ImmunoAnalysis and Research Specimen Management

核心B将为计划项目提供3个主要功能。首先，核心B将向调查人员提供研究标本，并协助鉴定、免疫染色和图像分析。石蜡包埋切片将从原始肿瘤或组织微阵列提供，用于1)使用常规H&E确认组织学；2)使用ACINUS分析细胞凋亡，使用MIB-1检测Ki-67或BrdU掺入的增殖，使用标准的描述性免疫组织化学和视觉评分和/或自动图像分析的定量分析，使用单抗分析雄激素受体；3)应用免疫组织化学和图像分析技术鉴定和定量类固醇受体辅调节因子、雄激素代谢酶、雄激素调节基因产物(PSA、Nkx3.1、α-微管蛋白、翻译延伸因子1a、非神经元烯醇化酶、TomoRegin、硫氧还蛋白还原酶1、MXI-1和人激肽释放酶2)、一般转录因子、生长因子及其受体、干细胞和血管生成生物标志物。第二，Core B将负责临床(冷冻、档案和TMA)和前列腺癌异种移植(CWR22)研究样本的储存、处理和切片。核心人员将采购、处理 并保存去势复发前列腺癌的临床标本，这些标本来自患有晚期前列腺癌的男性，他们因局部复发而出现尿潴留。研究样本将用于构建组织微阵列或激光捕获显微解剖，用于组织雄激素的生化测量。最后，Core B将为该计划项目提供生物统计学专家(Wilding博士)和泌尿生殖系统病理学专家(Alexiev博士)。 Mohler博士和Smith博士将共同指导该设施，并监督已经拥有高水平专业知识并目前为该计划项目履行这些职能的个人的活动。Wilding博士和Alexiev博士将领导将生物统计学和病理学专业知识整合到核心B和3个项目中的努力。核心B发表了8篇方法论文，充分体现了核心B的高水平功能。核心B继续开发更准确地测量蛋白质表达的新方法(4篇出版物)，并在使用TMA进行CAP研究方面保持领先地位(2篇出版物)。核心B为项目1(20)、项目2(8)和项目3(8)提供了出版、印刷和提交手稿的服务。核心B还促进项目之间的互动；23份出版物中有12份涉及一个以上的项目。

Core B will provide 3 primary functions to the Program Project. First, Core B will provide research specimens to the investigators and assist with identification, immunostaining and image analysis. Paraffin-embedded sections will be provided from original tumors or tissue microarrays and used to 1) confirm tissue histology using routine H&E; 2) analyze apoptosis using ACINUS, proliferation using MIB-1 detection of Ki-67 or BrdU incorporation and androgen receptor using monoclonal antibodies by standard, descriptive immunohistochemistry and quantitative analysis using visual scoring and/or automated image analysis; and 3) identify and quantify steroid receptor co-regulators, androgen metabolism enzymes, androgen-regulated gene products (PSA, Nkx3.1, a-tubulin, translation elongation factor 1a, non-neuronal enolase, tomoregulin, thioredoxin reducatase 1, Mxi-1 and human kallikrein 2), general transcription factors, growth factors and their receptors and stem cell and angiogenesis biomarkers using immunohistochemistry and image analysis. Second, Core B will be responsible for storage, processing and sectioning of clinical (frozen, archival and TMAs) and prostate cancer xenograft (CWR22) research specimens. Core personnel will procure, process and store clinical specimens of castration-recurrent prostate cancer obtained from men with advanced prostate cancer who present with urinary retention due to local recurrence. Research specimens will be used to construct tissue microarrays or laser capture microdissected for biochemical measurements of tissue androgens. Lastly, Core B will provide expert biostatistical (Dr. Wilding) and genitourinary pathological service (Dr. Alexiev) to the Program Project. Drs. Mohler and Smith will co-direct the facility and supervise the activities of individuals who already possess high levels of expertise and currently perform these functions for the Program Project. Drs. Wilding and Alexiev will lead the efforts to integrate biostatistical and pathological expertise into Core B and the 3 projects. The high level of function of Core B is demonstrated best by Core B's publication of 8 methods papers. Core B continues to develop new methods for more accurate measurement of protein expression (4 publications) and remains a leader in the use of TMAs for CaP research (2 publications). Core B has performed services that have resulted in manuscripts published, in press and submitted for Project 1 (20), Project 2 (8) and Project 3 (8). Core B also facilitates interaction among the Projects; 12 of 23 publications involved more than 1 Project.