Validation of a Genetic-based Biomarker Panel for Stratification of Mortality Risk in ARDS Patients

项目总结： 新冠肺炎疫情极大地突显了急性呼吸窘迫综合征未得到满足的严重需求，包括缺乏 FDA批准的有效药物干预措施，解决ARDS死亡率问题。ARDS表型 异质性，失调炎症的复杂性，以及缺乏可预测的生物标志物，这些都是 导致ARDS治疗临床试验失败。预测性生物标志物，无论是基于蛋白质还是基于基因组， 识别特定的ARDS亚型和可能对特定的ARDS疗法有反应的人，可能 显著影响评估新疗法的有效临床试验设计，从而使 急性呼吸窘迫综合征试验结果。我们小组长期以来一直支持使用基因组密集型方法来识别 多个新的ARDS治疗靶点。我们之前发现了eNAMPT(细胞外烟酰胺 磷酸核糖转移酶)作为ARDS治疗的新靶点/基因，作为损伤相关的 分子模式蛋白(DAMP)和Toll样受体4的配体(TLR4)。NAMPT SNPs预测ARDS 严重程度和eNAMPT放大失调的肺/全身炎症反应，导致多发性 器官损伤/衰竭。我们证明了人源化的eNAMPT中和单抗作为一种治疗方法的实用性。 在ARDS和其他炎症性情况下的策略。血浆eNAMPT以及IL-6、IL-8、IL-1RA、MIF和 Ang-2对28天的ARDS死亡率有很高的预测作用。我们还鉴定了选择素P配体基因的变异体 (SELPLG)编码P-选择素糖蛋白配体1(PSGL1)和P-选择素基因(SELP)与 黑人对ARDS的易感性增加。PSGL1/P-选择素相互作用是肺部炎症的关键途径 白细胞转运、血小板聚集和血栓形成。血浆PSGL1和P-选择素水平显著升高 脓毒症、急性呼吸窘迫综合征和新冠肺炎肺炎患者的升高和PSGL1抑制(单抗、TSGL-Ig)显著升高 减轻ARDS的临床前肺损伤。这项R-21应用程序将使用900多个血浆样本和 NHLBI急性肺损伤(PEAL)预防和早期治疗的基因分型结果 系统性早期神经肌肉阻断(ROSE)研究的网络重新评估(参见NHLBI BioLINCC Letter)。 在未来ARDS的设计中，我们将验证两种高度新颖的分层工具，以改进患者分层 针对eNAMPT/TLR4和PSGL1/P-选择素相互作用的临床试验。特定目标(SA)#1将制定一个 基于基因型的生物标记物分析结合：i)用血浆仔细选择SELPLG/SELP变异体 PSGL1/P-选择素水平，以及ii)与血浆eNAMPT水平相关的NAMPT变体。这些基因型会识别 ARDS受试者作为未来针对PSGL-1/P-选择素相互作用的临床试验的候选对象 ENAMPT/TLR4信号通路。SA#2将验证七个生物标志物小组的预测能力 (IL-6、IL-8、IL-1RA、PSGL-1、IL-1β、Ang-2)对急性呼吸窘迫综合征病死率的影响。成功地完成了这一高度 创新的R21赠款将产生一种可利用的新的“关注点”药物遗传浓缩工具 设计针对PSGL1/P-选择素和eNAMPT/TLR4相互作用的人类ARDS临床试验。

PROJECT SUMMARY: The COVID-19 pandemic has dramatically highlighted serious unmet needs of ARDS including the absence of effective FDA-approved pharmacologic interventions that address ARDS mortality. ARDS phenotype heterogeneity, the complexity of dysregulated inflammation, and the absence of predictive biomarkers have all contributed to failed ARDS therapeutic clinical trials. Predictive biomarkers, either protein- or genomic-based, that identify specific ARDS sub-phenotypes and likely responders to specific ARDS therapeutics, could significantly influence effective clinical trial designs to assess novel therapeutics and therefore benefit the outcomes of ARDS trials. Our group has long championed the utility of genomic-intensive approaches to identify multiple novel ARDS therapeutic targets. We previously identified eNAMPT (extracellular nicotinamide phosphoribosyl transferase) as a novel ARDS therapeutic target/gene which serves as a damage-associated molecular pattern protein (DAMP) and ligand for Toll-like receptor 4 (TLR4). NAMPT SNPs predict ARDS severity and eNAMPT amplifies dysregulated lung/systemic inflammatory responses that contribute to multi- organ injury/failure. We demonstrated the utility of a humanized eNAMPT-neutralizing mAb as a therapeutic strategy in ARDS and other inflammatory conditions. Plasma eNAMPT, along with IL-6, IL-8, IL-1RA, MIF, and Ang-2, was highly predictive of 28-day ARDS mortality. We also identified variants in selectin P ligand gene (SELPLG) encoding P-selectin glycoprotein ligand 1 (PSGL1), and P-selectin gene (SELP) as associated with increased susceptibility to ARDS in Blacks. PSGL1/P-selectin interactions are critical to lung inflammation via leukocyte trafficking, platelet aggregation, and thrombosis. Plasma PSGL1 and P-selectin levels are significantly elevated in sepsis, ARDS, and COVID-19 pneumonia patients and PSGL1 inhibition (mAb, TSGL-Ig) significantly attenuates preclinical lung injury in ARDS. This R-21 application will utilize over 900 plasma samples and genotyping results available from the NHLBI Prevention and Early Treatment of Acute Lung Injury (PETAL) Network Reevaluation of systemic Early neuromuscular blockade (ROSE) study (see NHLBI BioLINCC letter). We will validate two highly novel stratification tools to improve patient stratification in the design of future ARDS clinical trials targeting eNAMPT/TLR4 and PSGL1/P-Selectin interactions. Specific Aim (SA) #1 will develop a genotype-based biomarker assay combining: i) carefully selected SELPLG/SELP variants with plasma PSGL1/P-selectin levels, and ii) NAMPT variants with plasma eNAMPT levels. These genotypes will identify ARDS subjects as candidates for future clinical trials targeting PSGL-1/P-selectin interactions and the eNAMPT/TLR4 signaling pathway. SA #2 will validate the predictive capacity of a seven-biomarker panel (eNAMPT, IL-6, IL-8, IL-1RA, PSGL-1, IL-1β, Ang-2) for ARDS mortality. Successful completion of this highly innovative R21 grant will generate a novel ‘point of care’ pharmacogenetic enrichment tools to be leveraged in designing human ARDS clinical trials targeting PSGL1/P-selectin and eNAMPT/TLR4 interactions.