Integrated Functional Genomics Analysis of Druggable T Cell-Activating Genes

所有主要基因组的测序完成，为基因组范围内的研究提供了新的视角。 序列和结构。然而，我们对如此大的基因组进行功能分析的能力仍然是有限的。 有限公司为了开发更有效的功能基因注释的新技术，我们建议， 一个多学科的方法，使用T细胞受体（TCR）信号作为一个例子， 生理学和病理学上重要的细胞过程， 但在可用数据能够进行综合大规模分析的情况下， 本案无关如果成功，我们的“系统”方法将广泛适用于许多领域， 生物学有望通过其被广泛采用来加速生物学知识的产生。 其他人，深刻影响大型科学和患者社区。作为 在适应性免疫系统中，T细胞是我们防御病原体、癌症的关键介质 细胞和不希望的移植排斥反应。T细胞功能受损可导致 免疫缺陷，促进感染或癌症。T细胞过度活跃或失调是一个主要的 过敏、自身免疫或神经退行性疾病的原因。这些严重的疾病 导致个人残疾、生活质量下降、无法自我维持、过早死亡 给社会和卫生保健系统带来沉重的经济负担。通过识别新的基因 介导T细胞活化并分析其分子功能，因此我们的研究将 促进我们对疾病中T细胞功能和功能障碍的理解， 一般来说，转导。通过关注功能可以被调节的药物基因， 通过小分子或生物疗法，我们的目标是加速开发新的， 针对严重免疫疾病或癌症的改进和更具成本效益的疗法， 大量的患者群体。设计和验证我们的创新，综合，多学科 基因发现和分析的方法，我们将首先确定新的TCR信号转导基因在一个新的研究领域。 在靶向5000个靶点阵列化shRNA文库的T细胞中进行高通量RNA干扰筛选 药物基因我们将使用慢病毒shRNA递送来克服以下挑战： 非粘附 生长和T细胞对常规机器人siRNA递送方法的相对抗性， 和新颖的屏幕设计和分析策略，以最大限度地提高灵敏度， 积极率。我们接下来将结合联合收割机基因表达谱，磷酸酪氨酸蛋白质组学， 单倍型关联作图，数量性状位点，全基因组关联作图 和基因表达QTL数据与基因/蛋白质网络和途径分析， 结合了联合收割机最高新奇和最大体内相关性的命中，用于详细随访。

The completed sequencing of all major genomes provided genome-wide insight into gene sequence and structure. However, our ability to functionally analyze such large gene sets is still limited. To develop novel technologies for more efficient functional gene annotation, we propose a multidisciplinary approach, using T cell receptor (TCR) signaling as an example of a physiologically and pathologically important cellular process that is still incompletely understood, but where available data enable integrated large-scale analysis to maximize novelty and in vivo relevance. If successful, the broad applicability of our "systems" approach to many areas of biology promises to accelerate generation of biological knowledge at large through its adoption by others, profoundly impacting large scientific and patient communities. As critical components of the adaptive immune system, T cells are key mediators of our defense against pathogens, cancer cells and, undesired, of transplant rejection. Impaired T cell function can result in immunodeficiency, promoting infections or cancer. T cell hyperactivity or deregulation is a main cause for allergies, autoimmune or neurodegenerative diseases. These serious disorders contribute to individual disability, loss of quality of life, inability to self-sustain, premature death and high economic burden for society and health care system. By identifying novel genes mediating T cell activation and analyzing their molecular functions, our proposed studies will thus promote our understanding of T cell function and malfunction in disease, and of signal transduction in general. By focusing on druggable genes whose functions can be modulated through small molecule or biological therapeutics, we aim to accelerate the development of new, improved and more cost-effective therapies for severe immune disorders or cancer that affect large patient populations. To devise and validate our innovative, integrated, multidisciplinary approach for gene discovery and analysis, we will first identify novel TCR signaling genes in a high-throughput RNA interference screen in T cells of an arrayed shRNA library that targets 5,000 druggable genes. We will use lentiviral shRNA delivery to overcome the challenges of nonadherent growth and relative T cell resistance to conventional robotic siRNA delivery methods, and novel screen designs and analysis strategies to maximize sensitivity and minimize false positive rates. We will next combine gene expression profiling, phosphotyrosine-proteomics, haplotype association mapping, quantitative trait locus (QTL), genome-wide associtation mapping and gene expression QTL data with gene/protein network and pathway analyses to prioritize those hits which combine highest novelty with maximal in vivo relevance for detailed follow-up.