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（请参阅说明）： 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来自免疫缺陷的鼠宿主的血清。 最终，这些方法可以应用于患者标本（血清，前列腺活检和骨骼活检），以确定不同雄激素消融疗法在CRPC中的疗效，并有助于确定对这些疗法的抗性机制。雄激素及其前体妊娠的稳定 - 异位稀释液色谱 - 质谱法（LC/MS）将是采用的方法。在此核心中开发和提供的LC/MS分析将是独特的，因为目前没有具有足够敏感性和选择性的现有测定方法来检测和定量所有感兴趣的类固醇。类固醇分析核心的独特资源是访问一组重组酮固醇还原酶（Aldo-Keto还原酶），可用作合成子来生成所需的[13C]标记的内部标准。类固醇分析核心将提供最先进的仪器和方法，这些方法将具有成本效益，可在单个研究人员的实验室中无法常规地测量类固醇水平（雄激素和妊娠）。它将协助研究人员设计实验，以便与可用的分析工具兼容。核心的具体目的如下：[1]提供稳定的同位素稀释LC/MS方法论，用于分析各种生物学基质中的酮类剂（例如，血清，细胞培养，前列腺组织以及前列腺组织和前列腺和骨生物生物）； [2]提供稳定的同位素稀释LC/MS方法，用于分析相同的生物矩阵中的羟基糖基因； [3]提供稳定的同位素稀释LC/MS方法，以分析相同的生物基质中雄激素的妊娠前体。由于这些方法在其他地方无法常规上可用，因此类固醇分析核心将对此P01之外的前列腺癌研究产生影响。