ALPHA-2 MACROGLOBULIN RECEPTOR, RAP, AND PREECLAMPSIA

ALPHA-2 巨球蛋白受体、RAP 和先兆子痫

• 批准号: 2227080
• 负责人: Dudley K. Strickland
• 金额: $ 22.24万
• 依托单位: AMERICAN NATIONAL RED CROSS
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-09-30 至 1998-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2227080
• 关键词: RNase protection assay; embryo implantation; fibrinolysis; human tissue; immunocytochemistry; immunoprecipitation; ligands; low density lipoprotein; macroglobulins; membrane proteins; messenger RNA; molecular pathology; northern blottings; placenta; plasminogen activator; preeclampsia; receptor; receptor binding; receptor expression; surface property; tissue inhibitor of metalloproteinases; trophoblast; western blottings

Normal pregnancy is accompanied by extensive changes in the blood coagulation and fibrinolytic system. Preeclampsia is a moor obstetrical problem characterized by hypertension, proteinuria, increased activation of blood coagulation, and decreased activity of the fibrinolytic pathway. A feature of this disorder is the shallow invasion of trophoblast cells into the uterus. The processes of normal embryo implantation and placentation are dependent upon the regulated expression of proteolytic enzymes that degrade the uterine matrix. There is substantial evidence indicating that the alpha2-macroglobulin receptor (alpha2MR) plays an essential role in mediating trophoblast invasive activity, possibly by facilitating the removal of urokinase type plasminogen activator (u-PA) complexed to plasminogen inhibitor complex (PAI-1) from cell surface receptors. We propose that trophoblast invasion requires a constant turnover of inhibited proteinases on the cell surface. We also hypothesize that expression or function of the alpha2MR is abnormal in pregnancies complicated by preeclampsia where trophoblast invasion is shallow. The goals of the proposed research project are to explore the role of the alpha2MR and the 39 kDa receptor-associated protein (RAP) in the implantation process. The first specific aim is to define the role of alpha2MR in removing uPA:PAI-1 complexes from the cell surface, and to determine if the alpha2MR mediates catabolism of metalloproteinase complexed to their inhibitor, tissue inhibitor of metalloproteinases (TIMP). The second specific alm is designed to explore the role of RAP in trophoblast invasion. This molecule has been observed to block binding of all ligands to alpha2MR. RAP function will be examined by using established assays which quantitates cell movement through a layer of Matrigel on a porous membrane and assays that measure the extracellular matrix degradation by cells from mouse blastocyst outgrowths. The third specific alm is to define the expression of the alpha2MR in human placenta at different gestational ages and in placental bed biopsies. The receptor and RAP will be localized with specific antibodies to determine the spatial and temporal patterns of expression. The expression of alpha2MR and RAP in placental tissue and placental bed biopsies of pregnancies complicated by preeclampsia will also be examined. Immunocytochemistry, Western blotting and mRNA analyses will be employed. The proposed studies will provide a comprehensive description of the alpha2MR and its endogenous ligand, RAP, in trophoblast invasion, and will test the hypothesis that abnormal receptor expression or function is associated with preeclampsia. If validated, we will have contributed a model for the molecular basis of this important disorder.

正常怀孕伴随着血液的广泛变化 凝血和纤维蛋白水解系统。 子痫前期是摩尔的产科 以高血压，蛋白尿，激活增加为特征的问题 血液凝结，纤维蛋白水解途径的活性降低。 该疾病的一个特征是滋养细胞的浅侵袭 进入子宫。 正常胚胎植入和 胎盘取决于蛋白水解的调节表达 降解子宫基质的酶。有大量证据 表明Alpha2-macrogobolin受体（alpha2mr）播放 可能由 促进去除尿激酶型纤溶酶原激活剂（U-PA） 从细胞表面复合到纤溶酶原抑制剂复合物（PAI-1） 受体。我们建议滋养细胞入侵需要常数 细胞表面抑制蛋白酶的周转率。我们也假设 α2MR的表达或功能在怀孕中是异常的 因滋养细胞侵袭浅的先兆子痫而复杂。这 拟议的研究项目的目标是探索 α2MR和39 kDa受体相关蛋白（RAP） 植入过程。第一个具体目的是定义 alpha2mr删除UPA：从细胞表面和到 确定α2MR是否介导金属蛋白酶的分解代谢 将金属蛋白酶的组织抑制剂复合到其抑制剂 （TIMP）。 第二个特定ALM旨在探索说唱在 滋养细胞入侵。 已经观察到该分子阻止 所有配体到α2MR。 RAP功能将通过使用 已建立的测定法，该测定法可以通过一层量化细胞运动 在多孔膜上的矩阵和测量细胞外的测定 小鼠胚泡产物的细胞降解基质。 第三 特定的ALM是定义人胎盘中α2MR的表达 在不同的胎龄和胎盘床活检中。受体 RAP将以特定的抗体定位，以确定 表达的空间和时间模式。 α2MR的表达 和胎盘组织和胎盘床活检的说唱 还将检查复杂的先兆子痫。免疫细胞化学， 将采用蛋白质印迹和mRNA分析。 提出的研究 将提供对alpha2Mr及其α2MR的全面描述 内源配体，说唱，在滋养细胞入侵中，将测试 假设异常受体表达或功能是相关的 与先兆子痫。 如果得到验证，我们将为 这种重要疾病的分子基础。