Regulation of maspin in placental development

描述（由申请人提供）：胎盘侵犯是一个高度调控的过程，损害导致病理性产科状况，如先兆子痫和子宫内生长受限（IUGR）。进一步确定在改变细胞滋养细胞侵袭能力中起作用的因素将有助于更好地理解这一独特的发育过程。我们已经证明了maspin，一种新的肿瘤抑制基因在胎盘侵袭和发育中的具体表达时间。我们的研究结果表明，maspin在妊娠晚期表达最多，而在妊娠早期表达水平较低。本研究的具体目的是：1)测定maspin在人细胞滋养层细胞中上调对体外侵袭性、运动性和形态发生的生物学影响；2)研究缺氧对maspin在细胞滋养层细胞中表达的调节作用；3)比较足月和早产妇女胎盘中maspin/锰超氧化物歧化酶mRNA和蛋白的丰度，伴和不伴先兆子痫。研究设计将包括在第一次和第二次测试中使用腺病毒方法将maspin转染到原代滋养细胞培养中，并检查这种转染对细胞侵袭、迁移、运动和形态发生的影响。接下来，我们将探讨缺氧对maspin表达的影响以及缺氧诱导因子在这些影响中的作用。最后，我们将使用缺氧和细胞滋养细胞侵袭受损的体内模型，即子痫前期，来证实我们的假设。Maspin在子痫前期妇女胎盘和正常对照胎盘中的表达将采用实时PCR进行比较。该项目的长期目标有两个方面：一是了解肿瘤抑制基因如maspin在人类胎盘发育中的作用及其意义；二是确定maspin表达改变引起的妊娠病理过程。本研究揭示的信息可能对与细胞滋养细胞侵袭减少相关的临床疾病（如子痫前期和IUGR）的治疗策略具有深远的意义。此外，可能获得的关于maspin在胚胎发育过程中假定作用的任何重要新信息可能有助于阐明其在肿瘤进展过程中丢失的生物学意义。

DESCRIPTION (provided by applicant): Placental invasion is a highly regulated process and impairment results in pathologic obstetric conditions such as preeclampsia and intra-uterine growth restriction (IUGR). Further identification of factors that play a role in modifying this invasive ability of cytotrophoblasts will contribute to a better understanding of this unique developmental process. We have demonstrated a specific timeline for the expression of maspin, novel tumor suppressor gene in placental invasion and development. Our findings show that maspin is maximally expressed in the third trimester with low levels of expression in the first trimester of pregnancy. The specific aims of this proposal are 1) To measure the biological consequence(s) of maspin up regulation in human cytotrophoblasts with respect to invasion, motility and morphogenesis in vitro 2) Examine the role of hypoxia in regulating maspin expression in cytotrophoblasts 3) Compare the abundance of maspin/manganese superoxide dismutase mRNA and protein in placentas from women with term and preterm deliveries, with and without preeclampsia. The research design will include using adenoviral approach to transfect maspin into primary trophoblast cultures during the first and second testers and examine the effects of this transfection on cell invasion, migration, motility and morphogenesis. Next, the effects of hypoxia on maspin expression and the role of hypoxia-inducible factor in mediating these effects will be examined. Finally, will confirm our hypothesis using an in vivo model of hypoxia and impaired cytotrophoblast invasion namely preeclampsia. Maspin expression will be compared in placentae from women with preeclampsia to those from normal controls using real-time PCR. The long-term objectives of this project are two-fold, first, to understand the significance and mechanism(s) of action of tumor suppressor genes such as maspin in human placental development second, to identify pathological processes during gestation arising from alterations of maspin expression. Information revealed in this study could have profound implications on therapeutic strategies for clinical conditions associated with decreased cytotrophoblast invasion such as preeclampsia and IUGR. In addition, any important new information that may be obtained about the putative role of maspin during embryonic development may help elucidate the biological significance of its loss during tumor progression.