Mouse models for study of the NLRP1 and CARD8 inflammasomes

摘要 炎性小体是多蛋白复合物，作为细胞溶质传感器发挥作用。因此，它们响应于 内源性和外源性因素导致的细胞代谢和完整性的损害和改变 “与神经系统相关的刺激”相反，异常和慢性炎性小体激活可导致 自身炎性疾病炎性体复合物的组装导致半胱天冬酶的自激活， IL-1β和IL-18的成熟，以及通常的热凋亡性细胞死亡。虽然在下游有一个共同点， 在炎性小体激活后的事件中，反应的特异性依赖于种系编码的 模式识别受体由特定的神经元相关刺激触发。许多细胞传感器 如NLRP 3属于NLR基因家族，我们对个体贡献的理解 已经收集到炎性小体对先天免疫反应对过多环境挑战的影响 来自于对缺乏各种NLR的小鼠品系的研究。相比之下，关于 这两种炎性小体对免疫应答的贡献是本提案的重点：NLRP 1和 CARD8.缺乏这两种相关炎性小体信息的主要因素是可用性有限 动物模型，以评价人炎性小体的功能。职能的主要区别 人类和小鼠NLRP 1通过NLRP 1蛋白质结构和表达的物种差异而被提出 格局在CARD 8的情况下，已经对暂时指定给该蛋白质的功能进行了体内验证。 在小鼠/啮齿类动物中不可能，因为这些物种中不存在该基因。因此，对这种炎性小体的研究 几乎完全依赖于使用人细胞系和组织的体外和离体研究。我们建议 通过利用同线置换来产生表达以下蛋白的新小鼠品系， 人NLRP 1和CARD 8。

ABSTRACT Inflammasomes are multiprotein complexes that function as cytosolic sensors. As such, they respond to compromise and alteration of cellular metabolism and integrity resulting from both endogenous and exogenous “danger-associated stimuli”. Conversely, aberrant and chronic inflammasome activation can contribute to autoinflammatory diseases. Assembly of the inflammasome complex results in autoactivation of caspases, maturation of IL-1β and IL-18, and often pyroptotic cell death. While there is a downstream commonality in the events following inflammasome activation, the specificity of the response is dependent on germline-encoded pattern recognition receptors triggered by the specific danger-associated stimuli. Many of the cellular sensors such as NLRP3 belong to the NLR gene family, and much of our understanding of the contribution of individual inflammasomes to the innate immune responses to a plethora of environmental challenges has been gleaned from the study of mouse lines lacking various NLRs. In contrast, there is limited information regarding the contribution to immune responses of two inflammasomes that are the focus of this proposal: NLRP1 and CARD8. A major factor in the lack of information on these two related inflammasomes is the limited availability of animal models to evaluate the function of the human inflammasome. Major differences in the functions of human and mouse NLRP1 are suggested by species differences in NLRP1 protein structure and expression pattern. In the case of CARD8, in vivo validation of the functions tentatively assigned to this protein have been impossible in mice/rodents because the gene is absent in these species. Thus, the study of this inflammasome has depended almost entirely on in vitro and ex vivo studies using human cell lines and tissue. We propose to address this gap in our knowledge by utilizing syntenic replacement to generate novel mouse lines expressing human NLRP1 and CARD8.