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: 7367198
• 负责人: BIN LIU
• 金额: $ 34.84万
• 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/7367198
• 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

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 combinatiroal 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.

该提案的目标是开发基于抗体库的方法，以促进有效识别 临床相关肿瘤特异性细胞表面抗原的鉴定，适用于鉴定的方法 一般细胞类型特异性谱系标记。肿瘤细胞的生理异常部分体现在 细胞表面的化学和分子组成发生改变。表面的渐进变化 分子表达使肿瘤细胞能够有效地响应生长和生存的外部信号， 与宿主组织相互作用，实现转移，并避免免疫监视。身份识别和 然而，表位的复杂性阻碍了肿瘤特异性细胞表面抗原的靶向 肿瘤细胞表面的空间。除蛋白质外，相关抗原还包括碳水化合物和其他 无法通过基因组拷贝数研究预测的翻译后修饰产物或 mRNA 表达水平。该提案旨在开发基于组合抗体库的策略 能够有效识别肿瘤特异性细胞表面抗原，用于诊断、预后和治疗 应用程序。具体来说，我们的目标是（1）开发一种基于 流式细胞术分选显着提高选择效率并获得更多的肿瘤 靶向噬菌体抗体。 (2) 通过选择噬菌体抗体来鉴定临床相关的肿瘤抗原 在适当的基质环境中原位对抗前列腺癌细胞。我们建议结合抗体 结合激光捕获显微切割 (LCM) 的库技术可识别针对个体肿瘤的抗体 组织玻片上的细胞。由此产生的抗体将有很高的可能性被临床识别 相关肿瘤抗原。 (3) 建立肿瘤细胞表面抗原识别的分子鉴定 肿瘤特异性抗体。我们开发了一种新型组合酵母 cDNA 展示文库。具体的 经过流式分选后，将从该文库中鉴定出抗原-抗体对。与此同时，我们将追求 通过质谱方法分析肿瘤抗原，该方法能够分析翻译后 修改。这项研究将增加我们对肿瘤生理学的了解，并将有助于设计 有效的治疗；它还将有助于确定翻译后修饰对 肿瘤表位空间的最终组成。