Steroid Analytical Core

项目总结（见说明）： 类固醇分析核心将为该计划中的所有项目、管理/临床/生物统计核心A和生物标本/动物模型设施核心B提供在不同生物基质中测量雄激素代谢组的复杂分析方法，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细胞系的培养基、去势抵抗性前列腺癌（CRPC）的异种移植物模型和来自其免疫缺陷鼠宿主的血清。 最终，这些方法可以应用于患者标本（血清，前列腺和骨活检），以确定CRPC中不同雄激素消融疗法的疗效，并帮助确定对这些疗法的耐药机制。将采用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）。本核心中开发和提供的LC/MS检测方法将是独特的，因为目前没有具有足够灵敏度和选择性的现有检测方法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获得一组重组酮类固醇还原酶（醛酮还原酶），可用作生成所需[13 C]标记内标物的底物。类固醇分析核心将提供最先进的仪器和方法，这将是具有成本效益的测量类固醇水平（雄激素和孕激素），而不是在个别研究者的实验室常规可用。它将协助调查人员设计实验，使其与现有的分析工具兼容。核心的具体目标如下：[1]提供稳定的同位素稀释LC/MS方法，用于分析各种生物基质中的酮雄激素（例如血清、细胞培养物、前列腺组织以及前列腺和骨活检）; [2]提供稳定同位素稀释LC/MS方法用于分析相同生物基质中的羟基雄激素;和[3]提供稳定的同位素稀释LC/MS方法，用于分析相同生物基质中雄激素的甾烷前体。由于这些方法在其他地方无法常规使用，类固醇分析核心将对前列腺癌研究产生影响。

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.