Department of Anesthesiology and Critical Care Medicine, The Children's Hospital of Philadelphia and University of Pennsylvania, Philadelphia, Pennsylvania.
Proteomics Core, The Children's Hospital of Philadelphia, Philadelphia, Pennsylvania.
Am J Physiol Lung Cell Mol Physiol. 2022 Mar 1;322(3):L365-L372. doi: 10.1152/ajplung.00164.2021. Epub 2022 Jan 5.
Both sepsis and acute respiratory distress syndrome (ARDS) rely on imprecise clinical definitions leading to heterogeneity, which has contributed to negative trials. Because circulating protein/DNA complexes have been implicated in sepsis and ARDS, we aimed to develop a proteomic signature of DNA-bound proteins to discriminate between children with sepsis with and without ARDS. We performed a prospective case-control study in 12 children with sepsis with ARDS matched to 12 children with sepsis without ARDS on age, severity of illness score, and source of infection. We performed co-immunoprecipitation and downstream proteomics in plasma collected ≤ 24 h of intensive care unit admission. Expression profiles were generated, and a random forest classifier was used on differentially expressed proteins to develop a signature which discriminated ARDS. The classifier was tested in six independent blinded samples. Neutrophil and nucleosome proteins were over-represented in ARDS, including two S100A proteins, superoxide dismutase (SOD), and three histones. Random forest produced a 10-protein signature that accurately discriminated between children with sepsis with and without ARDS. This classifier perfectly assigned six independent blinded samples as having ARDS or not. We validated higher expression of the most informative discriminating protein, galectin-3-binding protein, in children with ARDS. Our methodology has applicability to isolation of DNA-bound proteins from plasma. Our results support the premise of a molecular definition of ARDS, and give preliminary insight into why some children with sepsis, but not others, develop ARDS.
脓毒症和急性呼吸窘迫综合征 (ARDS) 均依赖于不精确的临床定义,导致异质性,这导致了负面试验。由于循环蛋白/DNA 复合物与脓毒症和 ARDS 有关,我们旨在开发一种 DNA 结合蛋白的蛋白质组学特征,以区分伴有和不伴有 ARDS 的脓毒症儿童。我们对 12 例伴有 ARDS 的脓毒症儿童和 12 例不伴有 ARDS 的脓毒症儿童进行了前瞻性病例对照研究,这些儿童的年龄、疾病严重程度评分和感染源相匹配。我们在入住重症监护病房≤24 小时内采集血浆进行共免疫沉淀和下游蛋白质组学分析。生成表达谱,并使用差异表达蛋白的随机森林分类器开发区分 ARDS 的特征。该分类器在六个独立的盲样中进行了测试。中性粒细胞和核小体蛋白在 ARDS 中过度表达,包括两种 S100A 蛋白、超氧化物歧化酶 (SOD) 和三种组蛋白。随机森林产生了一个由 10 种蛋白组成的特征,可准确区分伴有和不伴有 ARDS 的脓毒症儿童。该分类器完美地将六个独立的盲样分配为患有或不患有 ARDS。我们验证了 ARDS 儿童中 galectin-3 结合蛋白的表达更高。我们的方法适用于从血浆中分离 DNA 结合蛋白。我们的结果支持 ARDS 的分子定义的前提,并初步探讨了为什么一些脓毒症儿童但不是其他儿童会发展为 ARDS。