Key Points NLRP3 R259W homozygous pigs can be generated by somatic cell nuclear transfer. NLRP3 R259W homozygous pigs can survive to adulthood and give birth to offspring. Both homozygous- and heterozygous-mutated pigs show systemic spontaneous inflammation. The NLRP3 inflammasome is associated with a variety of human diseases, including cryopyrin-associated periodic syndrome (CAPS). CAPS is a dominantly inherited disease with NLRP3 missense mutations. Currently, most studies on the NLRP3-inflammasome have been performed with mice, but the activation patterns and the signaling pathways of the mouse NLRP3 inflammasome are not always identical with those in humans. The NLRP3 inflammasome activation in pigs is similar to that in humans. Therefore, pigs with precise NLRP3-point mutations may model human CAPS more accurately. In this study, an NLRP3 gain-of-function pig model carrying a homozygous R259W mutation was generated by combining CRISPR/Cpf1-mediated somatic cell genome editing with nuclear transfer. The newborn NLRP3 R259W homozygous piglets showed early mortality, poor growth, and spontaneous systemic inflammation symptoms, including skin lesion, joint inflammation, severe contracture, and inflammation-mediated multiorgan failure. Severe myocardial fibrosis was also observed. The tissues of inflamed skins and several organs showed significantly increased expressions of NLRP3, Caspase-1, and inflammation-associated cytokines and factors (i.e., IL-1β, TNF-α, IL-6, and IL-17). Notably, approximately half of the homozygous piglets grew up to adulthood and even gave birth to offspring. Although the F1 heterozygous piglets showed improved survival rate and normal weight gain, 39.1% (nine out of 23) of the piglets died early and exhibited spontaneous systemic inflammation symptoms. In addition, similar to homozygotes, adult heterozygotes showed increased delayed hypersensitivity response. Thus, the NLRP3 R259W pigs are similar to human CAPS and can serve as an ideal animal model to bridge the gap between rodents and humans.

中文翻译：

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携带功能获得性 NLRP3 纯合突变的工程猪可以存活到成年并准确再现人类全身性自发性炎症反应

关键点 NLRP3 R259W 纯合猪可以通过体细胞核移植产生。NLRP3 R259W纯合猪可以存活到成年并生育后代。纯合子和杂合子突变猪都表现出全身性自发性炎症。NLRP3 炎症小体与多种人类疾病有关，包括冷热蛋白相关周期性综合征 (CAPS)。CAPS 是一种具有 NLRP3 错义突变的显性遗传疾病。目前，大多数关于 NLRP3 炎症小体的研究都是在小鼠身上进行的，但小鼠 NLRP3 炎症小体的激活模式和信号通路并不总是与人类相同。猪的 NLRP3 炎症小体激活与人类相似。因此，具有精确 NLRP3 点突变的猪可以更准确地模拟人类 CAPS。在这项研究中，通过将 CRISPR/Cpf1 介导的体细胞基因组编辑与核转移相结合，产生了一个携带纯合 R259W 突变的 NLRP3 功能获得猪模型。新生 NLRP3 R259W 纯合仔猪表现出早期死亡率、生长不良和自发性全身炎症症状，包括皮肤病变、关节炎症、严重挛缩和炎症介导的多器官衰竭。还观察到严重的心肌纤维化。发炎皮肤和几个器官的组织显示出 NLRP3、Caspase-1 和炎症相关细胞因子和因子（即 IL-1β、TNF-α、IL-6 和 IL-17）的表达显着增加。值得注意的是，大约一半的纯合仔猪长大到成年，甚至生下了后代。尽管 F1 杂合仔猪的存活率提高，体重增加正常，39.1%（23 头中的 9 头）仔猪过早死亡并表现出自发性全身炎症症状。此外，与纯合子类似，成年杂合子显示出增加的延迟超敏反应。因此，NLRP3 R259W 猪与人类 CAPS 相似，可以作为理想的动物模型来弥合啮齿动物和人类之间的差距。