Understanding the divergent functions of mutant p53

了解突变体 p53 的不同功能

基本信息

批准号：
10661541
负责人：
Margaret Kennedy
金额：
$ 4.77万
依托单位：
SLOAN-KETTERING INST CAN RESEARCH
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：
2022-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10661541
关键词：
Address Adopted Affect Alleles Binding Biological Assay Biomedical Research Cancer Patient Cell Line Cell physiology Cells Cellular biology ChIP-seq Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Coupled Couples DNA DNA Binding Domain Data Development Disease model Doxycycline Ductal Epithelial Cell Engineering Gene Chips Gene Expression Generations Genetic Transcription Goals Guide RNA Human Impairment In Vitro Individual Invaded Laboratories Malignant Neoplasms Mentors Missense Mutation Molecular Molecular Biology Mus Mutate Mutation Neoplasm Metastasis Oncogenic Pancreatic Ductal Adenocarcinoma Pancreatic duct Pathway interactions Patients Pattern Phenotype Property Proteins Publishing Recurrence Renilla Reporting Role Sampling Sorting TP53 gene Testing The Cancer Genome Atlas Tissue-Specific Gene Expression Training Tumor Promotion Tumor Suppressor Genes Tumor-Derived Variant Work base editing biobank career chromatin immunoprecipitation gain of function in vivo metastatic process mouse model mutant novel strategies pancreatic cancer patients pancreatic ductal adenocarcinoma model pancreatic neoplasm premalignant programs single-cell RNA sequencing skills small hairpin RNA transcription factor transcriptome transcriptome sequencing transcriptomic profiling transcriptomics treatment strategy tumor tumorigenic virtual

项目摘要

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.

项目概要/摘要 TP53 肿瘤抑制基因的突变是癌症中最常见的基因改变，超过 100 TP53 中明显且反复出现的错义突变已被鉴定。迄今为止研究的几乎所有 p53 突变体失去了与 DNA 结合的能力，从而损害了其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，许多研究还发现单个突变体的功能获得或致癌特性，其范围超出了野生型的丧失功能，最显着的是促进侵袭和转移的能力。尽管如此，具体的TP53 突变驱动不同的肿瘤发展以及导致这些表型的分子机制仍然知之甚少。鉴于 p53 是一种转录因子，并且很大一部分癌症- 相关突变，包括 C132Y，发生在 DNA 结合域中，我假设 p53 突变体通过深刻改变细胞转录组来促进差异性肿瘤的形成和进展突变体特异性方式。目标 1：表征 p53 C132Y 突变体的转移潜力。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体具有更大转移潜力的等位基因。在这个目的，我建议对该突变体进行体外和体内功能表征，目的是确定：1) 突变体是否是转移所必需的；2）突变体促进转移的分子机制转移。目标 2：p53 错义突变体的系统表征。许多研究，包括我们自己的一些研究实验室产生的数据表明，独特的错义突变体 p53 的功能可能存在差异蛋白质。这些功能是什么以及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）细胞转录组的系统表征含有不同的突变体 p53 蛋白，2) 每个突变体中前两个突变体的详细特征转录簇来探测突变体发挥作用的机制。培训计划：申请人将与由导师和合作者组成的跨学科团队合作，以获得细胞和分子生物学、精准小鼠疾病模型以及生成和分析的专业知识转录组数据。申请人在该项目中培养的技能将对她在整个课程中受益匪浅她的生物医学研究生涯。