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

• 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
• 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

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.

项目摘要 尽管安全睡眠运动减少了死亡人数，但婴儿猝死综合症（SIDS）仍然存在。 这是美国新生儿后期婴儿死亡率的主要原因（0.33/1000活产）。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。 我们还在取自SIDS患者的血清和血小板中发现了与SIDS相关的异常 尸体解剖时。总之，这些数据表明SIDS婴儿死亡时血液和大脑中的分子异常。 然而，目前尚不清楚的是，在尸检中观察到的异常在多大程度上也存在于出生时， 无论是这些还是其他尚未发现的异常，都可以识别新生儿期SIDS的风险， 从而提供干预的机会。尽管目前还没有已知的SIDS风险的生物标志物， 在随后死于SIDS的婴儿中，呼吸暂停、心率异常和唤醒缺陷支持了 婴儿亚临床存在潜在脆弱性的可能性。这种亚临床异常， 当婴儿在关键的发育时期受到足够的压力时，就会表现出来。本实验室 现在已经从新生儿筛查卡上获得了一个独特的干血斑（DBS）数据集，这些新生儿筛查卡来自 后来死于SIDS（n=109），死于已知死因（死亡对照，n=34），年龄和性别匹配 对照婴儿，根据新生儿代谢紊乱筛查确定为出生时健康（健康对照 （n=60））。此外，我们在约50%的SIDS和死亡病例中的尸检中获得了相应的脑干组织。 控制婴儿。利用这个数据集，我们假设死于SIDS的婴儿在出生时显示异常蛋白质， 与对照相比，DBS中的特征。使用液相色谱/质谱（LC/MS），我们将测试 通过对来自SIDS和对照婴儿的DBS进行无偏蛋白质组学分析（特异性 目标1）。使用匹配的脑干组织，我们将使用靶向方法，如蛋白质印迹法， 免疫组织化学和/或RNA显微镜来解决假定的血液蛋白是否 出生时的异常表现在尸检时收集的脑组织中（具体目标2）。新生儿使用 DBS在SIDS中用于无偏蛋白质组学是新颖的， 收集有可能确定未知的蛋白质和/或途径发挥作用的SIDS发病机制。 此外，这些数据将使我们能够评估已确定的异常对SIDS的预测潜力 因此，它们有可能成为SIDS风险的新生儿生物标志物。考虑到我们的总体目标， 完全符合NICHD最近的特别关注通知（NOT-HD-21-032），重点是确定根源 小岛屿发展中国家的原因，以减少与小岛屿发展中国家有关的死亡。R21生成的数据将提供 有关SIDS婴儿出生时的新预后信息，然后将用作试点数据进行扩展 在R 01中使用新的假设，更大的新生儿数据集和动物模型。