Chronic hypoxia, AMPK activation and uterine artery blood flow

慢性缺氧、AMPK 激活与子宫动脉血流

基本信息

批准号：
9327023
负责人：
LORNA G. MOORE
金额：
$ 32.27万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-08-01 至 2021-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9327023
关键词：
Adenosine Monophosphate Affect Agonist Altitude Arteries Biometry Biopsy Blood Vessels Blood flow Blood specimen Caliber Catalytic Domain Cesarean section Chronic DNA Methylation Data Discipline of obstetrics Enzymes Epigenetic Process Fetal Growth Fetal Growth Retardation Fetal Tissues Frequencies Gene Expression Gene Expression Regulation Genes Genetic Transcription Growth Homeostasis House mice Human Hypoxia Infant Intervention Ischemia Link Measures Mediating Metabolic Metformin Methylation Mus Myometrial NIH Program Announcements Peripheral Blood Mononuclear Cell Pharmacology Phosphotransferases Placenta Plasma Play Positioning Attribute Postpartum Period Pre-Eclampsia Pregnancy Process Prospective Studies Protein Isoforms Protein Kinase Proteins Protocols documentation Public Health Publications Recruitment Activity Regulation Risk Role Sea Signal Pathway Signal Transduction Signaling Protein Single Nucleotide Polymorphism Testing Thoracic aorta Time Tissues Uteroplacental Circulation Vascular Smooth Muscle Vasodilation Vasodilator Agents Weight Western Blotting Woman Work cohort drug development epigenetic regulation fetal functional outcomes health of the mother human data improved in utero inhibitor/antagonist innovation mRNA Expression mouse model novel pregnancy disorder pregnant prevent residence response upstream kinase

项目摘要

Project Summary High altitude residence (HA, >2500 m) increases the frequency of preeclampsia (PreE) and intrauterine growth restriction (IUGR) 3-fold. Since hypoxia is a common cause, HA studies are uniquely positioned to evaluate the mechanistic role of hypoxia, discover new treatments, and alleviate the “almost complete lack of drug development for obstetric indications” noted in PAR-13-389. Adenosine monophosphate kinase (AMPK) is a regulator of metabolic homeostasis that also affects vascular growth and function under hypoxia. We have shown that AMPK plays important roles in the regulation of human uterine artery (UtA) blood flow and fetal growth at HA, and that AMPK activation has vasodilator effects in isolated murine UtA that are potentiated by hypoxia. Our central hypothesis is that AMPK activation promotes vasodilation in the uteroplacental circulation in response to hypoxia to raise UtA blood flow and improve fetal growth at HA. Because hypoxia leads to epigenetic changes that alter the expression of genes regulating of AMPK activity and fetal growth, we propose that DNA methylation influences these AMPK-mediated processes. We present new murine and human data to show that hypoxia a) raises placental phosphorylated (P)- relative to total AMPK levels and the P- as well as the total protein levels of AMPK targets in thoracic aorta and placenta; b) increases the expression of key enzymes activating AMPK in UtA and placenta; and c) alters methylation-expression relationships of AMPK- signaling genes important for fetal growth. We will test our central hypothesis by determining: 1. In Aim 1, the effect of HA pregnancy on AMPK signaling and its relationship to UtA blood flow and fetal growth in humans. We will recruit 102 healthy residents of low altitude (LA, 1600 m, n=53) or HA (3000 m, n=49); measure UtA blood flow and fetal biometry longitudinally; and determine the activation of AMPK, its upstream regulators and downstream targets, the expression levels and DNA methylation of relevant genes in peripheral blood mononuclear cells (PBMCs), and plasma levels of AMPK regulators. 2. In Aim 2, the effects of hypoxia on AMPK activation in human myometrial artery (MA) and placenta, and on MA vasoreactivity. In women participating in Aim 1 who deliver by elective C-section (n=19/altitude), we will obtain myometrial biopsies and placentas to determine a) the activation, expression and DNA methylation status of AMPK, its well-established regulators and downstream targets; b) the effect of HA pregnancy on MA vasoreactivity; and c) whether hypoxia potentiates vasodilator effects of AMPK activation in MA. 3. In Aim 3, the role of AMPK in regulating uteroplacental blood flow and fetal growth in response to hypoxia in mice. Pregnant mice housed at sea level (SL) or HA will be treated with the AMPK activator (AICAR), inhibitor (Compound C), or vehicle (control) for determining the separate and combined effects of hypoxia and AMPK activation on AMPK signaling in UtA, placental and fetal tissues; UtA vascular reactivity and blood flow; and fetal growth.

项目概要高海拔居住（HA，>2500 m）会增加先兆子痫（PreE）和宫内生长的频率限制 (IUGR) 3 倍由于缺氧是常见原因，HA 研究具有独特的地位来评估缺氧。缺氧的机制作用，发现新的治疗方法，缓解“几乎完全缺药” PAR-13-389 中指出的“产科适应症的开发”是一种单磷酸腺苷激酶 (AMPK)。代谢稳态的调节剂，也会影响缺氧条件下的血管生长和功能。表明 AMPK 在调节人子宫动脉 (UtA) 血流和胎儿发育中发挥重要作用 HA 的生长，并且 AMPK 激活对分离的鼠 UtA 具有血管舒张作用，这种作用通过我们的中心假设是 AMPK 激活促进子宫胎盘循环中的血管舒张。应对缺氧以增加 UtA 血流量并改善 HA 处的胎儿生长。我们建议表观遗传变化会改变调节 AMPK 活性和胎儿生长的基因表达 DNA 甲基化影响这些 AMPK 介导的过程，我们向其提供了新的小鼠和人类数据。表明缺氧 a) 会提高胎盘磷酸化 (P)- 相对于总 AMPK 水平以及 P- 和 b) 胸主动脉和胎盘中 AMPK 靶标的总蛋白水平增加；激活 UtA 和胎盘中 AMPK 的酶；以及 c) 改变 AMPK- 的甲基化表达关系我们将通过确定以下内容来检验我们的中心假设： 1. 目标1，HA妊娠对AMPK信号传导的影响及其与UtA血流和胎儿的关系我们将招募 102 名低海拔（LA，1600 m，n=53）或 HA（3000 m， n=49);纵向测量UtA血流和胎儿生物特征并确定AMPK的激活；上游调控因子和下游靶标，表达水平与DNA甲基化的相关性外周血单核细胞 (PBMC) 中的基因以及 AMPK 调节因子的血浆水平。 2. 在目标 2 中，缺氧对人子宫肌动脉 (MA) 和胎盘中 AMPK 激活的影响，以及 MA 血管反应性对于参与目标 1 并通过选择性剖腹产分娩的女性（n=19/海拔高度），我们将获取子宫肌层活检和胎盘以确定 a) 激活、表达和 DNA 甲基化 AMPK 的状态、其完善的调节因子和下游靶标 b) HA 妊娠对胎儿的影响； MA 血管反应性；c) 缺氧是否增强 MA 中 AMPK 激活的血管舒张作用。 3. 目标3中，AMPK在缺氧反应中调节子宫胎盘血流和胎儿生长的作用饲养在海平面 (SL) 或 HA 的怀孕小鼠将接受 AMPK 激活剂 (AICAR) 治疗，抑制剂（化合物 C）或载体（对照），用于确定缺氧的单独和组合效应 AMPK 激活对 UtA、胎盘和胎儿组织中 AMPK 信号传导的影响；血流量；和胎儿生长。