Signaling Core

这一提议代表了一种高度创新的、或许是独一无二的方法，为 了解人类柯萨奇病毒19驱动的病理学的发病机制。在这里，我们带来了三个不同的科学 以一种新颖和协调的方式共同分析先天免疫反应的关键步骤 新冠肺炎感染。NLRP3炎症体通路的基础研究既深入又广泛 鉴于Ulevitch实验室在研究NLR家族各种成员方面近20年的经验 通径分析水平。同时利用小鼠细胞研究的信息(遗传、生化和 细胞生物学)这里的焦点是人类M？在那里，Ulevitch实验室已经建立了广泛的方法 免疫学、细胞生物学和遗传学方法。诺兰实验室率先使用高含量 用于分析细胞和组织的成像方法。高参数技术最大限度地扩大了 分析样本中的免疫特征，以便快速发现。CyTOF为数百万个小区提供42个通道的信息 血。Codex用&gt；60参数空间解析组织中的单个细胞，而ViralMIBI带来病毒 从核酸检测到以抗体为基础的多参数成像，确定了感染的细胞定位。 将这些技术与SARS-CoV-2挑战模型和协作交叉相结合也将产生 在鉴定与疾病表型和分子最接近的小鼠模型中 新冠肺炎病在人类身上的特征。如果被认为是有用的，我们也可以使用信息和/或老鼠 来自Beutler实验室的正向遗传努力的菌株。后者一直是 斯克里普斯U19计划自2001/2002年首次资助以来。

This proposal represents a highly innovative and perhaps unique approach to lay the foundation for understanding the pathogenesis of COVID19-driven pathologies in man. Here we bring three distinct scientific approaches together in a novel and coordinated fashion to analyze key steps in innate immune response to COVID-19 infection. The fundamental studies of the NLRP3 inflammasome pathway is both deep and broad given the nearly 20-year experience of the Ulevitch lab in studying various members of the NLR family at the level of pathway analysis. While drawing on information from studies in murine cells (genetic, biochemical and cell biologic) the focus here is on human M? where the Ulevitch lab has built a broad approach with immunologic, cell biologic and genetic approaches. The Nolan lab has pioneered the use of high content imaging approaches for analyzing cells and tissues. The high parameter technologies maximize the breadth of immune features analyzed in samples for rapid discovery. CyTOF gives 42-channel info for millions of cells in blood. CODEX spatially resolves single cells in tissues with >60 parameters, while ViralMIBI brings viral nucleic acid detection to antibody-based multiparameter imaging defining cellular niches of infection. Combining these technologies with SARS-CoV-2 challenge models and the Collaborative Cross will also result in the identification of mouse models that most closely match the disease phenotypes and molecular signatures of COVID-19 disease in humans. If deemed useful we can also use information and/or murine strains derived from forward genetic efforts in the Beutler lab. The latter has been an integral part of the Scripps U19 program since the initial funding in 2001/2002.