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：
8731148
负责人：
Mandy Jo Schmella
金额：
$ 4.76万
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/8731148
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 rare variant 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）：尚不清楚先兆子痫（PE）的病理生理学，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/治疗方法。该项目的长期目标是提高我们对PE生物学基础的理解，以便可以开发出有效的预防，检测和治疗干预措施，以识别有风险的女性并改善受PE影响的妈妈和婴儿的健康成果。该项目的目的是通过重点提高我们对PE中的内元（ENG）途径失调的理解来解决我们对PE病理生理学的理解的显着差距。我们知道，循环可溶性内凝蛋白（SENG）蛋白和胎盘/血液ENG mRNA表达的升高先于PE的临床表现，而啮齿动物的Seng水平升高会产生一种PE样综合征，可以通过其他抗血管生成因子扩大。我以前已经证明，在美国高加索和非裔美国人样本中，ENG途径的遗传变异与PE有关。但是，尚不清楚ENG途径中的遗传变异性是否会影响ENG mRNA表达和/或SENG水平，或者代谢因子（例如，PE中的维生素D下降）是否会调节细胞SENG输出。该项目解决了内og途径失调的机制和我们以前的遗传发现的验证，这两者都是为了提供最终在临床上翻译这些发现所需的基于证据的必要条件。首先，使用候选基因病例对照设计，我们将检验以下假设：在Bill＆Melinda Gates的其他人群中，我们的遗传关联发现资助了全球妊娠合伙人，这是一个> 20个研究小组的生物库联盟，研究了不良妊娠成果。其次，我们将使用病例对照设计，进行ENG的集中基因分型（错过单核苷酸多态性评估；测序），以识别与PE敏感性有关的常见和/或稀有功能变体。第三，我们将探索ENG途径之间关联的基础机制：（a）测试怀孕血浆水平（例如Seng）与ENG途径基因型（案例内部或控制内部设计）和妊娠结局（病例控制设计）的关系；（b）检验以下假设：维生素D可减弱Seng从人类滋养细胞和子宫内皮细胞系中释放的假说，该细胞在基础和刺激（缺氧，PE血清）条件下培养的情况。 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含量，维生素D浓度，患者血清）。最终，该项目的结果将提高我们对PE的病理生理基础的理解，并可能有助于确定有PE风险的女性的预防，预测和治疗/可修改目标。