KY COBRE: DISSECTING THE CELL SURFACE PROTEOME OF PROSTATE CANCER

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。前列腺癌是美国所有年龄段男性癌症相关死亡的主要原因，每年导致约30,000名男性死亡。前列腺癌经常转移到骨骼和淋巴结，导致死亡。目前还没有治疗转移性前列腺癌的有效方法。该项目的长期目标是了解细胞表面蛋白在前列腺癌转移中的作用。细胞表面蛋白在前列腺癌转移的多个步骤中发挥重要作用，如肿瘤细胞与细胞外基质的分离、肿瘤细胞对分离诱导的细胞凋亡的抵抗、肿瘤细胞与内皮细胞的黏附以及支持肿瘤生长的血管生成。这个项目的假设是，在上皮细胞和内皮细胞中都有独特或差异表达的细胞表面蛋白，这些蛋白在前列腺癌转移的个别过程中起重要作用。目前，还没有蛋白质组学方法可以很容易地描述细胞表面蛋白质组。具体目标1是开发创新的方法，利用靶向和标记细胞表面蛋白的新方法和尖端的质谱学技术相结合，专门表征细胞表面蛋白质组。通过特定的细胞表面蛋白靶向和高精度的蛋白质鉴定和定量，将实现对细胞表面蛋白质组的全面了解。具体目的2是使用这些蛋白质组学方法，通过将其与正常前列腺上皮细胞进行比较，来鉴定癌前列腺上皮细胞中唯一或差异表达的细胞表面蛋白。将使用多种策略来选择有限数量的蛋白质，这些蛋白质将成为功能研究的重点。选择的癌症相关蛋白将分别使用siRNA或转基因技术进行抑制或过表达，以确定它们在细胞黏附/脱离和细胞凋亡中的作用。具体目标3是从前列腺癌转移频繁发生的骨髓中鉴定内皮细胞中独特的细胞表面蛋白。这些独特的内皮细胞表面蛋白将被挑选出来。利用siRNA技术抑制所选蛋白的表达，我们将重点研究它们在肿瘤-内皮细胞-细胞相互作用和血管生成中的功能作用。本文提出的研究将确定一系列可能与前列腺癌转移有关的细胞表面蛋白，并为晚期转移疾病的治疗干预开辟新的途径。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Prostate cancer is a major contributor to cancer related mortality in American men of all ages, causing the death of approximately 30,000 men each year. Prostate cancer frequently metastasizes to the bone and lymph nodes, resulting in mortality. No effective treatment for metastatic prostate cancer is available. The long-term objective of this project is to understand the role of cell surface proteins in prostate cancer metastasis. Cell surface proteins play vital roles in multiple steps of prostate cancer metastasis, such as detachment of tumor cells from the extracellular matrix, resistance of tumor cells to the detachment-induced apoptosis, adhesion of tumor cells to endothelial cells and angiogenesis to support tumor growth. The hypothesis of this project is that there are cell surface proteins either uniquely or differentially expressed in both epithelial and endothelial cells that are important for individual processes of prostate cancer metastasis. At present, no proteomic approaches are readily available to characterize the cell surface proteome. Specific Aim 1 is to develop innovative approaches to specially characterize cell surface proteomes using a combination of new methods targeting and labeling cell surface proteins and cutting-edge mass spectrometry technologies. A comprehensive understanding of the cell surface proteomes will be achieved by specific cell surface protein targeting and high accuracy protein identification and quantification. Specific Aim 2 is to use these proteomic approaches to identify uniquely or differentially expressed cell surface proteins in cancerous prostate epithelial cells by comparing them to normal prostate epithelial cells. Multiple strategies will be used to select a limited number of proteins that will be the focus of functional studies. The selected cancer-associated proteins will be either suppressed or overexpressed using siRNA or transfection techniques respectively to determine their role in cell adhesion/detachment and apoptosis. Specific Aim 3 is to identify unique cell surface proteins in endothelial cells from bone marrow where prostate cancer metastasis frequently occurs. Several those unique endothelial cell surface proteins will be selected. Using the siRNA technique to suppress the expression of the selected proteins, we will focus on determining their functional roles in carcinoma-endothelial cell-cell interaction and angiogenesis. The studies proposed herein will determine an array of cell surface proteins potentially involved in prostate cancer metastasis and open new avenues for therapeutic intervention of the advanced metastatic disease.