Attacking aggressive p53 mutants in gynecologic cancer

项目总结/摘要 我目前是一个指导助理研究科学家在妇产科在 爱荷华州大学。我的长期目标是建立一个独立的研究项目，以改善结果 为患有卵巢癌和子宫内膜癌的女性提供安全有效的个性化治疗 治疗方案。我打算在一个竞争激烈的研究机构寻求一个独立的终身教职。 机构，建立在我的博士后培训在新的RNA为基础的治疗和目前的工作， 针对妇科癌症的个性化药物。我的独立K22研究计划的目标是 机械地区分常见的错义p53突变体，并开发基于合成RNA的 治疗剂来克服有害功能。在所有观察到的p53突变中，约有40-50%是 单核苷酸变异导致改变单个氨基酸的错义突变。错义 突变不仅消除了典型的DNA结合和与辅因子的相互作用， 活性，包括非典型靶点的转录和蛋白质：蛋白质相互作用中的新活性。研究将 由于p53突变在子宫内膜癌和卵巢癌中的广泛发生， p53突变型癌症患者的5年生存率极低。我假设这种误解 导致蛋白质超稳定的p53突变激活不同的转录组特征， 使用新的RNA适体干扰，进化为对每个突变体p53特异。这项工作利用了一个高度 从患者源性类器官（PDO）培养物中分离天然p53的创新策略， p53基因突变的可能性。首先，我将确定生存机制， 通过计算DNA结合特异性天然p53突变体的复发性p53突变体的化学抗性， 将其与基因组定位、基因调控和化学敏感性联系起来。这项工作包括一种新的方法， 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p53突变体模型的化疗敏感性： 用突变体特异性RNA适体干扰单个p53突变体的有害活性。这些研究 将提供第一个全面评估的功能后果的一个大面板的p53 突变体，并建立开发基于适体的工具， 变种人这个项目融合了我在细胞信号传导方面的研究生工作， 治疗学和最近在妇科癌症转化研究中的工作。在K22的资助下 获奖和我的多学科合作者，我将有能力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.