Human aldo-keto reductases and nuclear receptor action

人醛酮还原酶和核受体作用

基本信息

批准号：
7824959
负责人：
Trevor M Penning
金额：
$ 50.58万
依托单位：
UNIVERSITY OF PENNSYLVANIA
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 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7824959
关键词：
2,4-thiazolidinedione 9-deoxy-delta-9-prostaglandin D2 AKR1C1 Ablation Accounting Acetates Address Adenocarcinoma Adrenal Glands Affect Age Agonist American Anabolism Androgen Metabolism Androgen Receptor Androgens Androstenedione Anti-Inflammatory Agents Anti-inflammatory Antineoplastic Agents Apoptosis Area Attenuated Bicalutamide Biological Assay Breast Breast Cancer Cell COS Cells COS-1 Cells CYP17A1 gene Cancer Etiology Cancer Patient Cancer cell line Castration Cathepsins Cell model Cells Cessation of life Chemopreventive Agent Chloride Ion Chlorides Cholesterol Coupled Data Detection Diagnosis Dinoprost Disease Due Process Economics Electrons Enzymes Equilibrium Estradiol Estrogen Receptors Estrogens Estrone Family Funding G alpha q Protein Gene Expression Gene Expression Profile Genes Gland Gleason Grade for Prostate Cancer Goals Grant Growth HSD17B2 gene Hormone Responsive Hormones Human Hydroxysteroid Dehydrogenases Immunohistochemistry Institution Ketoconazole LNCaP Lasers Lead Ligands Liquid Chromatography Location Lyase MCF7 cell Malignant Neoplasms Malignant neoplasm of prostate Mass Spectrum Analysis Measurement Measures Medicine Mefenamic Acid Metabolic Metabolism Methodology Methods Normal tissue morphology Nuclear Hormone Receptors Nuclear Receptors Occupations Operative Surgical Procedures Outcome Oxidoreductase PGF receptor PTEN gene PTGS2 gene Pathway interactions Patients Pattern Peroxisome Proliferators Pharmaceutical Preparations Play Positioning Attribute Pregnenolone Prior Therapy Procedures Production Progesterone Receptors Property Prostaglandin D2 Prostaglandins Prostaglandins F Prostate Prostatectomy Prostatic Prostatic Neoplasms Protein Isoforms RNA Radical Prostatectomy Reaction Receptor Signaling Recovery Refractory Regulation Relapse Reporter Genes Research Resistance Role SRD5A2 gene Sampling Series Signal Transduction Somatotropin Specimen Staining method Stains Steroids Subgroup Testing Testosterone Thiazolidinediones Time Training Transcript Up-Regulation abiraterone analog anticancer research base cell growth cell type cyclooxygenase 1 dehydroepiandrosterone deprivation diabetic enzyme structure fenamic acid inhibitor/antagonist laser capture microdissection liquid chromatography mass spectrometry male malignant breast neoplasm man member molecular pathology multiple reaction monitoring parent grant programs prostaglandin-F synthase public health relevance radiotracer research study response stable isotope steroid hormone

2,4-thiazolidinedione 9-deoxy-delta-9-prostaglandin D2 AKR1C1 Ablation Accounting Acetates Address Adenocarcinoma Adrenal Glands Affect Age Agonist American Anabolism Androgen Metabolism Androgen Receptor Androgens Androstenedione Anti-Inflammatory Agents Anti-inflammatory Antineoplastic Agents Apoptosis Area Attenuated Bicalutamide Biological Assay Breast Breast Cancer Cell COS Cells COS-1 Cells CYP17A1 gene Cancer Etiology Cancer Patient Cancer cell line Castration Cathepsins Cell model Cells Cessation of life Chemopreventive Agent Chloride Ion Chlorides Cholesterol Coupled Data Detection Diagnosis Dinoprost Disease Due Process Economics Electrons Enzymes Equilibrium Estradiol Estrogen Receptors Estrogens Estrone Family Funding G alpha q Protein Gene Expression Gene Expression Profile Genes Gland Gleason Grade for Prostate Cancer Goals Grant Growth HSD17B2 gene Hormone Responsive Hormones Human Hydroxysteroid Dehydrogenases Immunohistochemistry Institution Ketoconazole LNCaP Lasers Lead Ligands Liquid Chromatography Location Lyase MCF7 cell Malignant Neoplasms Malignant neoplasm of prostate Mass Spectrum Analysis Measurement Measures Medicine Mefenamic Acid Metabolic Metabolism Methodology Methods Normal tissue morphology Nuclear Hormone Receptors Nuclear Receptors Occupations Operative Surgical Procedures Outcome Oxidoreductase PGF receptor PTEN gene PTGS2 gene Pathway interactions Patients Pattern Peroxisome Proliferators Pharmaceutical Preparations Play Positioning Attribute Pregnenolone Prior Therapy Procedures Production Progesterone Receptors Property Prostaglandin D2 Prostaglandins Prostaglandins F Prostate Prostatectomy Prostatic Prostatic Neoplasms Protein Isoforms RNA Radical Prostatectomy Reaction Receptor Signaling Recovery Refractory Regulation Relapse Reporter Genes Research Resistance Role SRD5A2 gene Sampling Series Signal Transduction Somatotropin Specimen Staining method Stains Steroids Subgroup Testing Testosterone Thiazolidinediones Time Training Transcript Up-Regulation abiraterone analog anticancer research base cell growth cell type cyclooxygenase 1 dehydroepiandrosterone deprivation diabetic enzyme structure fenamic acid inhibitor/antagonist laser capture microdissection liquid chromatography mass spectrometry male malignant breast neoplasm man member molecular pathology multiple reaction monitoring parent grant programs prostaglandin-F synthase public health relevance radiotracer research study response stable isotope steroid hormone

