Therapeutic implication of estrogen receptor-p53 interaction in mitochondria

线粒体中雌激素受体-p53 相互作用的治疗意义

基本信息

批准号：
8435384
负责人：
GOKUL M. DAS
金额：
$ 18.99万
依托单位：
ROSWELL PARK CANCER INSTITUTE CORP
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-03-01 至 2016-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8435384
关键词：
Address Affect Apoptosis Apoptotic Bees Binding Biological Assay Biology Breast Cancer Cell Cell Nucleus Complement Complex Defect Detection Diagnosis Disease Progression Epithelial Estrogen Receptor 1 Estrogen Receptors Estrogens Frequencies Funding Gene Expression Gene Targeting Genes Genetic Transcription Goals Human Immunoprecipitation Intervention Laboratories Lead Malignant Neoplasms Mammary Neoplasms Mammary gland Metabolism Mission Mitochondria Molecular Analysis Mutate Mutation Nuclear Nuclear Proteins Oncogenes Oncogenic Outcome Oxidative Phosphorylation Pathway interactions Play Prevention Preventive Intervention Protein p53 Proteins Public Health RNA Interference Reporting Research Respiration Role Seminal Signal Transduction Stem cells TP53 gene Testing Therapeutic Therapeutic Intervention Time Transfection Tumor Suppressor Proteins Work cancer epidemiology cell growth regulation complex IV cytochrome c oxidase human disease innovation insight malignant breast neoplasm new therapeutic target novel novel therapeutics outcome forecast promoter research study respiratory response stem tumor xenograft

项目摘要

DESCRIPTION (provided by applicant): Estrogen receptor-1 (ER) and tumor suppressor p53 play important, but opposite, roles in the onset and progression of breast cancer. Compared to other cancers, overall frequency of p53 mutation in breast cancer is about 20%; however, wild type p53 is functionally debilitated. Both ER and p53 are localized in the nuclei as well as mitochondria and are functionally important in both the compartments. ER has been reported to bind and inhibit wild type p53 function in the nucleus. There is a fundamental gap in understanding how mitochondrial p53 is antagonized by ER. This gap represents an important problem because deciphering the role of ER in suppressing mitochondrial p53 is essential in understanding the mechanisms by which wild type p53 is inactivated in breast cancer. The long-term goal is to understand the mechanisms by which nuclear and mitochondrial functions of p53 are compromised in breast cancer. The objective in this particular application is to analyze mitochondrial ER-p53 interaction and its functional consequences. The central hypothesis is that ER and p53 interact within mitochondria leading to important functional consequences. The rationale that underlies the proposed research is that given the importance of mitochondrial p53 in responding to oncogenic signaling and regulation of cellular metabolism, understanding the interaction between mitochondrial ER and p53 and its functional consequences would provide new therapeutic targets in addition to mechanistic insights that could be exploited for better intervention strategies. This hypothesis wil be tested by pursuing three specific aims: 1) Analyze ER-p53 interaction in mitochondria in breast cancer cells; 2) Investigate effect of ER- p53 interaction on oxidative phosphorylation (OXPHOS) in mitochondria; and 3) Determine the effect of ER- p53 interaction on mitochondrial gene transcription and apoptosis. Under the first aim, already proven immunoprecipitation (IP) assay, RNA interference (RNAi) approach, and gene transfection approaches, which have been established in the applicants' laboratories, will be used to characterize ER-p53 interaction within mitochondria. Under the second specific aim, effect of ER-p53 interaction on OXPHOS will be analyzed using approaches such as respiratory complex assays and quantitative real-time PCR (qPCR) assay that are established in the applicants' laboratories along with a OXPHOS-Chip expression array already developed and validated by the applicants. The third aim will address, using multiple technical approaches already optimized by the applicants, how ER-p53 interaction affect mitochondrial gene transcription and apoptosis. The research is innovative because it represents a new and substantive departure from the status quo, namely analyzing ER and p53 function in breast cancer cell mitochondria in an integrated manner instead of pursuing them as independent regulators of separate pathways. The proposal is significant because it is the first step in a continuum of research that is expected to lead to a better understanding of the roles of mitochondrial ER and p53 in breast cancer that could be exploited for developing new therapeutic strategies.

描述（由申请人提供）：雌激素受体-1（ER）和肿瘤抑制p53在乳腺癌的发作和进展中起重要但相反的作用。与其他癌症相比，乳腺癌中p53突变的总频率约为20％。但是，野生型p53在功能上会衰弱。 ER和p53均位于细胞核以及线粒体中，并且在两个隔室中都在功能上很重要。据报道，ER在细胞核中结合并抑制野生型p53功能。理解线粒体p53是如何通过ER拮抗的基本差距。该差距代表了一个重要的问题，因为解密ER在抑制线粒体p53中的作用对于理解野生型p53在乳腺癌中灭活的机制至关重要。长期目标是了解p53核和线粒体功能在乳腺癌中受到损害的机制。该特定应用的目的是分析线粒体ER-P53相互作用及其功能后果。中心假设是ER和p53在线粒体内相互作用，从而导致重要的功能后果。拟议的研究基础的基本原理是，鉴于线粒体p53在响应致癌信号和细胞代谢的调节中的重要性，因此了解线粒体ER和p53及其功能后果之间的相互作用及其功能后果还可以提供新的治疗目标，而除了可以更好地介绍机械化培训以获得更好的互动介入，可以利用机械性介入。该假设将通过追求三个特定目的来检验：1）分析乳腺癌细胞中线粒体中的ER-P53相互作用； 2）研究ER-P53相互作用对线粒体中氧化磷酸化（OXPHOS）的影响； 3）确定ER-P53相互作用对线粒体基因转录和凋亡的影响。在第一个目标下，已经在申请人实验室中确定的已经被证明的免疫沉淀（IP）测定，RNA干扰方法（RNAI）方法和基因转染方法将用于表征线粒体内的ER-P53相互作用。在第二个特定目的下，将使用诸如申请人实验室中确定的呼吸复合物测定和定量实时PCR（QPCR）测定法以及OXPHOS-CHIP表达阵列在已经开发和验证的OXPHOS-CHIP表达阵列以及申请人验证的方法中分析ER-P53相互作用对OXPHOS的影响。第三个目标将使用申请人已经优化的多种技术方法来解决ER-P53相互作用如何影响线粒体基因转录和凋亡。这项研究具有创新性，因为它代表了与现状的新事物，即以综合方式分析乳腺癌细胞线粒体中的ER和p53功能，而不是追求它们作为单独途径的独立调节剂。该提议很重要，因为这是一项连续研究的第一步，预计将更好地理解线粒体ER和p53在乳腺癌中的作用，这些角色可以用于开发新的治疗策略。