RECOMBINANT ANTIBODIES AGAINST PROSTATE CANCER ANTIGENS

抗前列腺癌抗原的重组抗体

• 批准号: 7336080
• 负责人: MYRON NV WILLIAMS
• 金额: $ 11.91万
• 依托单位: CLARK ATLANTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-01 至 2007-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7336080
• 关键词: RECOMBINANT; ANTIBODIES; AGAINST; PROSTATE; CANCER

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. Although an immunochemical test for prostate specific antigen (PSA) test has proved valuable for early diagnosis of prostate adenocarcinoma (PCa), elevated PSA levels are also seen in a number of other prostatic conditions that have distinct therapeutic requirements, such as benign prostate hyperplasia (BPH). Biomarkers other than PSA that can be assessed in biopsy specimens, in serum or in urine will be valuable in determining the appropriate strategy for intervention. Monoclonal antibodies (MAbs) provide a rapid and specific means of detecting the presence of biomarkers, independent of whether they result from changes in gene and protein expression, or from biological modification of proteins or other macromolecules. We propose to isolate antibodies that recognize novel biomarkers associated with tumorigenic potential of prostate cells. The underlying hypothesis of this work is that detectable antigens are differentially expressed in prostate cells as they become oncogenically transformed. Identification of these antigens, some of which are likely to be present on the surfaces of cells, may be performed using paired cell lines that differ in tumorigenic potential. Although MAbs can be easily used to distinguish antigens on cells, isolation of MAbs by traditional, hybridoma methods is difficult in the absence of purified antigen. Antibody phage display permits the use of affinity subtraction and purification techniques for selection of MAbs (Aim 1) binding to antigens that differ between closely related cell types, even without prior knowledge of the nature of the antigens involved. Lineage-related human prostate cell lines, RWPE-1 and RWPE-2, which differ in the absence/presence of a Ki-Ras gene, are an ideal platform for identification of tumorigenicity-associated antigens. Antibody phage display libraries will be subtracted on RWPE-1 and selected on RWPE-2 cell lines. MAbs identified in this manner will be verified for their specificity (Aim 3) for tumorigenic cultured cells, and their cognate antigens characterized (Aim 2). Archived serum, radical prostatectomy and biopsy specimens from localized and metastatic PCa will be probed with the novel MAbs to determine the scope of expression of the antigens in cancer patients. The primary outcome of this study will be novel antibodies against biomarkers associated with tumorigenic prostate disease. This will have an impact on diagnosis and treatment of prostatic cancer in that they will permit the development of diagnostic assays that can predict the tumorous potential of abnormal prostatic growth. Antibodies or other molecules directed against these antigens may also prove novel therapeutic targets and thus have the potential to dramatically reduce mortality.

该子项目是利用 NIH/NCRR 资助的中心拨款提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中出现。列出的机构是中心的机构，不一定是研究者的机构。尽管前列腺特异性抗原 (PSA) 的免疫化学测试已被证明对于前列腺腺癌 (PCa) 的早期诊断有价值，但在许多具有不同治疗要求的其他前列腺疾病中也可以看到 PSA 水平升高，例如良性前列腺增生。前列腺增生）。除 PSA 外，可在活检标本、血清或尿液中进行评估的生物标志物对于确定适当的干预策略非常有价值。单克隆抗体 (MAb) 提供了一种快速、特异的方法来检测生物标志物的存在，无论它们是由基因和蛋白质表达的变化还是由蛋白质或其他大分子的生物修饰引起的。我们建议分离识别与前列腺细胞致瘤潜力相关的新型生物标志物的抗体。这项工作的基本假设是，当前列腺细胞发生致癌转化时，可检测的抗原在前列腺细胞中差异表达。这些抗原（其中一些可能存在于细胞表面）的鉴定可以使用致瘤潜力不同的配对细胞系进行。尽管单克隆抗体可以很容易地用于区分细胞上的抗原，但在没有纯化抗原的情况下，通过传统的杂交瘤方法分离单克隆抗体是困难的。抗体噬菌体展示允许使用亲和力扣除和纯化技术来选择与密切相关的细胞类型之间不同的抗原结合的 MAb（目标 1），即使事先不了解所涉及抗原的性质。谱系相关的人类前列腺细胞系 RWPE-1 和 RWPE-2 的不同之处在于是否存在 Ki-Ras 基因，是鉴定致瘤性相关抗原的理想平台。抗体噬菌体展示文库将在 RWPE-1 上扣除，并在 RWPE-2 细胞系上选择。以这种方式鉴定的单克隆抗体将验证其对致瘤培养细胞的特异性（目标 3），并鉴定其同源抗原（目标 2）。存档的血清、根治性前列腺切除术以及局部和转移性 PCa 的活检标本将用新型 MAb 进行探测，以确定癌症患者中抗原的表达范围。这项研究的主要成果将是针对与致瘤性前列腺疾病相关的生物标志物的新型抗体。这将对前列腺癌的诊断和治疗产生影响，因为它们将允许开发能够预测异常前列腺生长的肿瘤潜力的诊断测定法。针对这些抗原的抗体或其他分子也可能被证明是新的治疗靶点，因此有可能显着降低死亡率。