Role of E2B Metabolites in Uterine Artery Endothelial Adaptations to Pregnancy

 描述(由申请人提供)：这份F31-多样性申请表为Rosalina Villalon Landeros提供了一份详细的培训计划，以实现她完成的职业生涯目标 她的博士学位，过渡到一个高水平的博士后职位，并成为一名独立研究员。她的培训计划确定了指导和培训的领域，例如：撰写手稿和助学金，2)专业网络，3)教学/指导和领导力。她的科学项目由她的赞助商(RR Magness)NIH拨款5P01HD038843-12和其他NIH拨款资助，这些拨款涉及子宫动脉对怀孕的适应以及就近的局部环境与系统荷尔蒙状况在这种适应中的作用。子宫动脉内皮细胞(UAECs)在子宫血管适应机制中发挥核心作用，包括血管生成和血管扩张，这是增加正常妊娠期间子宫血流量(UBF)所必需的。子宫-胎盘血管适应异常导致的母胎循环不足导致胎儿发育不良 宫内生长受限(IUGR)。雌二醇(E_2，β)及其代谢产物--儿茶酚-雌二醇2-和4-羟基雌二醇，以及甲氧基雌二醇2-甲氧基雌二醇和4-甲氧基雌二醇，在正常妊娠期间通过促进子宫动脉内皮细胞的增殖、前列环素和一氧化氮的产生而调节子宫动脉的功能，从而促进血管生成和血管扩张。我们的实验室建立了一种独特的绵羊手术诱导单侧妊娠模型，通过该模型，我们发现与对侧动脉相比，妊娠角同侧的子宫动脉经历了实质性的重塑。该模型将使我们能够比较局部妊娠环境和全身激素环境对子宫动脉适应性和内皮细胞编程的影响。总的假设是，子宫动脉妊娠角同侧和对侧将表现出不同的编程差异，使从同侧子宫动脉分离的UAECs[(妊娠)P-UAECs]将表现出与正常妊娠的P-UAECs相同的编程(增加血管生成和血管扩张)，而从妊娠角对侧子宫动脉分离的UAECs[(非妊娠)P-UAECs]将表现出与骑自行车羊的NP-UAECs相同的编程(低血管生成和血管扩张剂产生)。将使用(妊娠期)P-UAECs、(非妊娠期)P-UAECs和P-UAECs NP-UAECs，这些细胞以前是从健康母羊身上分离和验证的，无论是否进行了子宫手术修改。通过检测E2β及其代谢物刺激的血管生成(通过增殖分析)和血管扩张剂的产生(通过免疫分析和高效液相色谱)，以及与正常和非妊娠的比较，将确定妊娠角同侧和对侧UAECs的编程差异。这些研究将促进我们对子宫血管对妊娠适应性的理解，并为未来的研究奠定平台，旨在确定局部妊娠环境中分泌的因素，这些因素可能导致IUGR和先兆子痫的预防或纠正治疗的创新。

 DESCRIPTION (provided by applicant): This F31-Diversity application provides a detailed training plan for Rosalina Villalon Landeros to fulfill her professional career goals of completing her PhD degree, transition into a high level postdoc position, and become an independent researcher. Her training plan identifies areas of mentorship and training such as: writing manuscripts and grants, 2) professional networking, 3) Teaching/Mentorship and leadership. Her scientific project is funded by her sponsor's (RR Magness) NIH grant 5P01HD038843-12 and other NIH grants that relate to uterine artery adaptations to pregnancy and the role of the immediate local environment versus the systemic hormonal profile on such adaptations. Uterine artery endothelial cells (UAECs) play a central role in the mechanisms underlying uterine vascular adaptations, including angiogenesis and vasodilatation, which are necessary to increase uterine blood flow (UBF) during normal pregnancy. Inadequate maternal-fetal circulation as a result of aberrant utero-placental vascular adaptations results in decreased fetal growth and ultimately in intrauterine growth restriction (IUGR). Estradiol (E2β) and its metabolites, the catecholestradiols 2-OHE2 and 4-OHE2 and the methoxyestradiols 2-ME2 and 4-ME2, are known to regulate uterine artery function during normal pregnancy by increasing UAEC proliferation, prostacyclin and nitric oxide production resulting in increased angiogenesis and vasodilatation. Our laboratory developed a unique ovine surgical-induced unilateral pregnancy model, through which we have found that uterine arteries ipsilateral to the gravid horn undergo substantial remodeling compared to contralateral arteries. This model will allow us to compare the effects of the local gravid environment to the systemic hormonal milieu on uterine artery adaptation and endothelial cell programming. The overall hypothesis is that uterine arteries ipsilateral and contralateral to the gravid horn will exhibit differences in programming so that UAECs isolated from uterine arteries ipsilateral to the gravid horn [(Gravid)P-UAECs] will exhibit the same programming (increased angiogenesis and vasodilation) as P-UAECs of normal pregnancies, while UAECs isolated from uterine arteries contralateral to the gravid horn [(NonGravid)P-UAECs] will exhibit the same programing (lower angiogenesis and vasodilator production) as NP-UAECs of cycling ewes. (Gravid)P-UAECs, (NonGravid)P- UAECs, and P-UAECs NP-UAECs, previously isolated and validated from healthy ewes with or without uterine surgical modification will be used. Differences in programming of UAECs ipsilateral and contralateral to the gravid horn will be determined by examining angiogenesis (through proliferation assays) and vasodilator production (through immunoassays and HPLC) stimulated by E2β and its metabolites and in comparison to normal and nonpregnancy. These studies will advance our understanding of uterine vascular adaptations to pregnancy and set the platform for future research aimed at identifying factors secreted in the local gravid milieu which may lead to the innovation of preventive or corrective treatments for IUGR and preeclampsia.