Mouse models for study of the NLRP1 and CARD8 inflammasomes

摘要 炎性小体是一种多蛋白复合体，起到胞液感受器的作用。因此，他们会对 内源性和外源性导致的细胞代谢和完整性的折衷和改变 “危险相关的刺激”。相反，异常和慢性炎症性小体激活可导致 自身炎症性疾病。炎症体复合体的组装导致半胱氨酸天冬氨酸酶的自动激活， IL-1β和IL-18成熟，常有嗜热性细胞死亡。虽然在下游有一个共同点，但 炎性小体激活后的事件，反应的特异性取决于生殖线编码 由特定危险相关刺激触发的模式识别受体。许多细胞传感器 如NLRP3属于NLR基因家族，我们对个体的贡献了解很多 已收集到针对过多环境挑战的先天免疫反应的炎症体 来自对缺乏各种NLR的小鼠品系的研究。相比之下，关于 这两个炎症体对免疫反应的贡献是这一提议的重点：NLRP1和NLRP1 CARD8。缺乏关于这两个相关炎症体的信息的一个主要因素是可获得性有限 以评估人类炎症小体的功能。在功能上的主要差异 人类和小鼠NLRP1的蛋白质结构和表达存在物种差异 图案。在CARD8的情况下，已经对初步分配给该蛋白质的功能进行了体内验证 在老鼠/啮齿动物中是不可能的，因为这种基因在这些物种中缺失。因此，对这个炎症体的研究 几乎完全依赖于使用人类细胞系和组织进行的体外和体外研究。我们建议 通过利用共线替换来产生新的鼠线表达来填补我们知识中的这一缺口 人类NLRP1和CARD8。

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.