Therapeutic efficacies of neutrophil elastase and RIP1 inhibitors in acute lung injury

中性粒细胞弹性蛋白酶和RIP1抑制剂治疗急性肺损伤的疗效

基本信息

批准号：
10311120
负责人：
HASEM HABELHAH
金额：
$ 23.18万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-12-02 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10311120
关键词：
Ablation Acute Lung Injury Adverse effects Affect Alveolar American Animal Model Apoptosis Attenuated Bacteria Biochemical Breathing CASP8 gene Cell Death Cell Survival Cessation of life Clinical Research Clinical Trials Consensus Development Effectiveness Endothelium Enzymes Event FDA approved Genetic Goals Human Human body Hypoxemia Incidence Inflammation Injury Invaded Japanese Knock-out Knockout Mice Lethal Dose 50 Leukocyte Elastase Leukocytes Lipopolysaccharides Lung Measures Mediating Metalloproteases Modeling Molecular Multicenter Studies Mus Neutrophil Activation Pathway interactions Peptide Hydrolases Perinatal Pharmacology Phase Phase III Clinical Trials Phosphotransferases Play Proteins Pulmonary Edema Pulmonary Inflammation RIPK1 gene RIPK3 gene Reactive Oxygen Species Receptor Inhibition Respiratory Failure Role Stains Study models Syndrome TNF gene Tissues Treatment Efficacy Treatment Protocols Ubiquitination United States Ventilator experimental study improved in vivo inflammatory milieu inhibitor insight interstitial kinase inhibitor lung injury mortality mouse model neutrophil neutrophil elastase inhibitor novel novel therapeutic intervention pathogen prevent public health relevance pulmonary function small molecular inhibitor ventilation

项目摘要

5. Project Summary/Abstract Acute lung injury (ALI) is a syndrome of respiratory failure characterized by protein-rich pulmonary edema that causes severe hypoxemia and labored breathing. In the United States, the annual incidence of ALI is about 200,000 with a mortality rate of 30-40%; yet, there is no FDA-approved pharmacological therapy for ALI. Neutrophils are the most abundant circulating white blood cells in the human body and provide the first line of defense against invading pathogens. In the lung, while neutrophils are essential for pathogen elimination, exuberant accumulation and prolonged survival of neutrophils in the interstitial and alveolar spaces contribute to ALI by releasing a variety of injurious molecules, including neutrophil elastase (NE), metalloproteases, reactive oxygen species (ROS), and other proteolytic enzymes. Although all these injurious molecules contribute to ALI, an increasing body of studies have demonstrated that NE is the most potent hydrolytic enzyme that plays a key role in lung alveolar injury, and that inhibition of NE with small molecular inhibitors [e.g. Sivelestat (Siv)] reduces measures of lung injury and inflammation in animal models of ALI. However, clinical studies have not yet provided a clear consensus. Whereas in Japanese phase III clinical trials, Siv improved pulmonary functions, in North American multicenter studies, Siv rather increased long-term mortality by unknown mechanisms. Thus, identification of molecular events associated with adverse effects of Siv in ALI is an important task for the development of novel and combination treatment regimens. Recently, in unbiased functional protease purification experiments, we discovered that NE cleaves receptor interacting protein 1 (RIP1), and that inhibition of NE with Siv leads to RIP1-dependent necroptosis in mouse and human neutrophils following treatment with lipopolysaccharide (LPS) or TNFα. Such RIP1-induced necroptosis can be inhibited by RIP1 kinase inhibitors (e.g. Nec-1s) or knockout (KO) of RIP3. We found that LPS/Siv-induced necroptosis was significantly inhibited in RIP3 KO neutrophils. Importantly, in a mouse model of ALI induced by 50% lethal dose of LPS, Siv in combination with Nec-1s resulted in 90% survival of the mice (P=0.001), whereas neither Siv nor Nec-1s alone afforded significant protection against the death. Thus, we hypothesize that inhibition of NE with Siv in ALI leads to RIP1-mediated necroptosis in neutrophils, which causes secondary inflammation and lung injury, and that inhibition of RIP1-mediated necroptosis significantly improves the effectiveness of NE inhibitors in ALI. Aim-1. Determine the therapeutic efficacies of RIP1 and NE inhibitors in lung injury and inflammation using mouse models of ALI. Aim-2. Determine how genetic ablation of NE or RIP3 affects lung injury and inflammation in ALI mice.

5。项目摘要/摘要急性肺损伤（ALI）是一种以富含蛋白质的肺水肿为特征的呼吸衰竭综合征引起严重的低氧血症并呼吸困难。在美国，阿里的年度事件大约 200,000，死亡率为30-40％；但是，ALI没有FDA批准的药物疗法。中性粒细胞是人体中最丰富的循环白细胞，并提供了第一行防御入侵病原体。在肺中，虽然中性粒细胞对于病原体的进化至关重要，但中性粒细胞在间质和牙槽空间中的繁殖积累和长期存活贡献通过释放各种意外分子，包括中性粒细胞弹性酶（NE），金属蛋白酶活性氧（ROS）和其他蛋白水解酶。尽管所有这些有害分子有助于ALI，越来越多的研究表明，NE是最有效的水解在肺肺泡损伤中起关键作用的酶，并用小分子抑制剂抑制NE [例如。 Sivelestat（SIV）]减少了ALI动物模型中肺损伤和炎症的测量。然而，临床研究尚未提供明确的共识。而在日本III期临床试验中，SIV 在北美多中心研究中，改善了肺功能，SIV相当提高了长期死亡率通过未知的机制。这是鉴定与SIV在ALI中的不良反应相关的分子事件是开发新颖和组合治疗方案的重要任务。最近，无偏见功能性蛋白酶纯化实验，我们发现NE裂解受体相互作用蛋白1 （RIP1），并且对NE抑制SIV会导致小鼠和人的RIP1依赖性坏死性用脂多糖（LPS）或TNFα治疗后的中性粒细胞。这种RIP1引起的坏死性可能是 RIP1激酶抑制剂（例如NEC-1S）或RIP3的基因敲除（KO）抑制。我们发现LPS/SIV诱导的在RIP3 KO中性粒细胞中，坏死性受到显着抑制。重要的是，在由ALI的小鼠模型中 50％致死剂量的LPS，SIV与NEC-1S结合，导致小鼠的生存率为90％（p = 0.001），而仅凭SIV和NEC-1S都没有为死亡提供重大保护。那我们假设在ALI中抑制NE的NE导致RIP1介导的中性粒细胞的坏死性，这会导致继发性炎症和肺损伤，以及RIP1介导的坏死的抑制显着改善了 NE抑制剂在ALI中的有效性。 AIM-1。确定RIP1和NE抑制剂在使用ALI小鼠模型的肺损伤和炎症。 AIM-2。确定NE或RIP3的遗传消融方式影响ALI小鼠的肺损伤和炎症。