Attacking aggressive p53 mutants in gynecologic cancer

项目摘要/摘要 我目前是美国国立卫生研究院妇产科的助教研究科学家。 爱荷华大学。我的长期目标是建立一个独立的研究计划，以改善结果 为卵巢癌和子宫内膜癌患者提供安全、有效和个性化的 治疗方案。我打算在一家竞争性研究机构寻求一份独立的终身教职。 机构，基于我在基于RNA的新疗法方面的博士后培训和目前在 妇科癌症的个体化用药。我的独立K22研究项目的目标是 机械区分常见的p53错义突变体并开发基于合成RNA的 治疗，克服有害的功能。在所有观察到的p53突变中，约有40%-50%是 导致错义突变从而改变单一氨基酸的单核苷酸变体。误会 突变不仅消除了规范的DNA结合和与辅助因子的相互作用，而且还赋予了新的 活动，包括非规范靶标的转录和新的蛋白质：蛋白质相互作用。研究将会 鉴于p53基因突变在子宫内膜癌和卵巢癌中的广泛存在，应重点关注这些癌症 P53突变癌症患者的类型和极差的5年生存率。我假设这是个误会 导致蛋白质超稳定的p53突变激活了不同的转录信号，这些信号可能是 使用新型RNA的干扰适配子进化为对每个突变型p53具有特异性。这项工作充分利用了 从患者衍生的器官(PDO)培养中分离原生p53的创新策略，从而允许研究 在他们的内源环境中发生的p53突变。首先，我要确定生存和发展的机制(S) 通过计算DNA结合特异性、天然p53突变体和 这与基因组定位、基因调控和化疗敏感性有关。这项工作包括一种新的方法来 从PDO模型中分离p53突变蛋白以捕获天然蛋白构象，翻译后 修饰和剪接异构体。接下来，我将通过以下方法恢复带有p53突变的模型的化疗敏感性 用突变型特异性RNA适配子干扰单个突变型p53的有害活性。这些研究 将首次对一大批p53的功能后果进行全面评估 使用天然蛋白的突变体，并建立基于适体的工具来灭活P53的可行性 变种人。这个项目融合了我在细胞信号方面的研究生工作，以及基于RNA适体的博士后培训 治疗学和妇科癌症的翻译研究的最新工作。在K22的资助下 奖和我的多学科合作者，我将有条件建立一个独立的研究 该计划旨在改善妇科癌症患者的预后。

Project Summary/Abstract I am currently a mentored assistant research scientist in the Department of Obstetrics and Gynecology at the University of Iowa. My long-term goal is to establish an independent research program to improve outcomes for women with ovarian and endometrial cancer through the safe delivery of effective and personalized treatment regimens. I intend to pursue an independent tenure-track faculty position at a competitive research institution, building upon my postdoctoral training in novel RNA-based therapeutics and current work in personalized medicine for gynecologic cancers. The objective for my independent K22 research program is to mechanistically distinguish between common missense p53 mutants and develop synthetic RNA-based therapeutics to overcome the deleterious functions. Approximately 40-50% of all observed p53 mutations are single nucleotide variants that result in missense mutations that change a single amino acid. Missense mutations not only abrogate canonical DNA binding and interaction with co-factors, but also confer new activities, including transcription of non-canonical targets and new in protein:protein interactions. Studies will focus on endometrial and ovarian cancer given the widespread occurrence of p53 mutations in these cancer types and the extremely poor 5-year survival of patients with p53 mutant cancers. I hypothesize that missense mutations in p53 that result in protein hyperstabilization activate different transcriptomic signatures that can be perturbed using novel RNA aptamers evolved to be specific for each mutant p53. This work leverages a highly innovative strategy to isolate native p53 from patient-derived organoid (PDO) cultures, which allows for study of p53 mutations in their endogenous environment. First, I will determine the mechanism(s) of survival and chemoresistance for recurrent p53 mutants by calculating the DNA binding specificity native p53 mutants and link this to genomic localization, gene regulation and chemosensitivity. This work includes a novel approach to isolate p53 mutant proteins from PDO models to capture the native protein conformation, posttranslational modifications and splice isoforms. Next, I will restore chemosensitivity in models with p53 mutants by perturbing the deleterious activity of individual p53 mutants with mutant-specific RNA aptamers. These studies will provide the first comprehensive assessment of the functional consequences of a large panel of p53 mutants using native protein and establish the feasibility of developing aptamer-based tools that inactivate p53 mutants. This project merges my graduate work in cell signaling, postdoctoral training in RNA aptamer-based therapeutics and more recent work in translational studies of gynecologic cancers. With funding from this K22 award and my multi-disciplinary collaborators, I will be well-equipped to establish an independent research program to improve outcomes for women with gynecologic cancer.