Molecular Mechanisms Linking Placental Nutrient Sensing and Fetal Programming

连接胎盘营养感应和胎儿编程的分子机制

基本信息

批准号：
8432440
负责人：
LESLIE MYATT
金额：
$ 16.21万
依托单位：
UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2015-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8432440
关键词：
Adult Affect Affinity Amino Acid Transporter Amino Acids Beta Cell Binding Brain Cardiovascular Diseases Cell physiology Cell secretion Cells Childhood Conditioned Culture Media Coupled Data Diabetes Mellitus Disease Drosophila genus Endocrine Enzyme-Linked Immunosorbent Assay Epigenetic Process Fat Body Fetal Growth Fetal Growth Retardation Fetal Liver Fetal Tissues Fetal Weight Fetus Functional disorder Gene Targeting Genetic Transcription Gestational Age Growth Growth Factor Health Hepatocyte Hormones Human Hypoxia Immunoblotting In Vitro Incubated Injury Insulin-Like Growth Factor I Insulin-Like Growth-Factor Binding Protein 1 Knockout Mice Label Lentivirus Vector Life Link Liquid Chromatography Liver Mass Spectrum Analysis Mediating Metabolic Metabolism Modeling Molecular Morbidity - disease rate Mus Muscle function Nutrient Obesity Oxidative Stress Oxygen Pancreas Papio Peptides Perinatal Phosphorylation Placenta Play Proteins Regulation Reporting Research Risk Role Serum Signal Pathway Signal Transduction Skeletal Muscle Small Interfering RNA Source Tamoxifen Testing Tetanus Helper Peptide Time Tissues Transfection Translations Western Blotting Work base blastocyst cell growth design detection of nutrient fetal fetal programming fetus cell in vivo innovation insulin secretion knock-down liver function mTOR protein novel programs receptor release factor response sensor trophoblast

项目摘要

DESCRIPTION (provided by applicant): Intrauterine growth restriction (IUGR) increases the risk for perinatal complications and predisposes for adult disease. However, the mechanisms mediating the restricted growth and programming of long-term health remain to be fully established. We have previously provided compelling evidence that trophoblast mammalian target of rapamycin (mTOR) signaling functions as a placental nutrient sensor, indirectly influencing fetal growth by regulating placental nutrient transporters. In this proposal we will tet the innovative hypothesis that placental mTOR signaling directly influences fetal metabolism and growth. Our central hypothesis is that inhibition of trophoblast mTOR signaling in response to restricted nutrient availability alters the release of humoral factors that cause increased secretion and phosphorylation of IGFBP-1 and decreased secretion of IGF-I from the fetal liver. We propose two specific aims: Aim 1: Determine the role of trophoblast mTOR in the regulation of IGF-I and IGFBP-1 secretion and IGFBP-1 phosphorylation in fetal liver cells in vitro and Aim 2: Establish the effect of placental specific mTOR knock down on fetal liver IGF-I and IGFBP-1 secretion and IGFBP-1 phosphorylation in the mouse. In Aim 1, we will obtain conditioned media (CM) from cultured human primary trophoblast cells of two types: 1. Cells isolated from normal term placentas and transfected with scrambled siRNA or siRNA targeting the mTOR signaling pathway and 2. Cells isolated from IUGR and Appropriate-for-Gestational-Age (AGA) placentas. Human HepG2 cells and primary fetal baboon liver cells will be incubated in trophoblast CM and secretion and phosphorylation of IGFBP-1 (Western blot, 2-D immunoblotting, ELISA and mass spectrometry approaches) as well as IGF-I secretion (ELISA) will be determined. In addition, candidates for the putative humoral factors will be identified using mass spectrometry based quantitative labeling approaches. In Aim 2, we will develop a conditional trophoblast specific mTOR knock down mouse using transfection of blastocysts from mTOR-floxed mice by lentiviral vectors containing ERT2-Cre-constructs. The expression and phosphorylation of IGFBP-1 and expression of IGF-I in fetal liver and fetal levels of IGF-I and IGFBP-1 as well as fetal growth will be studied after induction of trophoblast specific mTOR knock down by tamoxifen. Significance: Abnormal fetal growth is a major contributor to perinatal morbidity and has profound impact on long-term health. This work has the potential to identify a molecular mechanism by which the placenta directly influences fetal metabolism and growth and programs the fetus for disease later in life. Innovation: The hypothesis that the placenta directly regulates fetal growth by modulating fetal liver function is a conceptually novel idea. Furthermore, we propose to develop an approach allowing conditional trophoblast specific gene targeting in the mouse, which has - to the best of our knowledge - not previously been reported. Thus, the proposed work is also methodologically innovative.

