Surfaceomic technologies and antibodies to probe cell surface proteomes and their interactomes at unprecedented small scale and high-resolution

表面组学技术和抗体以前所未有的小规模和高分辨率探测细胞表面蛋白质组及其相互作用组

• 批准号: 10552328
• 负责人: JAMES A WELLS
• 金额: $ 57.22万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10552328
• 关键词: Advanced Development; Antibodies; Antineoplastic Agents; Bacteriophages; Biological Markers; Cell Surface Proteins; Cell membrane; Cell surface; Cells; Complex; Cytokine Receptors; Development; Disease; Drug Targeting; Energy Transfer; Environment; Epidermal Growth Factor Receptor; Family; Generations; Goals; Grant; Health; Immunology; Label; Malignant Neoplasms; Maps; Mediating; Membrane; Membrane Proteins; Methods; Minority; Modeling; Nerve Degeneration; Neurology; Oncogenes; Patients; Phenotype; Population; Process; Protein Secretion; Proteome; Proteomics; Recombinants; Regulation; Resolution; Sampling; Science; Signal Transduction; T-Lymphocyte; Techniques; Technology; Vision; Yeasts; cell type; improved; induced pluripotent stem cell; protein complex; receptor

Title: Surfaceomic technologies and antibodies to probe cell surface proteomes and their interactomes at unprecedented small scale and high-resolution Project summary The cell surfaceome is the primary hub that allows cells to sense and respond to changes in their environment, yet only a minority of the estimated 4000-5000 plasma membrane and secreted proteins have been functionally characterized. We understand even less about coordinate regulation of the surfaceome, and specifically how the composition, complexes, distribution, and function of membrane proteins are altered to collectively mediate changes to cellular phenotypes. Our long-range goal is to systematically understand how cells remodel their membrane proteome (surfaceome) in health and disease, and to develop antibodies that probe and modulate these processes. With the support of the R35 MIRA in the last five years, we have extensively characterized the surfaceome changes induced by oncogene transformation leading to discovery of new biomarkers, cancer drug targets, and generation of highly specific antibodies. In the next five years, we will develop new surfaceome technologies for small scale analysis of cell populations, particularly in neurodegeneration using iPSC derived models and patient-derived samples. We will define the cell surface protein complexes and interactomes using a high-resolution proximity labeling method enabled by Dexter Energy Transfer (DET). We plan to interrogate the interactome of EGFR family receptors in i ii iii cancer, and cytokine receptors in T-cell Ir activation. We believe the technologies for small scale surfaceomics and interactomics will dramatically improve our abilities to map cell surfaces and their intreactomes in diseases ranging from cancer, immunology, neurology, and more. We have produced recombinant Figure 1. Overview of development of surfaceomics technologies. antibodies to 100s of cell surface proteins using phage, yeast, and mammalian display, and we anticipate the expanded efforts in understanding the surfaceome will advance the development of highly selective antibodies that probe and modulate cellular states. We began this vision in our first R35 grant by defining the surfaceomes induced by specific oncogenes in cancer. Next, we will greatly expand surfaceomics science to much smaller scales of specialized and primary cells, and much higher resolution analysis of signaling interactomes.

表面组学技术和抗体探测细胞表面蛋白质组及其相互作用组 以前所未有的小规模和高分辨率 项目摘要 细胞表面组是细胞感知和响应环境变化的主要枢纽， 然而，在估计的4000-5000种质膜和分泌蛋白质中，只有少数在功能上是 表征了我们对表面基因组的协调调节了解得更少，特别是 膜蛋白的组成、复合物、分布和功能被改变， 细胞表型的变化。我们的长期目标是系统地了解细胞如何重塑 他们的膜蛋白质组（表面组）在健康和疾病，并开发抗体，探针和 调节这些过程。在过去五年中，在R35 MIRA的支持下，我们广泛地 表征了癌基因转化诱导的表面组变化，从而发现了新的 生物标志物、癌症药物靶标和高度特异性抗体的产生。未来五年，我们将 开发新的表面组技术，用于小规模细胞群分析，特别是 使用iPSC衍生的模型和患者衍生的样品来评估神经变性。我们将定义细胞表面 蛋白质复合物和相互作用体，使用Dexter Energy的高分辨率邻近标记方法 转移（DET）。我们计划审问 EGFR家族受体相互作用组 癌症和T细胞中的细胞因子受体 IR activation.我们相信， 小规模表面组学和相互作用组学将 极大地提高了我们绘制细胞的能力 疾病中的表面及其内切体 从癌症，免疫学，神经学， 和更多.我们已经产生了重组体图1。表面组学技术的发展概况。 100种细胞表面蛋白的抗体， 噬菌体，酵母和哺乳动物展示，我们预计在理解表面组的扩大努力 将促进开发高选择性抗体，探测和调节细胞状态。我们开始 这一愿景在我们的第一个R35基金中通过定义癌症中特定癌基因诱导的表面体来实现。接下来我们 将极大地扩展表面组学科学到更小规模的特化和原代细胞， 信号相互作用组的更高分辨率分析。