Rational targeting of protein translation of cancer treatments

癌症治疗的蛋白质翻译的合理靶向

基本信息

批准号：
8560772
负责人：
France Carrier
金额：
$ 31.85万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-01 至 2018-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8560772
关键词：
3&apos Untranslated Regions 5&apos Untranslated Regions Affect Antineoplastic Agents Apoptosis Binding Biochemical Biological Assay Cell Proliferation Cell physiology Cells Cellular Stress Characteristics Colon Carcinoma Computer Assisted DNA Repair Data Docking Dose Down-Regulation Drug Design Drug Targeting Eukaryotic Initiation Factor-4G Feedback Fibroblasts Goals Growth Hematologic Neoplasms Heterogeneous-Nuclear Ribonucleoproteins Histocytochemistry Human Lead Malignant - descriptor Malignant Neoplasms Malignant neoplasm of prostate Mantle Cell Lymphoma Metastatic Melanoma Molecular Molecular and Cellular Biology Mus NMR Spectroscopy Normal Cell Normal tissue morphology Nuclear Magnetic Resonance Nucleic Acids Pathway interactions Pharmaceutical Preparations Phosphorylation Proliferating Protein Translation Pathway Proteins RNA RNA Binding RNA-Binding Proteins RNA-Protein Interaction Radiation Regulatory Pathway Renal Cell Carcinoma Research Ribonucleoproteins Role Solid Solid Neoplasm Solutions Specificity Stream Stress Structure Technology Testing Toxic effect Transcript Translations Tuberous sclerosis protein complex Untranslated Regions Western Blotting Work Xenograft Model Xenograft procedure Yang anticancer treatment base cancer cell cancer therapy clinically relevant design drug discovery human tissue inhibitor/antagonist innovation mTOR protein malignant breast neoplasm melanocyte melanoma novel oncology prevent public health relevance response small molecule stem structural biology three dimensional structure tumor tumor growth tumor progression

项目摘要

DESCRIPTION (provided by applicant): Targeting protein translation for cancer therapy is an attractive strategy to limit cancer cells proliferation by depriving them of essential nutriments. However, the currently available drugs target components of the protein translational machinery that are also essential for normal cell functions. Here, we propose to develop a more rational approach by targeting a regulator of protein translation in cancer cells. Our preliminary data indicate that the stress-activated RNA binding protein hnRNP A18 confers growth advantage to cancer cells by regulating the expression of key proteins that prevent apoptosis and/or increase DNA repair. hnRNP A18 increases protein translation by binding to a specific RNA signature motif in the 3'UTRs of its targeted transcripts and interacting with the general translational machinery. Therefore, targeting hnRNP A18 could prevent protein translation in cancer cells, where hnRNP A18 is over expressed, and would only affect the translation of transcripts harboring hnRNP A18 signature motif. Our working hypothesis is that rational targeting of hnRNP A18 will inhibit the translation of specific RNA transcripts devoted to confer growth advantages to cancer cells. To test this hypothesis three specific Aims have been designed. Aim 1: Determine the role of hnRNP A18 in cancer cells sensitivity to anticancer treatments. Levels of hnRNP A18 will be manipulated and the cellular sensitivity to clinically relevant doses of anticancer drugs or radiation will be analyzed by clonogenic survival assays, apoptosis and DNA repair (?H2AX). Aim 2: Characterize the biochemical and functional binding activity of hnRNP A18 to its targeted transcripts and evaluate the effect of hnRNP A18 on cancer progression. This will be performed by systematically analyzing binding to the different stems and bulges of the hnRNP A18 RNA motif and evaluating the effect of hnRNP A18 phosphorylation on the binding to these different RNA structures. Aim 3: Determine the three-dimensional structure of hnRNP A18 and identify the hnRNP A18-RNA binding interface. The structures of unphosphorylated and phosphorylated hnRNP A18 in the absence (apo) and presence of 3'UTR RNA (signature motif) will be solved in solution by Nuclear Magnetic Resonance (NMR) spectroscopy. Our long term goal is to develop drugs that could target hnRNP A18 to control and possibly stop cancer progression. This could be accomplished by solving hnRNP A18 three dimensional structure and use computer-aided drug design (CADD) technology to identify small molecules that could target hnRNP A18.

描述（由申请人提供）：用于癌症治疗的靶向蛋白质翻译是一种通过剥夺癌细胞必需营养来限制癌细胞增殖的有吸引力的策略。然而，目前可用的药物针对蛋白质翻译机制的成分，这些成分对于正常细胞功能也至关重要。在这里，我们建议通过靶向癌细胞中蛋白质翻译的调节因子来开发一种更合理的方法。我们的初步数据表明，应激激活的 RNA 结合蛋白 hnRNP A18 通过调节防止细胞凋亡和/或增加 DNA 修复的关键蛋白的表达，赋予癌细胞生长优势。 hnRNP A18 通过与其目标转录物 3'UTR 中的特定 RNA 特征基序结合并与一般翻译机制相互作用来增加蛋白质翻译。因此，靶向 hnRNP A18 可以阻止癌细胞中的蛋白质翻译，而癌细胞中 hnRNP A18 过度表达，并且只会影响含有 hnRNP A18 特征基序的转录本的翻译。我们的工作假设是，合理靶向 hnRNP A18 将抑制专门赋予癌细胞生长优势的特定 RNA 转录本的翻译。为了检验这一假设，设计了三个具体目标。目标 1：确定 hnRNP A18 在癌细胞抗癌治疗敏感性中的作用。将操纵 hnRNP A18 的水平，并通过克隆存活测定、细胞凋亡和 DNA 修复 (?H2AX) 来分析细胞对临床相关剂量的抗癌药物或放射的敏感性。目标 2：表征 hnRNP A18 与其靶向转录本的生化和功能结合活性，并评估 hnRNP A18 对癌症进展的影响。这将通过系统分析与 hnRNP A18 RNA 基序的不同茎和凸起的结合并评估 hnRNP A18 磷酸化对与这些不同 RNA 结构的结合的影响来进行。目标3：确定hnRNP A18的三维结构并识别hnRNP A18-RNA结合界面。在不存在 (apo) 和存在 3'UTR RNA（特征基序）的情况下，未磷酸化和磷酸化 hnRNP A18 的结构将通过核磁共振 (NMR) 光谱在溶液中解析。我们的长期目标是开发能够靶向 hnRNP A18 的药物，以控制并可能阻止癌症进展。这可以通过解析 hnRNP A18 的三维结构并使用计算机辅助药物设计 (CADD) 技术来识别可以靶向 hnRNP A18 的小分子来实现。