Department of Pathology, Boston Children's Hospital and Harvard Medical School, Boston, Massachusetts, USA,
Department of Pediatrics, Precision Vaccines Program, Boston Children's Hospital and Harvard Medical School, Boston, Massachusetts, USA.
Neonatology. 2022;119(2):193-203. doi: 10.1159/000520680. Epub 2022 Jan 24.
Current techniques to diagnose and/or monitor critically ill neonates with bronchopulmonary dysplasia (BPD) require invasive sampling of body fluids, which is suboptimal in these frail neonates. We tested our hypothesis that it is feasible to use noninvasively collected urine samples for proteomics from extremely low gestational age newborns (ELGANs) at risk for BPD to confirm previously identified proteins and biomarkers associated with BPD.
We developed a robust high-throughput urine proteomics methodology that requires only 50 μL of urine. We utilized the methodology with a proof-of-concept study validating proteins previously identified in invasively collected sample types such as blood and/or tracheal aspirates on urine collected within 72 h of birth from ELGANs (gestational age [26 ± 1.2] weeks) who were admitted to a single Neonatal Intensive Care Unit (NICU), half of whom eventually developed BPD (n = 21), while the other half served as controls (n = 21).
Our high-throughput urine proteomics approach clearly identified several BPD-associated changes in the urine proteome recapitulating expected blood proteome changes, and several urinary proteins predicted BPD risk. Interestingly, 16 of the identified urinary proteins are known targets of drugs approved by the Food and Drug Administration.
In addition to validating numerous proteins, previously found in invasively collected blood, tracheal aspirate, and bronchoalveolar lavage, that have been implicated in BPD pathophysiology, urine proteomics also suggested novel potential therapeutic targets. Ease of access to urine could allow for sequential proteomic evaluations for longitudinal monitoring of disease progression and impact of therapeutic intervention in future studies.
目前诊断和/或监测患有支气管肺发育不良(BPD)的危重新生儿的技术需要对体液进行有创采样,而这在这些脆弱的新生儿中并不理想。我们验证了一个假设,即使用从有发生 BPD 风险的极低出生体重儿(ELGAN)收集的非侵入性尿液样本进行蛋白质组学分析是可行的,以确认与 BPD 相关的先前鉴定的蛋白质和生物标志物。
我们开发了一种稳健的高通量尿液蛋白质组学方法,仅需要 50μL 的尿液。我们使用该方法进行了概念验证研究,验证了先前在侵入性采集的样本类型(如血液和/或气管吸出物)中鉴定的蛋白质,这些样本是在出生后 72 小时内从被收入单个新生儿重症监护病房(NICU)的 ELGAN (胎龄[26±1.2]周)收集的,其中一半最终发生了 BPD(n=21),而另一半作为对照(n=21)。
我们的高通量尿液蛋白质组学方法清楚地识别了尿液蛋白质组中与 BPD 相关的几种变化,这些变化与预期的血液蛋白质组变化相一致,并且几种尿蛋白可预测 BPD 风险。有趣的是,鉴定出的 16 种尿液蛋白是食品和药物管理局批准的药物的已知靶标。
除了验证先前在侵入性采集的血液、气管吸出物和支气管肺泡灌洗液中发现的与 BPD 病理生理学有关的大量蛋白质外,尿液蛋白质组学还提示了新的潜在治疗靶标。易于获取尿液可以允许进行连续的蛋白质组学评估,以便在未来的研究中对疾病进展和治疗干预的影响进行纵向监测。