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Interference with Androgen Receptor and Its Ligands

干扰雄激素受体及其配体

基本信息

批准号：
7963169
负责人：
JAMES L MOHLER
金额：
$ 63.3万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7963169
关键词：
17p Active Sites Acute Adrenal Glands American Androgen Metabolism Androgen Receptor Androgen Therapy Androgens Androsterone Back CYP17A1 gene Castration Cell Death Cells Cholesterol Homeostasis Clinical Clinical Trials Complex Cytochromes Disease remission Drug Formulations Enzymes Family Freezing Gene Expression Glycols Growth Immunohistochemistry In Vitro Infection Institution Instruction Intervention Isoenzymes Laboratories Learning Ligands Malignant neoplasm of prostate Mass Spectrum Analysis Measures Medical Messenger RNA Metabolism Metastatic Neoplasm to the Bone Mountain Lion Mutation Operative Surgical Procedures Oxidoreductase Pathway interactions Patients Pharmaceutical Preparations Pharmacologic Substance Pre-Clinical Model Pregnenolone Progesterone Prostate Proteins Radioimmunoassay Recurrence Research Personnel Source Stanolone Steroids Stress Suspension substance Suspensions Testing Testosterone Time Tissue Sample Tissues Transactivation Xenograft Model Xenograft procedure abiraterone cancer cell cell growth deprivation experience in vivo inhibitor/antagonist killings men novel novel strategies preclinical study prevent response small hairpin RNA small molecule

项目摘要

PROJECT SUMMARY (See instructions): Project 1 has contributed to two paradigm changing discoveries. First, the androgen receptor (AR) pathway appears critical to the growth of castration-recurrent prostate cancer (CaP) in spite of castrate levels of circulating testicular androgens. Second, castration-recurrent CaP produces high tissue levels of testosterone (T) and dihydrotestosterone (DHT), the preferred AR ligand. Intracrine-produced testicular androgens present a target for novel therapies. Further advances may result from applying novel treatments to CaP coincidental with androgen deprivation therapy (ADT) when CaP cell death is at a maximum and surviving cells are under greatest stress. The central hypothesis ofthe proposed studies is that CaP can be cured or remission extended by a coordinated attack upon intracrine androgen metabolism and AR coincidental with elimination of circulating testicular androgens (medical or surgical castration). In order to test this hypothesis and allow the formulation of an appropriate clinical trial in advanced CaP, the response to ADT will be assessed using preclinical models in the immediate post-castration period. In general, cell or tissue sampling will be conducted just prior to castration and 12h and 1, 2, 4, 8, 16 and SOd after "castration." The effect of castration upon the androgen axis will be assessed on 4 levels in vitro and 6 levels in vivo. The 4 levels of assessment in vitro and in vivo will include 1) changes in androgen metabolism enzymes at the mRNA level using qRT-PCR and protein level using immunohistochemistry (IHC); 2) androgen levels using LC-MS; 3) androgen-regulated gene expression using IHC for PSA, NkxS.1 and hK2; and 4) cell growth. Measures in vivo will include 5) time to progression and 6) survival. Androgen metabolism after ADT will be studied using androgen-sensitive CWR22 cell suspensions, the androgen-dependent CWR22 xenograft and a fresh surgical tissue xenograft model. Changes in androgen metabolism critical for survival after ADT will be targeted using shRNA or drug in vitro and lentiviral shRNA and/or drug in vivo (Aim 1). Testicular androgens formed by intracrine metabolism of adrenal androgens will be removed using 53-reductase or Sult2A1 delivered using lentiviral infection (Aim 2). Finally, AR will be removed using lentiviral AR shRNA constructs or AR degrading small molecules (Aim 3).

项目总结（见说明）：项目1促成了两个改变范式的发现。首先，雄激素受体（AR）途径似乎对去势复发性前列腺癌（CaP）的生长至关重要，尽管去势水平循环睾丸雄激素其次，去势复发性CaP产生高组织水平的睾酮（T）和双氢睾酮（DHT），优选的AR配体。内分泌睾丸雄激素是新疗法的靶点。应用新的治疗方法可能会带来进一步的进步当CaP细胞死亡达到最大时，与雄激素剥夺治疗（ADT）同时发生，存活的细胞承受着最大的压力。这项研究的中心假设是，通过对分泌内雄激素代谢和AR的协调攻击而治愈或缓解延长与循环睾丸雄激素的消除（药物或手术去势）同时发生。为了测试这一假设，并允许制定一个适当的临床试验，在先进的CaP，反应将在去势后即刻使用临床前模型评估ADT。一般来说，细胞或将在去势前和去势后12小时、1、2、4、8、16和50天进行组织取样。" 将在4个体外水平和6个体内水平评估去势对雄激素轴的影响。的 4个水平的体外和体内评估将包括：1）使用qRT-PCR的mRNA水平和使用免疫组织化学（IHC）的蛋白质水平; 2）使用免疫组织化学的雄激素水平。 LC-MS; 3）使用针对PSA、NkxS.1和hK 2的IHC的雄激素调节的基因表达;和4）细胞生长。体内测量将包括5）进展时间和6）存活。ADT后的雄激素代谢将使用雄激素敏感的CWR 22细胞悬液、雄激素依赖的CWR 22异种移植物和新鲜的外科组织异种移植模型。雄激素代谢的变化对ADT后的生存至关重要，在体外使用shRNA或药物靶向，在体内使用慢病毒shRNA和/或药物靶向（目的1）。睾丸通过肾上腺雄激素的分泌内代谢形成的雄激素将使用53-还原酶或使用慢病毒感染递送Sult 2A 1（Aim 2）。最后，将使用慢病毒AR shRNA去除AR 构建体或AR降解小分子（Aim 3）。