Mouse models for study of the NLRP1 and CARD8 inflammasomes

用于研究 NLRP1 和 CARD8 炎性体的小鼠模型

• 批准号: 10493370
• 负责人: Beverly H Koller
• 金额: $ 19.44万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-22 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10493370
• 关键词: 129 Mouse; Address; Animal Model; Autoimmune Diseases; C-terminal; Caspase; Cell Death; Cells; Chronic; Complex; Development; Disease; Event; Future; Gene Family; Generations; Genes; Glean; Goals; Host Defense; Human; Human Cell Line; Human Genome; Immune; Immune response; In Vitro; Individual; Inflammasome; Inflammatory; Innate Immune Response; Interleukin-1 beta; Interleukin-18; Knowledge; Leucine-Rich Repeat; Mediating; Metabolism; Modeling; Molecular; Multiprotein Complexes; Mus; N-terminal; Orthologous Gene; Pattern; Pattern recognition receptor; Population; Positioning Attribute; Process; Protein Family; Proteins; Reporting; Rodent; Role; Specificity; Stimulus; Structure; Study models; Tissues; Validation; Work; autoinflammatory; body system; genetic manipulation; in vivo; innate immune pathways; marenostrin; microbial; mouse genome; mouse model; multicatalytic endopeptidase complex; novel; polypeptide; protein expression; protein structure; response; sensor; species difference; tool

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.

抽象的 炎症小体是多蛋白复合物，起到胞质传感器的作用。因此，他们回应 内源性和外源性导致的细胞代谢和完整性的损害和改变 “与危险相关的刺激”。相反，异常和慢性炎症小体激活可导致 自身炎症性疾病。炎性体复合物的组装导致半胱天冬酶的自动激活， IL-1β 和 IL-18 成熟，并且经常导致细胞焦亡。虽然下游存在共性 炎症体激活后发生的事件，反应的特异性取决于种系编码 由特定危险相关刺激触发的模式识别受体。许多蜂窝传感器 例如 NLRP3 属于 NLR 基因家族，我们对个体的贡献有很多了解 已收集到对多种环境挑战的先天免疫反应的炎症小体 来自对缺乏各种 NLR 的小鼠品系的研究。相比之下，关于 本提案重点关注的两种炎症小体对免疫反应的贡献：NLRP1 和 卡8。缺乏这两种相关炎症小体信息的一个主要因素是可用性有限 动物模型来评估人类炎症小体的功能。功能上的主要区别 NLRP1 蛋白结构和表达的物种差异表明人类和小鼠 NLRP1 存在差异 图案。就 CARD8 而言，暂定分配给该蛋白质的功能的体内验证已经过 在小鼠/啮齿动物中这是不可能的，因为这些物种中不存在该基因。因此，对这种炎症小体的研究 几乎完全依赖于使用人类细胞系和组织的体外和离体研究。我们建议 通过利用同线性替换来生成表达的新小鼠品系，弥补了我们知识上的这一空白 人类 NLRP1 和 CARD8。