The NF-KappaB-Androgen Receptor Axis Drives Failure of Medical Therapy in Human Benign Prostatic Hyperplasia

NF-KappaB-雄激素受体轴导致人类良性前列腺增生药物治疗失败

• 批准号: 9214498
• 负责人: ROBERT J. MATUSIK
• 金额: $ 42.46万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-16 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9214498
• 关键词: 5 Alpha-Reductase Inhibitor; Accounting; Adrenergic Antagonists; Adrenergic alpha-Antagonists; Age; American; Androgen Receptor; Androgens; Beds; Benign; Benign Prostatic Hypertrophy; Cell Culture Techniques; Cell Line; Cells; Collaborations; DNA Binding; Data; Epithelial; Epithelial Cells; Epithelium; Failure; Feedback; Flow Cytometry; Freezing; Gene Expression; Genes; Growth; Human; In Vitro; Individual; Inflammation; Inflammation Mediators; Length; Ligand Binding Domain; Measurement; Measures; Mediating; Medical; Messenger RNA; Modeling; Molecular; NF-kappa B; Operative Surgical Procedures; Organoids; Oxidoreductase; Pathway interactions; Patients; Pharmaceutical Preparations; Production; Prostate; Prostatic; Protein Isoforms; RNA Splicing; Regulation; Reporting; Resistance; Resources; Route; SRD5A2 gene; Sampling; Severities; Signal Transduction; Stanolone; Steroid Receptors; Steroids; Stress; Symptoms; System; Testing; Testosterone; Time; Tissues; Transcriptional Regulation; Translating; Transrectal Ultrasound; Variant; alpha-adrenergic receptor; associated symptom; biobank; drug testing; insight; liquid chromatography mass spectrometry; lower urinary tract symptoms; new therapeutic target; novel strategies; p65; personalized care; precision medicine; research study; response; self reliance; therapy resistant; three dimensional cell culture; transcriptome sequencing; uptake; urologic

PROJECT SUMMARY With the exception of increasing age and dysregulated androgen activity, little is known on how BPH develops and why patients respond very differently to existing drugs. Steroid 5α-reductase inhibitors (5ARI) block the conversion of testosterone (T) to dihydrotestosterone (DHT), representing a major therapy for shrinking prostate volume in approximately 34% of symptomatic BPH patients. Our data suggest a fundamental mechanism underlying the resistance of BPH to 5ARIs. We show that nuclear factor kappa B (NFκB) mediates induction of Androgen Receptor Variant 7 (AR-V7), an AR isoform that lacks the Ligand Binding Domain (LBD) of the full-length AR (AR-FL), resulting in constitutively active AR signaling even in the absence of DHT. We demonstrate for the first time that severely symptomatic BPH patients express AR-Vs, and that NFκB regulates the expression of AR-FL and AR-V7 mRNA in spontaneously immortalized human benign prostatic cells. Further, expression of NFκB or AR-V7 in benign prostatic cells is sufficient to induce resistance to 5ARI treatment. Also, we show that NFκB and AR-V7 up-regulate steroid-5α-reductase type II (SRD5A2) expression. Consistent with our cell culture experiments, we show that BPH patient samples have elevated levels of NFκB, AR-V7, and SRD5A2 expression. We demonstrate that AR-V7 and SRD5A2 expression significantly correlated with the Transrectal ultrasound (TRUS) prostatic volume and American Urological Association Symptom Scores (AUASS) in BPH patients, measures of the patients' severity of lower urinary tract symptoms (LUTS). This data suggests a BPH model where a loop of self-reliance develops to overcome medical therapy that is triggered by NFκB and AR-V7 to raise SRD5A2 levels and the production of DHT to re- initiate AR-FL activity. Our Hypothesis is that NFκB and AR signaling controls the failed response to 5ARIs in BPH. To unravel the mechanism of failure of 5ARI, we propose three Specific Aims: 1) To determine cross-talk between NFκB and AR signaling to regulate failure of medical therapy; 2) To determine the SRD5A isoforms contribution during resistance to medical therapy; and 3) To determine if failure of medical therapy is driven by NFκB and/or AR-V7 in BPH patients. These aims will use molecular approaches, measurements of androgens and 5ARI levels and gene expression in BPH tissue, and 3D organoid cultures from BPH patients to test the mechanism of failure to 5ARI therapy. Our model may provide a general mechanism explaining failure of medical therapy or it may identify a subpopulation of BPH patients which could point the way towards precision medicine that would personalize care for these patients.

项目摘要 除了年龄增长和雄激素活性失调外，对BPH的发生机制知之甚少 以及为什么病人对现有药物的反应截然不同。类固醇5 α-还原酶抑制剂（5ARI）可阻断 睾酮（T）转化为双氢睾酮（DHT），代表了萎缩的主要疗法 约34%有症状的BPH患者的前列腺体积。我们的数据表明 BPH对5ARI耐药的机制。我们发现，核因子κ B（NF κ B）介导 雄激素受体变体7（AR-V7）的诱导，一种缺乏配体结合结构域（LBD）的AR同种型 的全长AR（AR-FL），导致组成型活跃的AR信号，即使在没有DHT。我们 首次证明了严重症状的BPH患者表达AR-Vs，NF κ B调节 AR-FL和AR-V7 mRNA在自发永生化的人良性前列腺细胞中的表达 此外，良性前列腺细胞中NF κ B或AR-V7的表达足以诱导对5 ARI的抗性 治疗此外，我们发现NF κ B和AR-V7上调类固醇-5 α-还原酶II型（SRD 5A2） 表情与我们的细胞培养实验相一致，我们表明BPH患者样品具有升高的 NF κ B、AR-V7和SRD 5A2表达水平。我们证明了AR-V7和SRD5A2的表达 与经直肠超声（TRUS）前列腺体积和美国泌尿外科 BPH患者的相关症状评分（AUASS），测量患者下尿路疾病的严重程度， 上消化道症状（LUTS）。这些数据表明，BPH模型中，自力更生的循环发展，以克服 通过NF κ B和AR-V7触发的药物治疗，以提高SRD 5A2水平和DHT的产生， 启动AR-FL活动。我们的假设是，NF κ B和AR信号转导控制了对5ARI的失败反应， 前列腺增生。为了揭示5ARI的失效机制，我们提出了三个具体目标：1）确定串扰 NF κ B和AR信号之间的关系，以调节药物治疗失败; 2）确定SRD 5A亚型 药物治疗抵抗期间的贡献;和3）确定药物治疗失败是否由以下因素驱动： NF κ B和/或AR-V7在BPH患者中的表达。这些目标将使用分子方法，测量雄激素 和5ARI水平和基因表达，以及来自BPH患者的3D类器官培养物，以测试 5ARI治疗失败的机制。我们的模型可以提供一个一般的机制，解释失败的 药物治疗，或者它可以识别前列腺增生患者的亚群，这可能为精确治疗指明方向 为这些病人提供个性化护理的药物