Regulation and Function of the p14ARF/topoisomerase I Complex in Cancer

p14ARF/拓扑异构酶 I 复合物在癌症中的调节和功能

基本信息

批准号：
7813636
负责人：
RUTH A GJERSET
金额：
$ 31.32万
依托单位：
TORREY PINES INST FOR MOLECULAR STUDIES
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2012-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7813636
关键词：
Affect Antibodies Biochemical Biological Biological Assay C-terminal CDKN2A gene Camptothecin Cancer cell line Cell Culture Techniques Cell Line Cells Chromatin Clinic Clinical Coenzyme A Complex DNA DNA Binding DNA Double Strand Break Defect Diagnostic Double Strand Break Repair Down-Regulation Drug Delivery Systems Event Freezing Frequencies Funding Goals Grant Histone Acetylation Human Immunofluorescence Immunologic Immunoprecipitation In Vitro Lead Malignant Neoplasms Maps Mediating Modeling Molecular Molecular Biology Techniques Mutation Nonhomologous DNA End Joining Nude Mice Outcome Pathway interactions Pensions Pharmaceutical Preparations Phosphorylation Phosphorylation Site Play Protein Kinase C Proteins Recovery Regulation Resistance Role Serine Small Interfering RNA Specificity Specimen Spermidine Surrogate Markers TP53 gene Therapeutic Treatment Protocols Tumor Suppressor Proteins Type I DNA Topoisomerases United States National Institutes of Health base cancer cell cancer therapy casein kinase II chemotherapeutic agent chemotherapy histone acetyltransferase human TOP1 protein improved in vitro Assay in vivo inhibitor/antagonist insight neoplastic cell novel novel strategies novel therapeutics overexpression parent grant preclinical evaluation public health relevance repaired research study response subcutaneous tool tumor

项目摘要

DESCRIPTION (provided by applicant): This application responds to Notice Number (NOT-OD-09-058) entitled: NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications. The long term goal of this study is to develop improved cancer treatments through modulation of topoisomerase I (topo I), an important target for camptothecin-like chemotherapeutic drugs, and to further elucidate how the p14ARF/topo I complex contributes to the mechanism of cancer and to the therapy response. p14ARF plays a well-established role in the p53 pathway, and also engages in a novel p53-independent interaction with topo I. The interaction requires topo I serine phosphorylation, activates topo I, and enhances cellular sensitivity to camptothecin and related topo I-targeted chemotherapeutic drugs. Cancer cells with reduced topo I phosphorylation lack p14ARF/topo I complexes and are resistant to camptothecin. A better understanding of the regulation of this complex would therefore elucidate the roles of both proteins in cancer, and could lead to improved diagnostic and therapeutic strategies for cancer. The Specific Aims of the parent grant are to (1) Determine the molecular features of the p14ARF/topo I interaction, (2) Define the molecular mechanism of p14ARF-mediated activation of topo I, (3) Determine how frequently abnormalities in p14ARF/topo I complex formation occur in cancer and whether they correlate statistically with clinic attributes of tumors, and (4) Determine the therapeutic potential of p14ARF-mediated therapy sensitization. The revised grant introduces an additional Aim (5) that studies the effects of Spermidine-CoA-based inhibitors of histone acetylation, a class of cancer cell-targeted compounds that inhibit double strand break repair and sensitize cells to camptothecin. The Aim examines how these inhibitors affect topo I alone and in combination with p14ARF and provides a preclinical evaluation of their efficacy as therapy sensitizers in the presence and absence of ARF. The study will employs molecular biology techniques (Aim I), biochemical assays (Aims II and V), immunoprecipitation/western analysis and immunofluorescence (Aim III), and cell culture-based viability assays and human tumor xenograph models in nude mice (Aims IV and V). PUBLIC HEALTH RELEVANCE: This study develops novel approaches based on the p14ARF tumor suppressor and Spermidine-CoA-based inhibitors of histone acetylation to improve cellular responses to topoisomerase I-targeted chemotherapies and reverse therapy resistance, a major obstacle to successful cancer treatment. The study also develops diagnostic tools for identifying therapy responsive tumors and optimizing therapeutic regimens. .

描述（由申请人提供）：本申请响应通知号（NOT-OD-09-058），标题为：NIH 宣布恢复法案基金可用于竞争性修订申请。这项研究的长期目标是通过调节拓扑异构酶 I (topo I)（喜树碱类化疗药物的重要靶点）开发改进的癌症治疗方法，并进一步阐明 p14ARF/topo I 复合物如何促进癌症机制以及治疗反应。 p14ARF 在 p53 通路中发挥着明确的作用，并且还参与与拓扑 I 的新型独立于 p53 的相互作用。该相互作用需要拓扑 I 丝氨酸磷酸化，激活拓扑 I，并增强细胞对喜树碱和相关拓扑 I 靶向的敏感性化疗药物。 topo I 磷酸化降低的癌细胞缺乏 p14ARF/topo I 复合物，并且对喜树碱具有抗性。因此，更好地了解这种复合物的调节将阐明这两种蛋白质在癌症中的作用，并可能导致改进癌症的诊断和治疗策略。母基金的具体目标是 (1) 确定 p14ARF/topo I 相互作用的分子特征，(2) 定义 p14ARF 介导的 topo I 激活的分子机制，(3) 确定 p14ARF/ 中异常的频率topo I 复合物的形成发生在癌症中，以及它们是否与肿瘤的临床属性有统计相关性，以及 (4) 确定 p14ARF 介导的治疗敏化的治疗潜力。修订后的拨款引入了另一个目标 (5)，研究基于亚精辅酶 A 的组蛋白乙酰化抑制剂的作用，这是一类靶向癌细胞的化合物，可抑制双链断裂修复并使细胞对喜树碱敏感。该目的研究了这些抑制剂如何单独影响拓扑 I 以及与 p14ARF 联合使用，并提供其在存在和不存在 ARF 的情况下作为治疗增敏剂的功效的临床前评估。该研究将采用分子生物学技术（Aim I）、生化测定（Aims II 和 V）、免疫沉淀/蛋白质印迹分析和免疫荧光（Aim III）以及基于细胞培养的活力测定和裸鼠人体肿瘤异种移植模型（Aims IV）和五）。公共健康相关性：本研究开发了基于 p14ARF 肿瘤抑制剂和基于亚精辅酶 A 的组蛋白乙酰化抑制剂的新方法，以改善细胞对拓扑异构酶 I 靶向化疗的反应并逆转治疗耐药性，这是成功癌症治疗的主要障碍。该研究还开发了用于识别治疗反应性肿瘤和优化治疗方案的诊断工具。。