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项目摘要

DESCRIPTION (provided by applicant): Penn Medicine contributes substantially to the local economy. In 2008, Penn Medicine created 37,000 jobs and $5.4 billion in regional economic activity, with the area's highly trained workforce producing more than 24,600 applications for just 840 open Penn staff research positions. The current proposal will create or retain 5 jobs at two different institutions. This is an application for a Competitive Revision to 1R01-CA90744: "Aldo-Keto Reductases and Nuclear Receptor Action" (Project End Date: 06/30/12) to be funded by the American Recovery and Reinvestment Act of 2009. The parent grant is focused on the role of AKR1C3 (type 5 17¿-hydroxysteroid dehydrogenase (17¿-HSD) or prostaglandin F synthase) in regulating the local production of ligands for nuclear receptors in prostate and breast cancer. Our hypothesis is that prostate cancer is a disease of the aging male and that it is driven by the local production of androgens in the gland rather than by androgens of testicular origin. The precursor for this intracrine formation of androgens is likely dehydroepiandrosterone of adrenal origin or pregnenolone of prostatic origin. AKR1C3 plays a key role in prostate androgen biosynthesis since it converts ?4-androstene-3,17-dione (a weak androgen) to testosterone (a potent androgen) and lies immediately upstream from type 2 5a-reductase. Recently, castration resistant prostate cancer has been shown to undergo a re-programming of androgen biosynthesis due to androgen deprivation. Part of this response includes upregulation of several genes involved in androgen biosynthesis including AKR1C3. Moreover, abiraterone acetate a CYP17,20-lyase inhibitor has been found effective in advanced prostate cancer suggesting that local androgen signaling may still be occurring. To test this hypothesis it is critical to have reliable methods to measure intraprostatic androgen levels in different patient groups and relate them to enzyme expression levels. This supplement will therefore go beyond the initial funded specific aims of the grant to: [1] Develop a stable-isotope dilution LC/MS assay for the detection and quantitation of all androgen metabolites downstream from pregnenolone and dehydroepiandrosterone (DHEA); [2] Validate the method by measuring androgen metabolic profiles in LNCaP and LNCaP-AKR1C3 stably transfected cells; [3] Apply the method to measure intrapostatic androgen levels in adenocarcinoma of patients that have undergone radical prostatectomy (positive control) and adjacent normal tissue; and in prostatectomy samples in patients that underwent different androgen deprivation therapies prior to surgery, e.g. LHRH agonist alone; LHRH agonist + bicalutamide (androgen receptor antagonist); and LHRH agonist + bicalutamide + ketoconazole (a CYP17,20- lyase inhibitor); and [4] Use qPCR to measure the expression of all steroidogenic enzymes after cholesterol in the same prostate specimens: Scc, CYP17A1, HSD3B2, AKR1C3, SRD5A1, SRD5A2, AKR1C2, AKR1C1, HSD17B2, HSD17B6, and UGT2B15 to determine if their levels correlate with the androgen levels observed. These studies will lead to a robust method that can measure intraprostatic androgen levels in prostate cancer patients that have undergone radical prostatectomy and determine the efficacy of androgen deprivation therapy. They will also determine whether prostatic androgen synthesis can occur from pregnenolone and whether elevated androgen levels can be correlated to increased expression of steroidogenic enzymes including AKR1C3. PUBLIC HEALTH RELEVANCE: Prostate cancer is the second leading cause of cancer death in man. It is initially responsive to androgen ablative therapy but thereafter patients relapse and develop castration resistant prostate cancer (CRPC). Emerging data suggests that CRPC has undergone a re-programming or adaptive response to androgen ablation and synthesizes its own androgens. Aldo-keto reductase (AKR) family 1C member 3 (AKR1C3) also known as type 5 17¿-hydroxysteroid dehydrogenase may play a key role in this response. This proposal is aimed at developing a stable isotope dilution liquid chromatography mass spectrometry method to measure the intraprostatic androgen metabolome which could be used to better diagnose and treat prostate cancer.

