Mechanisms Modulating the Association between the ENG Pathway and Preeclampsia

ENG 通路与先兆子痫之间关联的调节机制

基本信息

批准号：
8591807
负责人：
Mandy Jo Schmella
金额：
$ 4.98万
依托单位：
MAGEE-WOMEN'S RES INST AND FOUNDATION
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-01 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8591807
关键词：
Address Affect African American American Angiogenic Factor Animal Model Attenuated Base Sequence Biological Assay Blood Blood Circulation Blood Vessels Blood capillaries Calcitriol California Candidate Disease Gene Caucasians Caucasoid Race Cell Culture Techniques Cell Line Cells Cholecalciferol Classification Clinical Collection DNA Detection Discrimination Disease Doctor of Philosophy Endoglin Endothelial Cells Enzyme-Linked Immunosorbent Assay Expenditure Family Functional disorder Funding Genetic Genetic Variation Genomics Genotype Gold HELLP Syndrome Health Healthcare Human Hypoxia Individual Intervention Investigation Knowledge Maternal Age Measurement Measures Messenger RNA Metabolic Methods Mission Morbidity - disease rate National Institute of Nursing Research Neonatal Nitric Oxide Norway Outcome Output Oxygen Pathway interactions Patients Phenotype Plasma Population Pre-Eclampsia Predisposition Pregnancy Pregnancy Outcome Pregnancy Proteins Prevention Preventive Proteins Reporting Research Risk Rodent Role Sampling Scheme Science Serum Single Nucleotide Polymorphism Spiral Artery of the Endometrium Study Subject Susceptibility Gene Syndrome System Testing Therapeutic Time Transforming Growth Factor beta Receptors Translating University Hospitals Up-Regulation Validation Variant Vitamin D Vitamin D Response Element Western Blotting Woman base biobank capillary caucasian American design evidence base genetic association genetic variant implantation improved insight mRNA Expression mortality neonatal morbidity parity pregnancy disorder pregnant prevent promoter public health relevance research study treatment strategy trophoblast

项目摘要

DESCRIPTION (provided by applicant): The pathophysiology of preeclampsia (PE) is unclear, and effective strategies/treatments to successfully reduce global maternal and neonatal morbidity/mortality are lacking. The long term objective of this project is to improve our understanding of the biologic underpinnings of PE so that effective prevention, detection, and treatment interventions can be developed to identify women at risk and improve health outcomes of moms and babies affected by PE. This project aims to address significant gaps in our understanding of PE pathophysiology by focusing on improving our understanding of endoglin (ENG) pathway dysregulation in PE. We know that elevations in circulating soluble endoglin (sENG) protein and placental/blood ENG mRNA expression precede clinical presentation of PE, and increased levels of sENG in rodents produce a PE-like syndrome that can be amplified by other anti-angiogenic factors. I have previously demonstrated that genetic variation in the ENG pathway is associated with PE in American Caucasian and African American samples. However, it is unknown if genetic variability in the ENG pathway impacts ENG mRNA expression and/or sENG levels, or if metabolic factors (e.g., Vitamin D down in PE) modulate cellular sENG output. This project addresses mechanisms of endoglin pathway dysregulation and validation of our previous genetic findings, both of which are necessary to provide the evidence based needed to eventually translate these findings clinically. First, using a candidate gene case-control design, we will test the hypothesis that our genetic association findings are present in other populations from the Bill & Melinda Gates funded Global Pregnancy CoLaboratory, a biobank consortium of >20 research groups studying adverse pregnancy outcomes. Second, we will conduct focused genotyping (missense single nucleotide polymorphism assessment; sequencing) of ENG to identify common and/or rare functional variants that are involved in PE susceptibility, using a case-control design. Third, we will explor mechanisms underlying associations between the ENG pathway: (a) test the relationship of pregnancy plasma levels (e.g., sENG) to ENG pathway genotypes (within-case OR within-control design) and pregnancy outcome (case-control design); (b) test the hypothesis that Vitamin D attenuates sENG release from human trophoblast and uterine endothelial cell lines cultured under basal and stimulated (hypoxia, PE serum) conditions. Methods to achieve these specific aims, germane to the National Institute of Nursing Research's mission to promote health and prevent disease through identification of susceptibility genes for at- risk individuals, includ genotype collection with either the i-PLEX(R) Gold SNP assay or TaqMan(R) allelic discrimination, capillary based sequencing, protein measurement with ELISA assays and Western Blot, and cell culture under varying treatment conditions (e.g., oxygen content, Vitamin D concentration, patient serum). Ultimately, results from this project will improve our understanding of the pathophysiologic underpinnings of PE and may help to identify preventive, predictive, and therapeutic/modifiable targets for women at risk for PE.

描述（由申请人提供）：先兆子痫（PE）的病理生理学尚不清楚，并且缺乏成功降低全球孕产妇和新生儿发病率/死亡率的有效策略/治疗。该项目的长期目标是提高我们对早泄的生物学基础的了解，以便制定有效的预防、检测和治疗干预措施，以识别处于危险中的妇女并改善受早泄影响的母亲和婴儿的健康状况。该项目旨在通过重点提高我们对 PE 中内皮糖蛋白 (ENG) 通路失调的理解，弥补我们对 PE 病理生理学理解上的重大差距。我们知道，循环可溶性内皮糖蛋白 (sENG) 蛋白和胎盘/血液 ENG mRNA 表达的升高先于 PE 的临床表现，并且啮齿类动物中 sENG 水平的升高会产生类似 PE 的综合征，而其他抗血管生成因子可以放大这种综合征。我之前已经证明，ENG 通路中的遗传变异与美国白种人和非裔美国人样本中的 PE 相关。然而，尚不清楚 ENG 通路的遗传变异是否影响 ENG mRNA 表达和/或 sENG 水平，或者代谢因素（例如 PE 中的维生素 D 下降）是否调节细胞 sENG 输出。该项目解决了内皮糖蛋白通路失调的机制并验证了我们之前的遗传发现，这两者都是提供最终将这些发现转化为临床所需的证据所必需的。首先，使用候选基因病例对照设计，我们将检验以下假设：我们的遗传关联发现存在于比尔和梅琳达·盖茨资助的全球妊娠联合实验室的其他人群中，该实验室是一个由超过 20 个研究小组组成的生物银行联盟，研究不良妊娠结局。其次，我们将使用病例对照设计对 ENG 进行集中基因分型（错义单核苷酸多态性评估；测序），以识别与 PE 易感性相关的常见和/或罕见功能变异。第三，我们将探索 ENG 通路之间关联的潜在机制：(a) 测试妊娠血浆水平（例如 sENG）与 ENG 通路基因型（病例内或对照内设计）和妊娠结局（病例对照设计）的关系）； (b) 检验维生素 D 减弱在基础和刺激（缺氧、PE 血清）条件下培养的人滋养层和子宫内皮细胞系的 sENG 释放的假设。实现这些具体目标的方法与国家护理研究所的使命密切相关，即通过识别高危个体的易感基因来促进健康和预防疾病，包括使用 i-PLEX(R) Gold SNP 测定或 TaqMan 进行基因型收集(R) 等位基因区分、基于毛细管的测序、使用 ELISA 测定和蛋白质印迹进行蛋白质测量，以及不同处理条件（例如氧含量、维生素 D 浓度、患者血清）下的细胞培养。最终，该项目的结果将提高我们对 PE 病理生理学基础的理解，并可能有助于为有 PE 风险的女性确定预防、预测和治疗/可修改的目标。