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

• 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
• 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

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）：宫内生长限制（IUGR）增加了成人疾病的围产期并发症和易感性的风险。但是，介导的长期健康增长和编程的机制尚待充分确定。我们以前提供了令人信服的证据，表明雷帕霉素（MTOR）信号的滋养细胞哺乳动物靶标充当胎盘营养传感器，通过调节胎盘营养转运蛋白来间接影响胎儿生长。在此提案中，我们将提出创新的假设，即胎盘MTOR信号直接影响胎儿代谢和生长。我们的核心假设是，响应限制营养物的可用性抑制滋养细胞MTOR信号传导会改变释放的体液因子的释放，从而导致IGFBP-1的分泌和磷酸化增加，而IGF-I从胎儿肝脏中的分泌减少。我们提出了两个具体目的：目标1：确定滋养细胞MTOR在IGF-I和IGFBP-1分泌的调节中的作用，以及IGFBP-1磷酸化在胎儿肝细胞中的体外和目标2：确定胎盘属MTOR对胎儿特异性MTOR对胎儿liver liver liver IGF-I和IGFBP-1分泌的影响。在AIM 1中，我们将从两种类型的人类原代滋养细胞细胞中获得条件培养基（CM）：1。从正常项胎盘中分离出来，并用靶向MTOR信号途径的crampled siRNA或siRNA转染的细胞和2个。与IUGR分离的2个细胞。人类HEPG2细胞和原发性胎儿狒狒肝细胞将在滋养细胞CM中孵育，并确定IGFBP-1（Western blot，2-D免疫印迹，ELISA和质谱法）的分泌和磷酸化以及IGF-I分泌（ELISA）。此外，将使用基于质谱的定量标记方法来确定推定的体液因子的候选物。在AIM 2中，我们将使用含有ERT2-Cre-Cronctucts的慢病毒载体从MTOR-串联小鼠中转染MTOR-串联小鼠的胚泡细胞来开发有条件的滋养细胞MTOR敲击小鼠。 IGFBP-1的表达和磷酸化和IGF-I在胎儿肝脏中的表达和IGF-1水平的IGF-I和IGFBP-1水平以及胎儿生长以及胎儿的生长将在诱导培养基细胞特异性MTOR击倒后，研究。意义：异常的胎儿生长是导致围产期发病率的主要因素，并对长期健康产生了深远的影响。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，并在以后的生活中为疾病编程。创新：假设胎盘通过调节胎儿肝功能直接调节胎儿生长是一个概念上的新思想。此外，我们建议开发一种方法，允许小鼠中有条件的滋养细胞特异性基因靶向，这是我们所知的 - 以前没有报道过。因此，拟议的工作在方法论上也是创新的。

项目成果

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

• DOI: 10.1016/j.placenta.2012.05.010
• 发表时间: 2012-11
• 期刊: PLACENTA
• 影响因子: 3.8
• 作者: Jansson, T.;Aye, I. L. M. H.;Goberdhan, D. C. I.
• 通讯作者: Goberdhan, D. C. I.

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

• DOI: 10.3390/ijms24087273
• 发表时间: 2023-04-14
• 期刊: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
• 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