2012-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/659619摘要：滋養層母細胞的組織侵襲功能是否完全，影響胎盤的正常發育與否。甚至有些妊娠期病症與此亦息息相關，子癇前症就是一個例子。根據我們研究團隊研究結果顯示，Mucin 1及Mucin 15會隨著懷孕週數增加而提高在胎盤的表現量。而更重要的是Mucin 1在decidua的extravillous cytotrophoblasts (EVTs)的表現量亦隨著週數增加，而且在severe preeclampsia的EVTs，Mucin 1 positive rate更明顯增加。所以Mucins與胎盤發育及preeclampsia /IUGR的產生的關連性，自然不能忽略。我們知道像Mucins這類的glycoprotein，它們的生物功能表現，往往與它們的carbohydrate結構組成也有密切關聯。也就是說，在這些protein residues產生的glycosylation (post-translational modification)過程，也是影響mucin 的cell signaling功能的重要因素。而在protein scaffold 的serine或threonine residues加上N-acetyl- galactosamine 是mucin-type O-linked glycan biosynthesis的起始步驟，所以研究這個在glycosylation起始步驟的key enzyme — GalNac-transferase 2 (GALNT2)，將是接續前面研究很重要的焦點。所以我們設計了一個 3年期的計劃，依序研究(1)建立HTR-8/SV neo cells 的GALNT2 overexpression的stable clone，還有GALNT2 overexpression對EVTs 的cell signaling及表現invasive phenotype的影響。(2)以HUVEC為研究model，觀察GALNT2overexpression 的condition medium對於HUVEC的影響，尤其在VEGF, VEGFR-1, PlGF的表現及與integrin, Src的互動關係。(3) 建立GALNT2的Transgenic mice的HTR-8之 stable transfectants的Xenograft study，同時包括研究cellular signaling的pathway。我們這次提出的計畫將主要focus在GALNT2在胎盤發育過程中的表現，GALNT2對trophoblast cell invasion的影響，對於preeclampsia的相關性以及它們與mucin overexpression的功能表現的因果關連。我們未來可以經由利用它們的inhibitors帶入研究，將mucin 1 and/or GALNT2成為診斷的markers以及列入治療preeclampsia的therapeutic targets，其重要性不可忽視。此為前驅性研究。<br> Abstract: Trophoblast invasion is crucial for normal placenta development. The stringent regulation of trophoblast invasion is required for appropriate spiral artery remodeling in the maternal–fetal interface and essential for maintaining a normal pregnancy. Some adverse condition in pregnancy such as preeclampsia or IUGR had been associated with poor trophoblast invasion and disturbed remodeling.Our results demonstrated that MUC1 and MUC15 expression increased with gestational age of the human placenta. Furthermore, MUC1 overexpression significantly suppressed invasion of trophoblast-like JAR cells, which was associated with a decrease in MMP9 activity. Our recent data shows that MUC1 expression is elevated in severe preeclamptic placentas and suppresses trophoblast cell invasion via beta1-integrin signaling.Glycosylation is the common post-translational process in which saccharides are selectively added to specific protein residues in order to convey the cell signaling function. The enzyme polypeptide GalNAc-transferases 2 (GALNT2) plays a key role in mucin-type O-linked glycan biosynthesis by installing the initial GalNAc residue on the protein scaffold. Changes in glycans have been detected and implicated under various physiological and pathological conditions.Therefore we design a three year proposal to study the correlation between GALNT2 and IUGR/preeclampsia which may be mediated by active mucins (MUC1?). At first, we survey the biological effect of overexpression of GALNT2 to the trophoblast cell lines with or without MUC1 interactions. Second, we study the influence of the GALNT2 overexpression on vasomotor reaction in HUVEC culture. Third, we observe the phenotypic manifestation of pregnant GALNT2 transgenic mice and also changes after SiRNA knockdown.It will provide the knowledge how the trophoblast phenotypes are affected and regulated by surface mucin-like glycoproteins; and the gained information will allow us, in future studies, to develop new diagnostic and therapeutic reagents for preeclampsia and IUGR.This is a pioneer study.子癇前症滋養層母細胞組織侵襲醣化轉化酶臍靜脈內皮細胞preeclampsiatrophoblastinvasionglycosyltransferaseHUVEC