Paneth cells-derived IL-17A and liver ischemia reperfusion injury

肝脏缺血再灌注损伤是肝脏缺血再灌注损伤过程中和术后的重要临床问题 肝移植、肝切除和门静脉重建手术。不幸的是，没有治疗方法 预防或治疗肝脏IR损伤。此外，越来越清楚的是，肝脏IR损伤经常发生在肝脏， 导致显著的肝外远端器官损伤，包括肾、肠和肺损伤以及 全身炎症反应综合征和脓毒症。特别是，急性肾损伤的发生率， 肝切除率极高，在主要肝切除术或肝移植术后接近~50-80%。 我们以前的研究表明小肠隐窝潘氏细胞是肝外远端器官的起始细胞 小鼠肝脏IR后的损伤。此外，小肠潘氏细胞是促胰岛素分泌的主要来源。 在小鼠中产生肝和肝外损伤所需的炎性细胞因子IL-17 A。但对于 这些研究的临床转化导致治疗，这是至关重要的，我们确定是否潘氏细胞 在人类肝脏IR后，IL-17 A的失调和脱颗粒以及潘氏细胞释放也发生 和非人灵长类动物肝IR损伤模型。在这个建议中，我们将阐明小肠是否 人和非人灵长类动物肝脏IR后潘氏细胞失调并释放促炎性IL-17 A 损伤以及IL-17 A中和是否减轻肝损伤以及肝外器官损伤 令人兴奋的初步数据表明，食蟹猴肝脏IR引起深刻的 潘氏细胞脱粒和潘氏细胞IL-17 A诱导与快速肾脏和小肠损伤偶联。 此外，我们的初步数据表明，用IL-17 A中和抗体处理的食蟹猴 肝脏IR损伤明显减少，术后阿基减少。初步数据还表明， 人小肠缺血损伤后潘氏细胞凋亡。根据这些初步调查结果，我们 假设肝IR损伤导致肠Paneth细胞失调/脱颗粒和Paneth细胞- 衍生的IL-17 A诱导导致肠道炎症和细胞凋亡，随后肝细胞凋亡加重， 损伤和诱发急性肾损伤。我们还假设IL-17 A中和作用会减弱 非人灵长类动物肝脏IR后的肝、肠和肾损伤。我们将在体内（高度 翻译食蟹猴肝IR和人肝移植研究）和体外（新鲜分离 食蟹猴和人潘氏细胞）模型，以进一步阐明机制和潜力 通过测试以下三个具体目标，对肝IR后肠和肾损伤的治疗进行了研究。 目的#1：证明肝脏IR损伤后的远端器官损伤和潘氏细胞脱粒。 目的#2：证明肝脏IR损伤后潘氏细胞介导的IL-17 A释放。 目的#3：开发减少非人灵长类动物肝脏IR损伤诱导的肝和肝外损伤的疗法 器官损伤

Liver ischemia and reperfusion (IR) injury is a clinically significant problem during and after liver transplantation, hepatic resection and portal vein reconstruction surgery. Unfortunately, there is no therapy to prevent or treat hepatic IR injury. Furthermore, it is becoming increasingly clear that liver IR injury frequently results in significant extra-hepatic remote organ injury including kidney, intestinal and lung injury as well as a systemic inflammatory response syndrome and sepsis. In particular, the incidence of acute kidney injury after liver resection is extremely high and approaches ~50-80% after major hepatic resection or liver transplantation. Our previous studies implicate small intestinal crypt Paneth cells as the initiator of extra-hepatic remote organ injury after liver IR in mice. Furthermore, small intestinal Paneth cells are the predominant source of pro- inflammatory cytokine IL-17A required for generating hepatic and extra-hepatic injury in mice. However, for clinical translation of these studies leading to therapy, it is critical that we determine whether Paneth cell dysregulation and degranulation as well as Paneth cell release of IL-17A also occur after hepatic IR in human and nonhuman primate liver IR injury models. In this proposal, we will elucidate whether small intestinal Paneth cells dysregulate and release pro-inflammatory IL-17A after human and nonhuman primate liver IR injury and whether IL-17A neutralization attenuates liver injury as well as extra-hepatic organ injury after nonhuman primate IR. Exciting preliminary data suggest that cynomolgus monkey liver IR causes profound Paneth cell degranulation and Paneth cell IL-17A induction coupled with rapid kidney and small intestine injury. Furthermore, our preliminary data suggest that cynomolgus monkeys treated with IL-17A neutralizing antibody had markedly less hepatic IR injury as well as reduced post-operative AKI. Preliminary data also suggest that human small intestine Paneth cells degranulate after ischemic injury. Based on these preliminary findings, we hypothesize that hepatic IR injury leads to intestinal Paneth cell dysregulation/degranulation and Paneth cell- derived IL-17A induction leading to intestinal inflammation and apoptosis, subsequent exacerbation of hepatic injury and induction of acute kidney injury. We also hypothesize that IL-17A neutralization will attenuate hepatic, intestinal and kidney injury after liver IR in nonhuman primates. We will utilize both in vivo (highly translational cynomolgus monkey liver IR and human liver transplant studies) and in vitro (freshly isolated cynomolgus monkey and human Paneth cells) models to further elucidate the mechanisms and potential therapy for intestinal and renal injury after hepatic IR by testing the following three specific aims. Aim #1: To demonstrate remote organ injury and Paneth cell degranulation after liver IR injury. Aim #2: To demonstrate Paneth cell-mediated IL-17A release after liver IR injury. Aim #3: To develop therapies to reduce non-human primate liver IR injury induced hepatic and extra-hepatic organ injury.