RNA-Binding Proteins as Molecular Integrators that Control the Response of HGSOC to Ant-Cancer Therapies

项目摘要 卵巢癌是美国女性第五大致死性癌症。尽管取得了进步， 在过去几十年的手术切除和铂-紫杉烷联合治疗中，治愈率 保持在相对较低的水平(~30%)，大多数被诊断为晚期卵巢癌的女性将死于 5年内出现耐药疾病。该项目的长期目标是将RNA结合蛋白作为分子 控制HGSOC对抗癌治疗反应的整合子“，是为了识别特定的RNA结合 与其上游的蛋白激酶调节器一起，控制着对 高级别浆液性卵巢癌到这些临床使用的一线抗癌治疗。这项计划涉及： (1)详细的计算分析，使用以下方法查询预先存在的公开可用的RNA表达数据 RNA-BP识别基序用于识别其mRNA靶标为上调或下调的特定RNA结合蛋白 卵巢癌化疗敏感与耐药肿瘤患者的调控；以及(2)一种 RNA-Bps的独立CRISPR干扰和CRISPR激活全基因组筛查 在体外，操纵改变卵巢癌细胞对铂和紫杉烷类药物的耐药性和敏感性， 并在体内用异种细胞系和人PDX建立卵巢癌小鼠模型。RNA-BPS识别器 通过这两种互补的方法，再加上我们已经确定的一组RNA-BP 在以前的实验和初步数据中，直接验证了它们对耐药性的影响 这些模型系统及其结合的RNA的同一性和它们对基因表达的影响被确定 使用片段技术和RNA-Seq.在选定的案例中，特定的磷酸化位点的重要性 通过检测RNA-BP的功能，进一步阐明抗癌药物耐药的分子基础 RNA-BP作用的路径特异性调控。该项目建立在广泛的专业知识和 所有协查实验室的相关工作。这些研究的预期结果包括 识别特定的RNA-Bps和上游调节激酶通路，其靶向可以防止或 逆转卵巢癌对当前临床使用的前列环素疗法的耐药性；阐明 控制化疗后癌症基因表达的新分子电路；以及创造 整个科学界可以使用的一套基于Web的工具，可以用来查询任何一组 基于RNA-BP的调控的差异表达基因，特别是以优化分析的形式 新的和现有的癌症患者RNA表达数据集。

Project Summary Ovarian carcinoma is the fifth deadliest cancer among women in the United States. In spite of advances in surgical resection and platinum-taxane combination therapy over the past several decades, cure rates remain relatively low (~30%) and a majority of women diagnosed with advanced ovarian cancer will die with drug-resistant disease within 5 years. The long-term goal of this project, “RNA-Binding Proteins as Molecular Integrators that Control the Response of HGSOC to Anti-Cancer Therapies”, is to identify specific RNA-binding proteins that, together with their upstream protein kinase regulators, control the resistance and sensitivity of high-grade serous ovarian cancers to these clinically used first line anti-cancer therapies. The project involves: (1) a detailed computational analysis that queries pre-existing publically available RNA expression data using RNA-BP recognition motifs to identify specific RNA binding proteins whose mRNA targets are up- or down- regulated in ovarian cancer patients with chemo-sensitive versus chemo-resistant tumors; and (2) an independent CRISPR-interference and CRISPR-activation genome-wide screen for RNA-BPs whose manipulation alters the resistance and sensitivity of ovarian cancer cells to platinum and taxane agents in vitro, and in vivo using cell line xenografts and human PDX ovarian cancer mouse models. The RNA-BPs identify by these two complimentary approaches, together with a collection of RNA-BPs that we have already identified in previous experiments and preliminary data, are then directly validated for their effects on drug resistance in these model systems, and the identity of their bound RNAs and their effects on gene expression determined using CLIP technologies and RNA-Seq. In selected cases the importance of specific phosphorylation sites on RNA-BP function is examined to further elucidate the molecular basis for anti-cancer drug resistance through pathway-specific regulation of RNA-BP action. The project builds on a broad foundation of expertise and related work from all of the co-Investigators laboratories. Expected outcomes from the studies include the identification of specific RNA-BPs and upstream regulatory kinase pathways whose targeting can prevent or reverse the resistance of ovarian cancers to current clinically used front-lime therapeutics; the elucidation of new molecular circuits that control gene expression in cancers after chemotherapy treatment; and the creation of a suite of web-based tools available to the entire scientific community that can be used to query any set of differentially expressed genes for RNA-BP-based regulation, particularly in a form that is optimized for analysis of new and existing cancer patient RNA expression datasets.