Understanding the divergent functions of mutant p53

了解突变体 p53 的不同功能

• 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
• 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

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.

项目摘要/摘要 TP53肿瘤抑制基因的突变是癌症中最常见的遗传改变，超过100 已经确定了TP53中明显的反复错义突变。迄今为止研究的所有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，并且似乎很可能 该分子功能在很大程度上解释了其在肿瘤形成中的作用。此外，许多研究都有 单个突变体的未发现功能获得或致癌特性，这些突变体超出了野生型的损失 功能，最著名的是促进入侵和转移的能力。但是，哪个特定的TP53 突变驱动差异肿瘤的发展和负责这些表型的分子机制 保持不当理解。鉴于p53是转录因子，很大一部分癌症 相关的突变（包括C132Y）发生在DNA结合结构域中，我假设p53突变体 通过深刻改变A中的细胞转录组来促进差异肿瘤的形成和进展 突变特异性的方式。 AIM 1：表征p53 C132Y突变体的转移潜力。在初步研究中，我确定了 与其他突变体相比，癌症相关的突变C132Y是具有更大转移潜力的等位基因。在这个 目的，我提出了该突变体的体外和体内功能表征，其目的是确定：1​​） 突变体是否需要转移和2）突变体促进的分子机制 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 实验室产生了数据，以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，在突变体之间它们在多大程度上仍然很差 理解。为了解决这个问题，我提出：1）细胞转录组的系统表征 携带不同的突变体p53蛋白和2）每个来自每个突变体的详细表征 转录簇以探测突变体发挥作用的机制。 培训计划：申请人将与跨学科的导师和合作者合作，以获得 细胞和分子生物学的专业知识，疾病的精确小鼠模型以及生成和分析 转录组数据。申请人在此项目期间将发展的技能将在整个课程中为她服务 她从事生物医学研究的职业。