描述（由申请人提供）：宫内生长受限（IUGR）会增加围产期并发症的风险并容易患成人疾病。然而，调节增长受限和长期健康规划的机制仍有待充分建立。我们之前提供了令人信服的证据，证明滋养层哺乳动物雷帕霉素靶标（mTOR）信号传导可作为胎盘营养传感器，通过调节胎盘营养转运蛋白间接影响胎儿生长。在本提案中，我们将提出一个创新假设：胎盘 mTOR 信号传导直接影响胎儿的代谢和生长。我们的中心假设是，因营养供应受限而抑制滋养层 mTOR 信号传导会改变体液因子的释放，从而导致 IGFBP-1 的分泌和磷酸化增加，并减少胎儿肝脏中 IGF-I 的分泌。我们提出了两个具体目标：目标 1：确定滋养层 mTOR 在体外调节胎儿肝细胞中 IGF-I 和 IGFBP-1 分泌以及 IGFBP-1 磷酸化中的作用；目标 2：建立胎盘特异性 mTOR 敲除的效果降低胎儿肝脏 IGF-I 和 IGFBP-1 分泌以及小鼠 IGFBP-1 磷酸化。在目标 1 中，我们将从培养的两种类型的人原代滋养层细胞中获得条件培养基 (CM)：1. 从正常足月胎盘分离并用乱序 siRNA 或靶向 mTOR 信号通路的 siRNA 转染的细胞，以及 2. 从 IUGR 和 IUGR 分离的细胞适合孕龄 (AGA) 胎盘。人 HepG2 细胞和原代胎儿狒狒肝细胞将在滋养层 CM 中孵育，IGFBP-1 的分泌和磷酸化（蛋白质印迹、2-D 免疫印迹、ELISA 和质谱方法）以及 IGF-I 分泌 (ELISA) 将被检测。决定。此外，将使用基于质谱的定量标记方法来鉴定推定体液因子的候选者。在目标 2 中，我们将通过含有 ERT2-Cre 构建体的慢病毒载体转染 mTOR-floxed 小鼠的囊胚，开发条件性滋养层特异性 mTOR 敲低小鼠。在用他莫昔芬诱导滋养层特异性mTOR敲低后，将研究IGFBP-1的表达和磷酸化以及胎儿肝脏中IGF-I的表达以及胎儿IGF-I和IGFBP-1的水平以及胎儿生长。意义：胎儿生长异常是围产期发病的主要原因，并对长期健康产生深远影响。这项工作有可能确定胎盘直接影响胎儿代谢和生长的分子机制，并为胎儿在以后的生活中预防疾病做好准备。创新：胎盘通过调节胎儿肝功能直接调节胎儿生长的假设在概念上是一个新颖的想法。此外，我们建议开发一种方法，允许在小鼠中进行条件性滋养层特异性基因靶向，据我们所知，此前尚未有报道。因此，拟议的工作在方法上也是创新的。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The role and regulation of IGFBP-1 phosphorylation in fetal growth restriction.

IGFBP-1磷酸化在胎儿生长受限中的作用和调节。

DOI：
发表时间：
2015-06
期刊：
影响因子：
0
作者：
Gupta; Madhulika B
通讯作者：
Madhulika B

Placental Remote Control of Fetal Metabolism: Trophoblast mTOR Signaling Regulates Liver IGFBP-1 Phosphorylation and IGF-1 Bioavailability.

胎盘远程控制胎儿代谢：滋养层 mTOR 信号传导调节肝脏 IGFBP-1 磷酸化和 IGF-1 生物利用度。

DOI：
发表时间：
2023-04-14
期刊：
影响因子：
5.6
作者：
Rosario, Fredrick J;Chopra, Anand;Biggar, Kyle;Powell, Theresa L;Gupta, Madhulika B;Jansson, Thomas
通讯作者：
Jansson, Thomas

The emerging role of mTORC1 signaling in placental nutrient-sensing.

mTORC1 信号在胎盘营养感应中的新作用。