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.

标题：探测细胞表面蛋白质组及其相互作用的抗体技术和抗体 在前所未有的小规模和高分辨率 项目摘要 细胞表面组是一个主要的枢纽，允许细胞感知并响应其环境变化， 然而，估计的4000-5000个质膜和分泌蛋白的估计中只有少数是功能 特征。我们对Surfaceome的坐标调节的了解甚至更少，特别是如何 膜蛋白的成分，复合物，分布和功能被改变以集体介导 细胞表型的变化。我们的远程目标是系统地了解细胞如何重塑 它们在健康和疾病中的膜蛋白质组（表面体），并开发探测和 调节这些过程。在过去五年中R35 Mira的支持下，我们有广泛的 表征了通过癌基因转化引起的表面变化，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。在接下来的五年中，我们将 开发新的Surfaceome技术，用于细胞群体的小规模分析，尤其是在 使用IPSC得出的模型和患者衍生的样品的神经变性。我们将定义细胞表面 蛋白质复合物和相互作用使用由Dexter Energy启用的高分辨率接近标记方法 转移（det）。我们计划审问 I III III中EGFR家族受体的相互作用 T细胞中的癌症和细胞因子受体 ir 激活。我们相信 小规模的表面组学和相互作用组学 急剧提高了我们映射细胞的能力 疾病中的表面及其脑反应 癌症，免疫学，神经病学不等 还有更多。我们已经产生了重组图1。表面学技术的发展概述。 使用使用100秒细胞表面蛋白的抗体 噬菌体，酵母和哺乳动物展示，我们预计了解Surfaceome的努力不断扩大 将推进探测和调节细胞状态的高度选择性抗体的发展。我们开始了 在我们的第一个R35授予中，这种视野是通过定义癌症特异性癌基因引起的表面粒。接下来，我们 将大大扩展表面科学的专业和原代细胞的尺度大得多，而且很多 信号相互作用的高分辨率分析。