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 尽管由于安全睡眠运动而导致死亡人数较少，但婴儿突然死亡综合症（SIDS）仍然存在 美国昆虫后婴儿死亡率的主要原因（0.33/1000活产）。我们的研究 强烈暗示着脑干内的生物学异常，这是婴儿脆弱性小岛屿发展中国家的基础。 我们还确定了血清中与SIDS受害者的血清和血小板中与SID相关的异常 在尸检。总之，这些数据表明死亡时小岛屿发展中国家的血液和大脑的分子异常。 然而，未知的是在出生时也存在尸检时观察到的异常，并且 这些或其他尚未发现的这些异常是否可以识别在新生儿时期的SID风险， 从而提供干预的机会。虽然没有SIDS风险的已知生物标志物，但报道 呼吸暂停，心率异常和随后死亡的婴儿的唤醒缺陷支持 潜在的脆弱性可能在下属性中存在。这样的亚临床异常 当婴儿在关键发育时期充分压力时，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取了独特的数据集（DBS） 后来死于小岛屿发展中国家（n = 109），死于已知的死亡原因（Dimise Controls，n = 34），并与年龄和性别匹配 对照婴儿，基于新生儿筛查代谢疾病（健康对照），在出生时被认为是健康的 （n = 60））。此外，我们在这些SID和灭亡中有相应的脑干组织。 控制婴儿。利用该数据集，我们假设在出生异常蛋白质中垂死的婴儿死亡的婴儿 与对照组相比，DBSS中的轮廓。使用液相色谱/质谱法（LC/MS），我们将测试 通过对小岛屿发展中国家和对照婴儿的DBS进行公正的蛋白质组学分析（特定），该假设 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Western blotting， 免疫组织化学和/或rnascope，以解决假定血蛋白的问题 出生时异常在尸检时收集的脑组织（特定目标2）。新生儿的使用 SIDS中无偏蛋白质组学的DBS是新颖的，并且从此前所未有的样本中产生的数据 收集有可能识别尚未知道的蛋白质和/或途径，这些蛋白质和/或途径在SIDS发病机理中起作用。 此外，这些数据将使我们能够评估已确定的异常情况，以确定的SIDS的预测潜力 因此，死亡，因此作为SIDS新生儿生物标志物的潜力风险。鉴于我们的总体目标，这个应用程序 完全与NICHD最近的特殊兴趣通知（NOT-HD-21-032）完全一致，该通知旨在识别根源 小岛屿发展中国家的原因，目的是减少与SIDS相关的死亡。从此R21生成的数据将提供 关于小岛屿发展中国家出生时小岛婴儿的新型预后信息，然后将其用作试点数据来扩展 在带有新假设的R01中，较大的新生儿数据集和动物模型。