KY COBRE: DISSECTING THE CELL SURFACE PROTEOME OF PROSTATE CANCER

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。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，每年导致大约3万名男性死亡。前列腺癌经常转移到骨骼和淋巴结，导致死亡。对于转移性前列腺癌没有有效的治疗方法。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起着至关重要的作用，如肿瘤细胞脱离细胞外基质、肿瘤细胞抵抗脱落诱导的凋亡、肿瘤细胞粘附内皮细胞和支持肿瘤生长的血管生成。本项目的假设是，在上皮细胞和内皮细胞中存在独特或差异表达的细胞表面蛋白，这些蛋白对前列腺癌转移的个体过程很重要。目前，还没有现成的蛋白质组学方法来表征细胞表面蛋白质组。具体目标1是开发创新的方法来专门表征细胞表面蛋白质组，使用靶向和标记细胞表面蛋白质的新方法和尖端的质谱技术相结合。通过特异性的细胞表面蛋白质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。特异性目的2是使用这些蛋白质组学方法，通过将癌性前列腺上皮细胞与正常前列腺上皮细胞进行比较，来鉴定癌性前列腺上皮细胞中独特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或转染技术抑制或过表达，以确定它们在细胞粘附/脱离和凋亡中的作用。特异性目的3是鉴定前列腺癌经常发生转移的骨髓内皮细胞中独特的细胞表面蛋白。选择其中几种独特的内皮细胞表面蛋白。利用siRNA技术抑制选定蛋白的表达，我们将重点确定它们在癌内皮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参与前列腺癌转移的细胞表面蛋白，并为晚期转移性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.