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

RNA 结合蛋白作为分子整合剂控制 HGSOC 对抗癌疗法的反应

基本信息

批准号：
10054974
负责人：
CHRISTOPHER B BURGE
金额：
$ 53.65万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-12-07 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10054974
关键词：
Address Antimitotic Agents Antineoplastic Agents Antitumor Response Ants Area Biological Models CRISPR interference Cancer Patient Cell Line Chemoresistance Cisplatin Clinical Clustered Regularly Interspaced Short Palindromic Repeats Collection Combined Modality Therapy Communities Computer Analysis Cytotoxic agent DNA Damage DNA Sequence DNA-dependent protein kinase Data Data Set Defect Development Diagnosis Disease Disease Resistance Drug resistance Epithelial Excision Foundations Gene Expression Gene Expression Profile Genes Genetic Transcription Goals Hematopoietic Neoplasms Histologic Human Immunotherapy In Vitro Laboratories Limes Link Malignant Neoplasms Malignant neoplasm of ovary Measures Mediating Medical Messenger RNA Mitotic Molecular Mutation Non-Small-Cell Lung Carcinoma Operative Surgical Procedures Outcome Study Ovarian Carcinoma Paclitaxel Pathway interactions Patients Pharmaceutical Preparations Phosphorylation Site Phosphotransferases Platinum Platinum Compounds Protein Kinase Proteins RNA RNA Binding RNA Processing RNA Stability RNA interference screen RNA-Binding Proteins Recurrence Regulation Relapse Research Personnel Resistance Role Serous Signal Pathway Solid Neoplasm TP53 gene Technology Therapeutic Treatment Protocols United States Woman Work Xenograft Model Xenograft procedure base bioinformatics pipeline brca gene cancer cell cancer drug resistance cancer therapy chemotherapy cytotoxic differential expression experimental study frontier genome wide screen genome-wide homologous recombination in vivo inhibitor/antagonist molecular targeted therapies mouse model mutant ovarian neoplasm patient derived xenograft model prevent recombinational repair resistance mechanism resistance mutation response screening standard of care success taxane therapeutic development therapy resistant transcriptome sequencing tumor upstream kinase web-based tool

项目摘要

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.

项目概要尽管取得了进展，但卵巢癌仍是美国女性第五大致命癌症。过去几十年手术切除和铂类紫杉烷联合治疗的治愈率仍然相对较低（~30%），大多数被诊断患有晚期卵巢癌的女性将死于该项目的长期目标是“RNA结合蛋白作为分子”。控制 HGSOC 对抗癌疗法反应的整合器”，旨在识别特定的 RNA 结合与其上游蛋白激酶调节因子一起控制细胞的耐药性和敏感性的蛋白质该项目涉及： (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 识别基序可识别 mRNA 靶点上调或下调的特定 RNA 结合蛋白在化疗敏感与化疗耐药肿瘤的卵巢癌患者中受到调节；(2) 独立的 CRISPR 干扰和 CRISPR 激活全基因组筛选 RNA-BP，其体外操作改变了卵巢癌细胞对铂类和紫杉烷类药物的抵抗力和敏感性，并使用细胞系异种移植物和人 PDX 卵巢癌小鼠模型进行体内 RNA-BP 鉴定。通过这两种互补的方法，以及我们已经鉴定的 RNA-BP 集合在之前的实验和初步数据中，然后直接验证它们对耐药性的影响这些模型系统，以及它们结合的RNA的身份及其对基因表达的影响被确定使用 CLIP 技术和 RNA-Seq 在某些情况下特定磷酸化位点的重要性。通过检查RNA-BP功能，进一步阐明抗癌耐药性的分子基础 RNA-BP 作用的途径特异性调控。该项目建立在广泛的专业知识和基础之上。所有联合研究人员实验室的相关工作包括：鉴定特定的 RNA-BP 和上游调节激酶途径，其靶向可以预防或逆转卵巢癌对当前临床使用的前沿疗法的耐药性；控制化疗后癌症基因表达的新分子回路；整个科学界可用的一套基于网络的工具，可用于查询任何一组用于基于 RNA-BP 的调控的差异表达基因，特别是针对分析进行优化的形式新的和现有的癌症患者 RNA 表达数据集。