Genomic and proteomic analysis of prostate cancer

前列腺癌的基因组和蛋白质组分析

• 批准号: 6719486
• 负责人: MARIANNE D SADAR
• 金额: $ 22.14万
• 依托单位: BRITISH COLUMBIA CANCER AGENCY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6719486
• 关键词: SCID mouse; affinity labeling; androgens; athymic mouse; bioinformatics; biotechnology; cell growth regulation; clinical research; electrospray ionization mass spectrometry; gene expression; hormone related neoplasm /cancer; human subject; immunocytochemistry; in situ hybridization; liquid chromatography mass spectrometry; messenger RNA; metastasis; microarray technology; neoplasm /cancer genetics; neoplastic growth; neoplastic process; posttranslational modifications; prostate neoplasms; proteomics; serial analysis of gene expression; two dimensional gel electrophoresis

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.

描述（由申请人提供）：局限性前列腺癌可以通过手术或放射疗法治疗。然而，很大一部分男性在初步诊断时已经患有转移性疾病。对于转移性前列腺癌唯一有效的全身疗法是雄激素剥夺。雄激素剥夺无法完全和永久消除所有前列腺癌细胞群，其表现是可预测的初始反应和复发以及最终进展为雄激素独立的模式。雄激素剥夺与前列腺癌细胞通过一系列雄激素依赖性、雄激素敏感性和最终的雄激素独立性的逐渐转变有关。越来越多的证据支持这样的概念，即前列腺癌的进展伴随着对内分泌控制的依赖向旁分泌控制和最终自分泌控制的转变，并且这一复杂的过程是发生在细胞控制的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。由于与活检材料的细胞异质性相关的问题和缺乏理想的体内模型，这些分子机制的研究受到阻碍。现有的人类异种移植模型在宿主去势后进展到雄激素独立性，产生被宿主细胞高度污染的肿瘤。因此，我们开发了一种体内模型，其中包括使用中空纤维来回收未受污染的前列腺癌细胞（肿瘤）包，该模型可用于前列腺癌向雄激素独立性进展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发生之前和之后表征从动物收获的细胞中的基因表达。具体目标 1 将采用基因表达技术 (SAGE) 和 Affymetrix GeneChips 的系列分析，而在目标 2 中，将使用 ICATLC/MC/MS 和二维凝胶电泳 (2D 凝胶) 来识别进展过程中蛋白质表达的变化。前列腺癌细胞的雄激素独立性。需要双管齐下的方法来识别在雄激素独立过程中转录组和蛋白质组表达的整体变化。目标 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异种移植到宿主之前和之后的临床样本进行证实。我们处于独特的地位，可以使用唯一可用的模型，在前列腺癌细胞发展到雄激素独立的阶段提供未受宿主细胞污染的 RNA 和蛋白质来源，以及用于 SAGE 文库构建的经过验证的专业知识和设施，测序、生物信息分析和新型人类异种移植模型。从这些研究中获得的数据将用于确定前列腺癌进展为雄激素独立性的重要途径和分子机制。只有通过识别这些途径和机制，才能开发出可能延迟或避免前列腺癌进展为雄激素非依赖性疾病的新靶点和治疗方法。