Role of Hypoxia in Regulating Stromal-Epithelial Communication during Pregnancy

妊娠期缺氧在调节间质-上皮通讯中的作用

基本信息

批准号：
10166891
负责人：
MILAN K BAGCHI
金额：
$ 30.78万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-13 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10166891
关键词：
Address Amino Acids Animal Model Biology Cell Communication Cell Culture Techniques Cell secretion Cells Data Decidual Cell Reactions Defect Distant Down-Regulation Endometrial Endometrial Stromal Cell Endometrium Environment Epithelial Epithelial Cells Epithelial-Stromal Communication Exhibits Genomics Goals Human Hypoxia Hypoxia Inducible Factor In Vitro Infertility Link Mediating Messenger RNA Metabolic MicroRNAs Modeling Molecular Mus Paracrine Communication Pathway interactions Pregnancy Process Proteins Proteomics Regulation Reporting Research Role Signal Transduction Signaling Molecule Stromal Cells Supporting Cell Testing Time Tissues Transcript Uterus bHLH-PAS factor HLF blastocyst cell type early pregnancy exosome experimental study extracellular vesicles failure Implantation fetal implantation in vivo insight intercellular communication metabolomics microvesicles mouse model natural Blastocyst Implantation novel paracrine rab GTP-Binding Proteins response trafficking transcription factor

项目摘要

Implantation is initiated when the blastocyst attaches to the uterine luminal epithelium and subsequently penetrates into the underlying stroma to firmly embed into the endometrium. While many aspects of implantation remain unclear, emerging evidence indicates that intercellular communication between endometrial epithelial and stromal cells is vital for successful establishment of pregnancy. The paracrine signals that mediate this crosstalk remain poorly understood. Our recent studies revealed that the transcription factor, hypoxia-‐inducible factor 2 alpha (HIF2α), is induced selectively in endometrial stromal cells subjacent to the luminal epithelium at the time of implantation. This finding is significant because it is known for a long time that the maternal environment during implantation is hypoxic. To address the mechanisms of maternal adaptation to hypoxia during implantation, we generated Hif2αd/d mice that conditionally lack HIF2α in the endometrium. In these mice, the blastocysts are closely apposed to the uterine epithelium but fail to sustain firm attachment to it, resulting in implantation failure. The uterine stromal cells of Hif2αd/d mice exhibited downregulation of metabolic factors, such as lactate and a subset of amino acids, and a distinct set of Rab GTPases that regulate secretion of extracellular vesicles (EVs), including microvesicles and exosomes. The EVs are known to mediate a novel mechanism of cell-cell communication. That hypoxia promotes EV secretion by certain cells support the concept that HIF2α-driven EV secretion by the endometrial stromal cells represents a critical adaptive response that promotes intercellular communication during implantation. In the proposed study, we will employ the Hif2αd/d mouse model to test the hypothesis that HIF2α regulates a novel paracrine signaling mechanism under the hypoxic conditions of early pregnancy by controlling EV trafficking in the maternal tissue and that it directs transfer of key signaling molecules and metabolites from the stromal to luminal epithelial cells to influence epithelial functionality during implantation. We have proposed three specific aims to test this hypothesis. In Aim 1, we will investigate HIF2α-mediated EV trafficking in endometrial stromal cells and identify the resident molecular cargo in EVs. In Aim 2, we will investigate the effects of EVs and metabolic factors secreted from endometrial stromal cells on the functionality of epithelial cells. In Aim 3, we will investigate the role of HIF2α in regulating the secretory function of human endometrial stromal cells. A multi-pronged approach, utilizing a combination of (i) a unique animal model harboring a specific defect in adaptive response (ii) functional analyses using mouse and human primary endometrial cells, and (iii) genomic and proteomic analyses to understand cell-to-cell trafficking mechanisms, will enable us to provide answers to important unresolved questions regarding the impact of hypoxia on stromal-epithelial communication controlling endometrial function during early pregnancy.

当囊胚附着在子宫腔上皮上时，着床就开始了，随后渗透到底层基质，牢固地嵌入子宫内膜。植入仍不清楚，新出现的证据表明，细胞间通讯子宫内膜上皮和基质细胞对于成功妊娠至关重要。我们最近的研究表明，调节这种串扰的信号仍然知之甚少。转录因子缺氧诱导因子 2 α (HIF2α) 在子宫内膜基质中选择性诱导植入时管腔上皮下的细胞这一发现很重要，因为它是长期以来，人们都知道着床期间的母体环境是缺氧的。着床期间母体适应缺氧的机制，我们生成了 Hif2αd/d 小鼠在这些小鼠的子宫内膜中条件性地缺乏 HIF2α，囊胚与子宫紧密相连。上皮，但无法维持其牢固附着，导致子宫基质细胞着床失败。 Hif2αd/d 小鼠表现出代谢因子的下调，例如乳酸和氨基酸的子集，并且一组独特的 Rab GTP 酶，可调节细胞外囊泡 (EV)（包括微泡）的分泌众所周知，EV 可以介导细胞间通讯的新机制。促进某些细胞分泌 EV 支持 HIF2α 驱动子宫内膜分泌 EV 的概念基质细胞代表了一种关键的适应性反应，可促进细胞间的通讯在拟议的研究中，我们将采用 Hif2αd/d 小鼠模型来检验以下假设： HIF2α 在妊娠早期缺氧条件下调节新型旁分泌信号机制控制母体组织中的 EV 运输并指导关键信号分子的转移从基质到管腔上皮细胞的代谢物在植入过程中影响上皮功能。我们提出了三个具体目标来检验这一假设。在目标 1 中，我们将研究 HIF2α 介导的。在目标 2 中，我们将研究子宫内膜基质细胞中的 EV 贩运并识别 EV 中的常驻分子货物。研究子宫内膜基质细胞分泌的 EV 和代谢因子对子宫内膜的影响在目标 3 中，我们将研究 HIF2α 在调节分泌中的作用。人类子宫内膜基质细胞的功能是一种多管齐下的方法，利用（i）独特的组合。具有适应性反应特定缺陷的动物模型（ii）使用小鼠和人类原代子宫内膜细胞，以及（iii）基因组和蛋白质组分析以了解细胞间的关系贩运机制，将使我们能够回答有关贩运人口的重要未决问题早期缺氧对控制子宫内膜功能的基质-上皮通讯的影响怀孕。