Dried blood spot proteomics analysis of newborn screening cards to identify prognostic markers of SIDS risk

对新生儿筛查卡进行干血点蛋白质组学分析，以确定 SIDS 风险的预后标志物

基本信息

批准号：
10734386
负责人：
ROBIN Lynn HAYNES
金额：
$ 48.68万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10734386
关键词：
1 year old 14-3-3 Family Address Age Animal Model Apnea Arousal Autopsy Back to Sleep Bioinformatics Biological Biological Markers Birth Blood Blood Platelets Blood Proteins Brain Brain Stem Cardiac Cause of Death Cessation of life Circulation Clinical Collection Control Groups Data Data Set Development Dryness Generations Glycoproteins Goals Hearing Immunohistochemistry Infant Intervention Investigation Laboratories Life Live Birth Metabolic Diseases Methodology Modeling Molecular Abnormality Multiple Abnormalities National Institute of Child Health and Human Development Neonatal Neonatal Screening Newborn Infant Parents Pathogenesis Pathway interactions Performance Peripheral Phenotype Prognostic Marker Proteins Proteome Proteomics ROC Curve Recording of previous events Reporting Research Risk Serotonin Serum Signaling Protein Sleep Smoking Spottings Stress Sudden infant death syndrome Techniques Testing Time Tissue Sample Tissues United States Western Blotting brain tissue comparison control critical developmental period critical period experimental study high risk infant infant death interest liquid chromatography mass spectrometry neonatal period neonate neurotransmission novel postneonatal mortality potential biomarker prognostic protein expression sample collection sex stressor validation studies

项目摘要

PROJECT SUMMARY Despite fewer deaths as a result of the Safe Sleep campaign, the Sudden Infant Death Syndrome (SIDS) remains the leading cause of post-neonatal infant mortality in the United States (0.33/1000 live births). Our research strongly implicates biological abnormalities within the brainstem that underlie an infant's vulnerability to SIDS. We have also identified SIDS-associated abnormalities within the serum and platelets taken from SIDS victims at autopsy. Together, these data suggest molecular abnormalities in the blood and brain of SIDS infants at death. What is unknown, however, is to what extent abnormalities observed at autopsy are also present at birth, and whether these or other, yet to be discovered, abnormalities can identify SIDS risk during the neonatal period, thereby providing the opportunity for intervention. Although there are no known biomarkers for SIDS risk, reports of apnea, cardiac rate abnormalities, and arousal deficits in infants who subsequently die of SIDS support the possibility that an underlying vulnerability may exist in the infant subclinically. Such subclinical abnormalities then manifest themselves when the infant is sufficiently stressed during a critical developmental period. Our laboratory has now acquired a unique dataset of dried blood spots (DBS) from neonatal screening cards from infants who later died of SIDS (n=109), died of known causes of death (demise controls, n=34), and age- and sex-matched control infants, identified as healthy at birth based on newborn screening for metabolic disorders (healthy controls (n=60)). In addition, we have corresponding brainstem tissue from autopsy in ~50% of these SIDS and demise control infants. Utilizing this dataset, we hypothesize that infants dying of SIDS display at birth abnormal protein profiles in DBSs compared to controls. Using liquid chromatography/mass spectrometry (LC/MS), we will test this hypothesis by performing an unbiased proteomic analysis of DBSs from SIDS and control infants (Specific Aim 1). Using the matching brainstem tissue we will use targeted methodologies, such as Western blotting, immunohistochemistry, and/or RNAscope to address the question of whether putative blood protein abnormalities at birth are represented in brain tissue collected at autopsy (Specific Aim 2). The use of neonatal DBSs for unbiased proteomics in SIDS is novel and the data generated from this unprecedented sample collection has potential to identify yet unknown proteins and/or pathways that play a role in SIDS pathogenesis. Furthermore, these data will allow us to assess identified abnormalities for their predictive potential for SIDS death and therefore their potential as neonatal biomarkers of SIDS risk. Given our overall goal, this application is fully aligned with NICHD's recent notice of special interest (NOT-HD-21-032) focused on identifying the root cause of SIDS with the aim to reduce SIDS-related deaths. The data to be generated from this R21 will provide novel prognostic information about SIDS infants at birth which will then be utilized as pilot data to expand upon in an R01 with new hypotheses, larger newborn datasets, and animal models.

项目概要尽管安全睡眠运动导致死亡人数减少，但婴儿猝死综合症 (SIDS) 仍然存在美国新生儿后婴儿死亡的主要原因（0.33/1000 活产婴儿）。我们的研究强烈暗示脑干内的生物异常是婴儿易患 SIDS 的基础。我们还在从 SIDS 受害者身上采集的血清和血小板中发现了与 SIDS 相关的异常在尸检时。总之，这些数据表明 SIDS 婴儿死亡时血液和大脑中存在分子异常。然而，尚不清楚尸检中观察到的异常在多大程度上也存在于出生时，以及无论是这些还是其他尚未发现的异常现象都可以识别新生儿时期的 SIDS 风险，从而提供干预的机会。尽管没有已知的 SIDS 风险生物标志物，但报告称随后死于 SIDS 的婴儿出现呼吸暂停、心率异常和觉醒缺陷的情况支持婴儿亚临床上可能存在潜在脆弱性的可能性。那么这种亚临床异常当婴儿在关键发育时期受到足够的压力时，就会表现出来。我们的实验室现在已经从婴儿的新生儿筛查卡中获得了干血斑 (DBS) 的独特数据集后来死于 SIDS (n=109)，死于已知死因（死亡对照，n=34），并且年龄和性别匹配对照婴儿，根据新生儿代谢紊乱筛查确定出生时健康（健康对照 (n=60))。此外，我们还从尸检中获得了约 50% 的 SIDS 和死亡患者的相应脑干组织控制婴儿。利用该数据集，我们假设死于 SIDS 的婴儿在出生时表现出异常蛋白质 DBS 中的概况与对照进行比较。使用液相色谱/质谱 (LC/MS)，我们将测试通过对 SIDS 和对照婴儿的 DBS 进行公正的蛋白质组学分析，证实了这一假设（具体目标1）。使用匹配的脑干组织，我们将使用有针对性的方法，例如蛋白质印迹法，免疫组织化学和/或 RNAscope 来解决假定的血液蛋白是否存在的问题尸检时收集的脑组织反映了出生时的异常情况（具体目标 2）。新生儿使用用于 SIDS 中无偏见蛋白质组学的 DBS 是新颖的，并且从这个前所未有的样本中生成的数据收集的数据有可能识别出在 SIDS 发病机制中发挥作用的未知蛋白质和/或途径。此外，这些数据将使我们能够评估已识别的异常情况，以预测 SIDS 的潜力死亡及其作为 SIDS 风险新生儿生物标志物的潜力。鉴于我们的总体目标，该应用程序与 NICHD 最近的特别关注通知 (NOT-HD-21-032) 完全一致，重点是识别根旨在减少 SIDS 相关死亡。从该 R21 生成的数据将提供有关出生时 SIDS 婴儿的新预后信息将用作试点数据来扩展 R01 中包含新假设、更大的新生儿数据集和动物模型。