描述（由应用程序提供）：宾夕法尼亚医学对当地经济做出了重大贡献。 2008年，宾夕法尼亚医学公司创造了37,000个工作岗位和54亿美元的区域经济活动，该地区训练有素的劳动力生产超过24,600份申请，仅用于840个开放式宾夕法尼亚州工作人员研究职位。当前的提案将在两个不同机构创建或保留5个工作。这是对1R01-CA90744进行竞争性修订的申请：“ Aldo-Keto还原酶和核受体动作”（项目结束日期：06/30/12）由2009年的《美国恢复和再投资法》提供资金。父母赠款的重点是AKR1C3（57型dehdredroxystrase of typlags in typlogn of defragn foragtion for）合成酶）在预定前列腺核受体和乳腺核受体的局部生产时，我们的假设是前列腺癌是衰老雄性的疾病，它是由腺体中雄激素的局部产生驱动的，而不是由雄激素驱动。这种雄激素内形成的前体可能是肾上腺肾上腺雌激素的肾上腺素或前列腺起源的untixtolone。 AKR1C3在前列腺雄激素生物合成中起关键作用，因为它转化了4- androstene-3,17-二酮（一种弱雄激素），向睾丸激素（潜在的雄激素），并立即从2型5A-还原酶上游放置。最近，已证明耐castration抗性前列腺癌会因雄激素剥夺而进行雄激素生物合成的重新编程。该响应的一部分包括上调包括AKR1C3在内的雄激素生物合成的几个基因。此外，已经发现Abiraterone乙酸CYP17,20-裂解剂在晚期前列腺癌中有效，这表明仍可能发生局部雄激素信号传导。为了检验这一假设，至关重要的是，具有可靠的方法来测量不同患者群体中肉眼内雄激素水平并将其与酶表达水平联系起来。因此，该补充剂将超越赠款的最初资助的特定目的：[1]开发一种稳定的同位素稀释LC/MS测定法，以检测和定量来自Timhtolone和脱氢表雄酮（DHEA）下游的所有雄激素代谢物； [2]通过测量LNCAP和LNCAP-AKR1C3稳定翻译细胞中的雄激素代谢谱来验证该方法； [3]应用方法测量具有根本根治性前列腺切除术（阳性对照）和相邻正常组织的患者腺癌中的静脉内雄激素水平；以及在手术前接受不同雄激素剥夺疗法的患者的前列腺切除术样品中，例如单独使用LHRH激动剂； [4]使用QPCR在同一前列腺样品中测量胆固醇后所有类固醇生成酶的表达：SCC，CYP17A1，HSD3B2，AKR1C3，SRD5A1，SRD5A1，SRD5A2，SRD5A2，SRD5A2，AKR1C2，AKR1C2，AKR1C1，AKR1C1，HSD17B217B2，HSD17B617B617B6，和HSD17B6，和HSD17B6，and217b6，和HSD17B6，and217b6，and2。与观察到的雄激素水平相关。这些研究将导致一种可靠的方法，该方法可以在前列腺癌患者中测量肉眼内雄激素水平，这些患者接受了根治性的前列腺切除术并确定雄激素剥夺疗法的效率。他们还将确定是否可以从unteryolone中发生前列腺雄激素合成，以及是否可以将升高的雄激素水平与包括AKR1C3在内的类固醇生成酶的表达增加相关。公共卫生相关性：前列腺癌是人类癌症死亡的第二大原因。它最初对雄激素消融疗法有反应，但此后患者中继并发展为抑制castration抗性前列腺癌（CRPC）。新兴数据表明，CRPC对雄激素消融的重新编程或适应性反应进行了反应，并综合了其自己的雄激素。 Aldo -Keto还原酶（AKR）家族1C成员3（AKR1C3）也称为5型17？-Hydroxyteroid脱氢酶可能在此反应中起关键作用。该建议旨在开发一种稳定的同位素稀释液色谱质谱法，以测量肉眼内雄激素代谢组，可用于更好地诊断和治疗前列腺癌。