Molecular Systems Pathology Core

分子系统病理学核心将为该计划项目的研究人员提供人类和小鼠模型的正常和肿瘤样本的专业形态评估。Core将进一步开发组织切片中定量生物标志物多路传输的形态测量和图像分析领域的新技术。该核心还产生了新的分析方法，例如那些用于在高光谱分辨率下使用H2 AX定量分析DNA损伤的方法，用于在微观解剖细节上研究衰老，以及使用新鲜的人类前列腺癌样本进行体外分析。通过这些方法，CORE将与追求系统病理学方法的所有三个项目整合，以调查NKX3.1的表达(CORE的主要科学目标)，研究前列腺干细胞的标记(项目1)，检查NKX3.1的直接转录目标的表达(项目2)，以及量化人类和小鼠前列腺组织中的DNA损伤反应标记(项目3)。我们有充足的组织资源来协助所有项目，包括一组140例人类前列腺癌(训练组)、第二组410例人类前列腺癌(验证组)，以及来自纪念斯隆-凯特琳癌症中心(MSKCC)和西南肿瘤学小组(GU-SWOG)的大量且具有良好特征的人类前列腺癌组织队列。该核心还协助进行免疫组织化学(IHC)和原位杂交(ISH)分析，包括间期荧光原位杂交(FISH)。利用这些资源，我们将表征参与该计划相关途径的已知基因的表达模式，以及通过拟议项目进行的研究确定的新基因。具体目标是：(1)研究前列腺癌前病变和肿瘤病变中与前列腺癌起始相关的分子改变的发生和相关性，这一目标包括技术发展和进一步定义与前列腺癌进展相关的分子改变的时间图谱；(2)在项目1中与Michael Shenin合作，检测NKX3.1和干/祖标记物在雄激素消融和激素完整的人前列腺标本中的表达；(3)使用项目2中确定的正常和肿瘤人标本，与Cory Abate-Shenin合作，表征NKX3.1及其转录靶点的表达；(4)为了进一步研究NKX3失活与TMPRSS2-ERG融合形成的关系，与Edward Gelmann在项目3中对DNA损伤处理前后的小鼠进行了研究。

The Molecular Systems Pathology Core will provide expert morphological evaluation of normal and tumor samples of human and mouse models to investigators of the Program Project. The Core will further develop novel technology in the areas of morphometry and image analysis of quantitative biomarker multiplexing in tissue sections. This Core is also generating new assays, such as those for analysis of DNA damage using H2AX quantitation at high spectral resolution, for studying senescence at the microanatomical detail, and ex vivo assays using fresh human prostate cancer specimens. Through such approaches, the Core will integrate with all three Projects in pursuing systems pathology approaches to investigate expression of NKX3.I (the major scientific goal of the Core), to study markers of prostate stem cells (Project 1), to examine the expression of direct transcriptional targets of NKX3.1 (Project 2), and to quantify DNA damage response markers in human and mouse prostate tissues (Project 3). We have ample tissue resources to assist all Projects, including a set of 140 human prostate tumors (training set), a second independent cohort of 410 human prostate tumors (validation set), as well as large and well characterized cohorts of human prostate cancer tissues from Memorial Sloan-Kettering Cancer Center (MSKCC) and the Genitourinary Program of the Southwestern Oncology Group (GU-SWOG). This Core also provides assistance in conducting immunohistochemical (IHC) and in situ hybridization (ISH) assays, including interphase fluorescence in situ hybridization (FISH). Using these resources, we will characterize expression patterns of known genes participating in pathways of relevance for the Program, as well as novel genes identified through studies conducted by the proposed projects. The specific aims are: (1) To investigate the occurrence and correlation of molecular alterations associated with prostate cancer initiation in human preneoplastic and tumor lesions, an aim that includes both technology development and further definition of the temporal map of molecular alterations associated with prostate cancer progression; (2) To examine the expression of NKX3.1 and stem/progenitor markers in androgen-ablated and hormonally-intact human prostate specimens, working with Michael Shen in Project 1; (3) To characterize the expression of NKX3.1 and its transcriptional targets identified in Project 2 using normal and tumor human specimens, working with Cory Abate-Shen; and (4) To further study the relationship of NKX3.I inactivation and the formation of TMPRSS2-ERG fusions, studying pre- and post-treated mice after DNA damage with Edward Gelmann in Project 3.