Genomic and proteomic analysis of prostate cancer

描述（由申请人提供）：局部前列腺癌可通过手术或放疗治疗。然而，很大比例的男性在最初诊断时已经有转移性疾病。转移性前列腺癌唯一有效的全身疗法是雄激素剥夺。雄激素剥夺不能完全和永久地消除所有前列腺癌细胞群，表现为可预测的初始反应和复发模式，最终进展为雄激素独立。雄激素剥夺与前列腺癌细胞通过雄激素依赖、雄激素敏感和最终雄激素独立的光谱逐渐过渡有关。越来越多的证据支持这样一种观点，即前列腺癌的进展伴随着依赖内分泌控制向旁分泌控制的转变，最终是自分泌控制的转变，这一复杂的过程是发生在细胞控制分子水平上的变化的结果。然而，雄激素非依赖性前列腺癌发生的分子机制尚不清楚。由于活检材料的细胞异质性和缺乏理想的体内模型，这些分子机制的研究一直受到阻碍。现有的人类异种移植物模型在阉割宿主后发展为雄激素独立，产生的肿瘤被宿主细胞高度污染。因此，我们开发了一种体内模型，包括使用中空纤维回收未受污染的前列腺癌细胞（肿瘤）包，可用于前列腺癌进展到雄激素独立的后续分子生物学分析。我们建议在雄激素非依赖性肿瘤进展开始之前和之后从动物身上收集的细胞中表征基因表达。特异性Aim 1将采用基因表达技术（SAGE）和Affymetrix GeneChips的序列分析，而在Aim 2中，将使用ICATLC/ MC/MS和二维凝胶电泳（2D gel）来鉴定前列腺癌细胞向雄激素独立发展过程中蛋白质表达的变化。需要双管齐下的方法来确定在雄激素独立的过程中转录组和蛋白质组表达的全局变化。目的1和2的结果将通过原位杂交和/或免疫组织化学对前列腺癌患者的临床样本进行原位杂交和异种移植到宿主前后的验证。我们处于一个独特的位置，在前列腺癌细胞发展到雄激素独立的阶段，我们可以获得唯一的模型，提供未受宿主细胞污染的RNA和蛋白质来源，经过验证的专业知识和设施，用于SAGE文库建设，测序，生物信息学分析，以及一种新的人类异种移植模型。从这些研究中获得的数据将用于确定前列腺癌向雄激素独立发展的重要途径和分子机制。只有通过识别这些途径和机制，才能开发新的靶点和治疗方法，从而可能延缓或避免前列腺癌向雄激素不依赖型疾病的发展。

DESCRIPTION (provided by applicant): Localized prostate cancer can be treated surgically or by radiotherapy. However, a large percentage of men will already have metastatic disease upon initial diagnosis. The only effective systemic therapy available for metastatic prostate cancer is androgen deprivation. The inability of androgen deprivation to completely and permanently eliminate all prostate cancer cell populations is manifested by the predictable pattern of initial response and relapse with the ultimate progression to androgen independence. Androgen deprivation is associated with a gradual transition of prostate cancer cells through a spectrum of androgen dependence, androgen sensitivity and ultimately androgen independence. There is mounting evidence supporting the concept that prostate cancer progression is accompanied by a shift in reliance on endocrine controls to paracrine and eventually autocrine controls and that this complex process is the result of changes, which occur at molecular levels of cellular control. However, the molecular mechanisms involved in the development of androgen independent prostate cancer are unknown. Investigation of these molecular mechanisms has been impeded by problems related to cell heterogeneity of biopsy material and the lack of an ideal in vivo model. Available human xenograft models that progress to androgen independence after castration of the host yield tumors that are highly contaminated with host cells. Therefore, we have developed an in vivo model that encompasses the use of hollow fibers to retrieve uncontaminated packages of prostate cancer cells (tumors) that can be used for subsequent molecular biology analyses of the progression of prostate cancer to androgen independence. We propose to characterize gene expression in cells harvested from animals both prior and subsequent to the onset of androgen independent tumor progression. Specific Aim 1 will employ serial analysis of gene expression technology (SAGE) and Affymetrix GeneChips, while in Aim 2, ICATLC/ MC/MS and two-dimensional gel electrophoresis (2D gels) will be used to identify changes in protein expression in during progression of prostate cancer cells to androgen independence. A double pronged approach is required to identify global changes in expression in the transcriptome and proteome during progression to androgen independence. The results of Aims 1 and 2 will be confirmed using in situ hybridization and/or immunohistochemistry in clinical samples from prostate cancer patients before and after xenografting into hosts. We are in a unique position, having both access to the only model available that provides uncontaminated (by host cells) sources of RNA and protein during the stages of progression of prostate cancer cells to androgen independence, proven expertise and facilities for SAGE library construction, sequencing, bioinformatic analysis, and a novel human xenograft model. The data obtained from these studies will be used to identify important pathways and molecular mechanisms involved in the progression of prostate cancer to androgen independence. Only through the identification of these pathways and mechanisms can new targets and therapeutics be developed that may potentially delay or avert the progression of prostate cancer to androgen independent disease.