize that p53 mutants promote differential tumor formation and progression by profoundly altering the cellular transcriptome in a mutant-specific manner. Aim 1: Characterize the metastatic potential of p53 C132Y mutant. In preliminary studies, I identified the cancer associated mutation, C132Y, as an allele that has greater metastatic potential than other mutants. In this aim, I propose in vitro and in vivo functional characterization of this mutant with the goal of determining: 1) whether the mutant is necessary for metastasis and 2) the molecular mechanisms by which the mutant promotes metastasis. Aim 2: Systematic characterization of p53 missense mutants. Many studies, including some from our own lab, have produced data to suggest that there may be differences in the function of unique missense mutant p53 proteins. What these functions are and to what extent they are conserved between mutants is still poorly understood. To address this question, I propose: 1) a systematic characterization of the transcriptome of cells harboring different mutant p53 proteins and 2) a detailed characterization of the top two mutants from each transcriptional cluster to probe the mechanisms by which the mutants are functioning. Training Plan: The applicant will work with an interdisciplinary team of mentors and collaborators to gain expertise in cell and molecular biology, precision mouse models of disease, and generation and analysis of transcriptomic data. The skills that the applicant will develop during this project will serve her well over the course of her career in biomedical research.