Steroid Analytical Core

类固醇分析核心

• 批准号: 8475916
• 负责人: Trevor M Penning
• 金额: $ 30.87万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-24 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8475916
• 关键词: Alternative Splicing; Anabolism; Androgen Metabolism; Androgen Receptor; Androgens; Animal Model; Biological; Biological Assay; Biopsy; Biostatistics Core; Castration; Cell Culture Techniques; Cholesterol; Clinical; Core Facility; Development; Drug resistance; EZH2 gene; Epigenetic Process; Genes; Goals; Health; Histone H3; Individual; Instruction; Ketosteroids; Label; Laboratories; Lead; Ligands; Malignant neoplasm of prostate; Measurement; Measures; Methodology; Methods; Mus; Outcome; Oxidoreductase; PC3 cell line; Patients; Performance; Pregnanes; Prostate; RNA Splicing; Recombinants; Research Personnel; Research Project Grants; Resistance; Resources; Sampling; Serum; Specimen; Steroids; Tissues; Transferase; Variant; Xenograft Model; analytical tool; anticancer research; base; bone; castration resistant prostate cancer; cost effective; deprivation; design; instrumentation; interest; liquid chromatography mass spectrometry; programs; receptor function; research study; resistance mechanism; stable isotope; steroid metabolism; therapy resistant

PROJECT SUMMARY (See instructions): The Steroid Analytical Core will provide sophisticated analytical methodologies for measuring the androgen metabolome with sufficient sensitivity and selectivity in different biological matrices for all Projects in the Program, for the Administrative/Clinical/Biostatistics Core A and for the Biospecimen/Animal Model Facility Core B. These matrices include medium from prostate cancer lines maintained in culture, xenograft models of castrate resistant prostate cancer (CRPC), and serum from their immunodeficient murine hosts. Ultimately, these methods can be applied to patient specimens (serum, prostate and bone biopsies) to determine the efficacy of different androgen ablative therapies in CRPC, and to help identify mechanisms of resistance to these therapies. Stable-isotope dilution liquid chromatography-mass spectrometry (LC/MS) of androgens and their precursor pregnanes will be the methodology employed. The LC/MS assays developed and provided in this core will be unique since no existing assays with sufficient sensitivity and selectivity are currently available to detect and quantitate all the steroids of interest. A unique resource of the Steroid Analytical Core is access to a panel of recombinant ketosteroid reductases (aldo-keto reductases) that can be used as synthons to generate the required [13C]-labeled internal standards. The Steroid Analytical Core will provide access to state-of-the art instrumentation and methods that will be cost-effective to measure steroid levels (androgens and pregnanes) not routinely available in the laboratories of individual investigators. It will assist investigators in the design of their experiments so that they are compatible with the analytical tools available. The specific aims of the Core are as follows: [1] To provide stable isotope dilution LC/MS methodology for the analysis of ketoandrogens in a variety of biological matrices (e.g. serum, cell culture, prostate tissue, and prostate and bone biopsies); [2] To provide stable isotope dilution LC/MS methodology for the analysis of hydroxyandrogens in identical biological matrices; and [3] To provide stable isotope dilution LC/MS methodology for the analysis of pregnane precursors of androgens in identical biological matrices. Since these methods are not routinely available elsewhere the Steroid Analytical Core will have an impact on prostate cancer research beyond this P01.

项目摘要（参见说明）： 类固醇分析核心将为该计划中的所有项目、行政/临床/生物统计核心 A 和生物样本/动物模型设施核心 B 提供复杂的分析方法，用于测量不同生物基质中的雄激素代谢组，具有足够的灵敏度和选择性。这些基质包括来自培养物中维持的前列腺癌系的培养基、去势抵抗性前列腺癌（CRPC）的异种移植模型以及来自其免疫缺陷小鼠宿主的血清。 最终，这些方法可以应用于患者样本（血清、前列腺和骨活检），以确定不同雄激素消融疗法对 CRPC 的疗效，并帮助确定对这些疗法的耐药机制。采用的方法将是雄激素及其前体孕烷的稳定同位素稀释液相色谱-质谱法 (LC/MS)。该核心开发和提供的 LC/MS 测定将是独一无二的，因为目前没有具有足够灵敏度和选择性的现有测定可用于检测和定量所有感兴趣的类固醇。类固醇分析核心的一个独特资源是访问一组重组酮类固醇还原酶（醛酮还原酶），这些酶可用作合成子来生成所需的 [13C] 标记内标。类固醇分析核心将提供最先进的仪器和方法，这些仪器和方法将经济高效地测量类固醇水平（雄激素和孕烷），而这些在个别研究人员的实验室中是常规无法获得的。它将帮助研究人员设计实验，使其与可用的分析工具兼容。核心的具体目标如下： [1] 提供稳定同位素稀释 LC/MS 方法来分析各种生物基质（例如血清、细胞培养物、前列腺组织以及前列腺和骨活检）中的酮雄激素； [2] 为分析相同生物基质中的羟基雄激素提供稳定同位素稀释液质联用方法； [3] 提供稳定同位素稀释 LC/MS 方法来分析相同生物基质中雄激素的孕烷前体。由于这些方法在其他地方通常不可用，类固醇分析核心将对前列腺癌研究产生超出 P01 的影响。