Synthesis of MUC1 glycopeptides and generation of antibodies for the indentificat

说明书(申请人提供)：MUC1糖肽的合成和用于鉴定癌细胞表位的抗体的产生。大多数癌症过度表达细胞表面糖蛋白，糖蛋白被截短的O-糖链异常糖基化，O-糖链是肿瘤相关抗原。健康细胞和癌细胞在细胞表面组成上的明显不同使肿瘤相关抗原成为癌症特异性免疫疗法发展的靶点。最近，人们发现糖和多肽结合的表位是癌症免疫治疗的极佳靶点。在所有异常糖基化的癌细胞表面糖蛋白中，研究最多的是粘蛋白MUC1，然而，由癌细胞动态表达的免疫原性MUC1糖肽的结构特征仍未得到充分研究。目前还没有工具来分析表位的糖部分和肽部分。为了解决这个问题，这里提出的研究结合了候选糖肽的化学合成(包含14个TN或STN的糖肽的文库)和产生的单抗，这将在以后用于特异性地识别癌细胞表面表位(长期目标)。此外，这些探针将被用来研究肿瘤进展过程中表位表达的变化。该SC2试验项目的目标是合成MUC1糖肽文库，产生第一组单抗探针，并验证它们与它们所诱导的糖肽的结合特异性。获得一套全面的此类单抗将对开发新的癌症特异性免疫疗法、诊断和疫苗具有重大意义。所有糖肽都将在德克萨斯大学埃尔帕索分校的P.I.S实验室合成。P.I.S实验室还将在德克萨斯大学埃尔帕索分校的边界生物医学核心设施测定针对MUC1源性糖肽的单抗的结合亲和力和结合特异性。将与德克萨斯州休斯敦大学安德森癌症中心黑色素瘤和内科医学博士周大鹏合作，对小鼠进行免疫接种，并培养出单抗。 公共卫生相关性：MUC1是一种细胞表面糖蛋白，在许多癌症中过度表达和异常糖基化。存在于细胞表面的肿瘤相关抗原是开发有效免疫治疗的理想靶点，然而，肿瘤细胞表达的表位的确切结构特征尚不清楚。我们的目标是开发一套能够特异性识别这些细胞表面糖肽表位的工具--一组单抗。

DESCRIPTION (provided by applicant): Synthesis of MUC1 glycopeptides and generation of antibodies for the identification of epitopes on cancer cells. Most carcinomas over-express cell surface glycoproteins that are aberrantly glycosylated with truncated O-glycans, which are tumor-associated antigens. This distinct difference in cell surface make-up of healthy and cancerous cells make tumor- associated antigens attractive targets for the development of cancer-specific immunotherapies. Recently, it was discovered that combined sugar and peptide epitopes make excellent targets for immunotherapy of cancer. Of all aberrantly glycosylated cancer cell surface glycoproteins the one best studied is the mucin "MUC1", however, the structural features of immunogenic MUC1 glycopeptides, dynamically expressed by cancer cells, remain understudied. Tools to analyze the sugar part and peptide part of an epitope are unavailable. In order to address this question the research proposed here combines the chemical synthesis of candidate glycopeptides (a library of 14 Tn or STn containing glycopeptides), and the generation of monoclonal antibodies, which will later be used to specifically identify cancer cell surface epitopes (long-term goal). Furthermore, these probes will be used to study changes in epitope expression as the cancer progresses. The objective of this SC2 pilot project is to synthesize a MUC1 glycopeptide library, to generate a first set of monoclonal antibody probes, and to validate their binding specificity to the glycopeptides they have been elicited against. The availability of a comprehensive set of such monoclonal antibodies would have major implications for the development of novel cancer-specific immunotherapies, diagnostics and vaccines. All glycopeptides will be synthesized in the P.I.'s laboratory at the University of Texas at El Paso. The P.I.'s laboratory will also determine the binding affinities and binding specificities of the monoclonal antibodies to the MUC1 derived glycopeptides at the Border Biomedical Core Facility at the University of Texas at El Paso. Mice will be immunized and monoclonal antibodies will be raised in collaboration with Dapeng Zhou, M.D., Ph.D., Department of Melanoma and Medical Oncology; Department of Immunology, University of Texas MD Anderson Cancer Center in Houston, TX. PUBLIC HEALTH RELEVANCE: MUC1 is a cell surface glycoprotein that is over-expressed and aberrantly glycosylated in many carcinoma. The tumor-associated antigens present on the cell surface are ideal targets for the development of effective immunotherapies, however, the exact structural features of the epitopes expressed by cancer cells are not known. It is our goal to develop the tools, a set of monoclonal antibodies, which can specifically identify these cell surface glycopeptide epitopes.