Mapping a clinically significant internalizing tumor epitope space.

绘制具有临床意义的内化肿瘤表位空间。

• 批准号: 7196540
• 负责人: BIN LIU
• 金额: $ 26.51万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-07 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7196540
• 关键词: Address; Affinity; Androgens; Animal Model; Antibodies; Antigen Targeting; Antigens; Bacteriophages; Binding; Biological Assay; Biological Markers; Carbohydrates; Cell surface; Cells; Chemicals; Chemistry; Complementary DNA; Development; Diagnostic; Directed Molecular Evolution; Environment; Epitopes; Fluorescence-Activated Cell Sorting; Freezing; Gene Expression; Generations; Genes; Genomics; Goals; Growth; Health; Human; Immunoglobulin Variable Region; Immunologic Surveillance; In Situ; Individual; Knowledge; Libraries; Life; Malignant neoplasm of prostate; Maps; Mass Spectrum Analysis; Methodology; Methods; Molecular; Molecular Conformation; Monoclonal Antibodies; NIH Program Announcements; Neoplasm Metastasis; Noise; Numbers; Organ; Output; Pan Genus; Pathogenesis; Phage Display; Physiology; Post-Translational Protein Processing; Primary Neoplasm; Proteins; Range; Relative (related person); Research; Research Personnel; Sampling; Signal Transduction; Slide; Sorting - Cell Movement; Specificity; Structure; Surface; Surface Antigens; System; Techniques; Technology; Therapeutic; Tissues; Tumor Antigens; Tumor Cell Line; Tumor Markers; Variant; Work; Yeasts; base; cDNA Library; cancer cell; cancer type; cell type; clinically relevant; clinically significant; combinatorial; design; drug development; improved; laser capture microdissection; mRNA Expression; neoplastic; neoplastic cell; novel; novel diagnostics; prognostic; programs; protein aminoacid sequence; receptor mediated endocytosis; research study; selective expression; tool; tumor

DESCRIPTION (provided by applicant): The goal of this proposal is to develop antibody library-based methods which promote efficient identification of clinically relevant tumor specific cell surface antigens, methods which are applicable to the identification of cell type-specific lineage markers in general. The abnormal physiology of tumor cells is reflected in part in the altered chemical and molecular composition of their cell surface. Progressive changes in surface molecule expression allow tumor cells to respond efficiently to external signals for growth and survival, to interact with host tissues, to achieve metastasis, and to avoid immune surveillance. The identification and targeting of tumor-specific cell surface antigens, however, is hampered by the complexity of the epitope space at the tumor cell surface. In addition to proteins, relevant antigens include carbohydrates and other post-translational modification products that cannot be predicted from studies of genomic copy number or mRNA expression levels. This proposal aims to develop combinatorial antibody library-based strategies that allow efficient identification of tumor specific cell surface antigens for diagnostic, prognostic and therapeutic applications. Specifically, we aim (1) To develop a high throughput subtractive selection strategy based on flow cytometric sorting to significantly improve selection efficiency and to obtain greater numbers of tumor- targeting phage antibodies. (2) To identify clinically relevant tumor antigens by selecting phage antibodies against prostate cancer cells in situ, within their proper stromal contexts. We propose to combine antibody library technology with laser capture microdissection (LCM) to identify antibodies against individual tumor cells on tissue slides. The resulting antibodies will have a very high likelihood of recognizing clinically relevant tumor antigens. (3) To establish the molecular identify of tumor cell surface antigens recognized by tumor-specific antibodies. We have developed a novel combinatorial yeast cDNA display library. Specific antigen-antibody pairs will be identified from this library following flow sorting. In parallel, we will pursue the analysis of tumor antigens by mass spectrometry methods which are capable of analyzing post-translational modifications. This study will increase our knowledge of tumor physiology and will facilitate the design of effective therapy; it will also help determine the exact contribution of post-translational modifications to the final makeup of the tumor epitope space.

描述（由申请人提供）：该提案的目的是开发基于抗体库的方法，这些方法促进有效鉴定临床相关的肿瘤特异性细胞表面抗原，这些方法通常适用于鉴定细胞类型特异性谱系标记。肿瘤细胞的异常生理反映在其细胞表面的化学和分子组成改变中。表面分子表达的进行性变化使肿瘤细胞能够对外部信号有效响应，以供生长和生存，与宿主组织相互作用，实现转移并避免免疫监测。然而，肿瘤特异性细胞表面抗原的鉴定和靶向受肿瘤细胞表面的表位空间的复杂性的阻碍。除蛋白质外，相关的抗原还包括碳水化合物和其他翻译后修饰产物，这些产品无法从基因组拷贝数或mRNA表达水平的研究中预测。该建议旨在开发基于组合库的组合抗体策略，以有效地识别特异性细胞表面抗原用于诊断，预后和治疗应用。具体而言，我们的目的是基于流式细胞仪排序制定高吞吐量减去选择策略，以显着提高选择效率并获得更多的肿瘤靶向噬菌体抗体。 （2）通过在适当的基质环境中选择针对前列腺癌细胞的噬菌体抗体来鉴定临床相关的肿瘤抗原。我们建议将抗体文库技术与激光捕获微分解（LCM）相结合，以鉴定针对组织载玻片上单个肿瘤细胞的抗体。所得的抗体将具有很高的识别临床相关肿瘤抗原的可能性。 （3）建立通过肿瘤特异性抗体识别的肿瘤细胞表面抗原的分子鉴定。我们已经开发了一种新型的组合酵母cDNA展示库。在流动后，将从该库中识别特定的抗原抗体对。同时，我们将通过质谱法对肿瘤抗原进行分析，该方法能够分析翻译后修饰。这项研究将提高我们对肿瘤生理学的了解，并促进有效治疗的设计；它还将有助于确定翻译后修饰对肿瘤表位空间最终构成的确切贡献。