Mitochondria in Prostate Cancer Diversity

前列腺癌中的线粒体多样性

基本信息

批准号：
9901469
负责人：
KESHAV K SINGH
金额：
$ 33.63万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9901469
关键词：
AR gene Address Affect African American Age American Androgen Receptor Androgens Apoptosis Applications Grants Basic Cancer Research Bioinformatics Cell Line Cell Nucleus Cells Complex DNA DNA Binding Domain Development Diagnosis Disease Electron Transport Complex III Epithelial Cells Family Genes Genetic Genetic Transcription Germ-Line Mutation Green Fluorescent Proteins Inherited Malignant Neoplasms Malignant neoplasm of prostate Missense Mutation Mitochondria Mitochondrial DNA Mitochondrial Proteins Mothers Mus Mutation N Domain N-terminal Neoplasm Metastasis Nuclear Nuclear Localization Signal Oxidative Phosphorylation PC3 cell line Play Ploidies Polymorphism Analysis Prostate Prostatic Neoplasms Proteins Race Recording of previous events Role Signal Transduction Testing Thinking Time Tissues Tumor Volume Tumor stage Variant Xenograft Model base cancer health disparity caucasian American cell transformation complex IV early onset member men mitochondrial genome mortality mutant novel oligomycin sensitivity-conferring protein prognostic significance prostate cancer cell prostate cancer metastasis prostate cancer progression prostate carcinogenesis public health relevance racial difference racial disparity racial diversity receptor receptor expression tumor

项目摘要

DESCRIPTION (provided by applicant): The androgen receptor (AR) plays an important role in normal development of the prostate gland, in prostate carcinogenesis, and in the progression of prostate cancer to advanced metastatic disease. Traditional thinking is that AR localizes exclusively to the nucleus and that nuclear AR regulates genes that are essential to prostate cancer development. This is true. However, we demonstrate a previously unrecognized function of AR in mitochondria. We have discovered that AR 1) directly localizes into the mitochondria and 2) indirectly transcriptionally regulates nuclear genes whose products localize into mitochondria and perform mitochondrial functions. Our studies reveal that i) AR localizes into mitochondria in primary prostate tissues and cell lines, ii) AR is imported into isolated mitochondria, and iii) AR contains a mitochondrial localization signal (MLS) capable of targeting foreign proteins, such as green fluorescent protein, into mitochondria. Indirectly, AR controls expression of a variety of nuclear DNA (nDNA)-encoded mitochondrial oxidative phosphorylation (mtOXPHOS) subunits, including NDUFB8 (Complex I), SDHB (Complex II), UQCRC2 (Complex III), COXII subunit (Complex IV), and ATP5A (Complex V). AR also down-regulates the TFAM, GFM1, and GFM2 genes, which control mitochondrial DNA (mtDNA) content. Consistent with this, the mtDNA content and the expression of mtDNA-encoded COX II protein is significantly reduced in PC3-AR prostate cells expressing AR. Notably, we demonstrate that the mtDNA content in prostate tumors of African-Americans (AA) is >6 times less than in tumors of Caucasian-Americans (CA). mtDNA content was also lower in normal prostates of AA than CA. To identify the underlying mitochondrial basis of prostate cancer diversity, we conducted comprehensive, race-based bioinformatics analyses of variants in more than 6000 AA and 33,000 CA and discovered, in the AR gene of AA, missense variants located in two domains: the N-terminal domain containing the MLS and the DNA-binding domain. Of note, missense mutations in CA were found only in the hinge domain containing the nuclear localization signal (NLS) of AR. Expression of AR variant S598G in PC3 cells reduced more than the wild type the expression of TFAM, which controls mtDNA content. We hypothesize that AR missense variants/mutants present, solely in AA, contribute to the "gain" or "loss" of mitochondrial function and thereby to prostate cancer diversity in AA. AIM 1: Determine the prognostic significance of AR missense variants/mutants and mtDNA content on prostate cancer metastasis and reoccurrence in AA and CA. AIM 2: Evaluate the significance of mitochondrial AR missense variants/mutants as direct regulators of mitochondrial functions that affect composition, organization, stability, and activity of mtOXPHOS super-complexes and apoptosis. AIM 3: Evaluate the significance of nuclear AR missense variants/mutants as indirect regulators of mitochondrial function. AIM 4: Use mouse xenograft model to establish the significance of mitochondrial and nuclear AA- and CA-specific AR missense variants/mutants on prostate tumorigenesis and metastasis.

描述（由申请人提供）：雄激素受体（AR）在前列腺的正常发育、前列腺癌发生以及前列腺癌进展为晚期转移性疾病中发挥着重要作用。传统观点认为，AR仅局限于前列腺。然而，我们发现 AR 1) 直接定位于线粒体中。 2) 间接转录调节核基因，其产物定位于线粒体并执行线粒体功能。我们的研究表明，i) AR 定位于原代前列腺组织和细胞系的线粒体中，ii) AR 被导入分离的线粒体中，以及 iii) AR 含有一个线粒体定位信号 (MLS) 能够将外源蛋白（例如绿色荧光蛋白）定位到线粒体中，AR 间接控制多种核 DNA (nDNA) 编码的线粒体氧化磷酸化的表达。 (mtOXPHOS) 亚基，包括 NDUFB8（复合物 I）、SDHB（复合物 II）、UQCRC2（复合物 III）、COXII 亚基（复合物 IV）和 ATP5A（复合物 V）也会下调 TFAM、GFM1 和 GFM2。与此一致的是，mtDNA 含量和 mtDNA 编码的表达。 COX II 蛋白在表达 AR 的 PC3-AR 前列腺细胞中显着减少。值得注意的是，我们证明非裔美国人 (AA) 前列腺肿瘤中的 mtDNA 含量比白种人美国人 (CA) 肿瘤中的 mtDNA 含量低 6 倍以上。正常前列腺中 AA 的含量也低于 CA，为了确定前列腺癌多样性的线粒体基础，我们对 6000 多个 AA 和 CA 的变异进行了全面的、基于种族的生物信息学分析。 33,000 个 CA 并发现，在 AA 的 AR 基因中，错义变体位于两个结构域：包含 MLS 的 N 端结构域和 DNA 结合结构域。值得注意的是，CA 中的错义突变仅在包含 MLS 的铰链结构域中发现。 AR 的核定位信号 (NLS)。PC3 细胞中 AR 变体 S598G 的表达比野生型降低了更多的 TFAM 的表达，而 TFAM 控制 mtDNA 含量。仅在 AA 中，有助于线粒体功能的“获得”或“丧失”，从而影响 AA 中前列腺癌的多样性目的 1：确定 AR 错义变异/突变体和 mtDNA 含量对 AA 中前列腺癌转移和复发的预后意义。和 CA. 目标 2：评估线粒体 AR 错义变体/突变体作为影响 mtOXPHOS 组成、组织、稳定性和活性的线粒体功能直接调节因子的重要性。 AIM 3：评估核 AR 错义变体/突变体作为线粒体功能间接调节因子的重要性。 AIM 4：使用小鼠异种移植模型建立线粒体和核 AA 和 CA 特异性 AR 错义变体的重要性。 /突变体对前列腺肿瘤发生和转移的影响。