Regulation of maspin in placental development

描述(申请人提供)：胎盘侵入是一个高度调控的过程，损害会导致病理产科疾病，如先兆子痫和宫内生长受限(IUGR)。进一步鉴定影响细胞滋养层细胞侵袭能力的因素将有助于更好地理解这一独特的发育过程。我们已经证明了胎盘侵袭和发育中新的肿瘤抑制基因maspin的表达有一个特定的时间线。我们的发现表明，Maspin在妊娠晚期表达最多，在妊娠早期表达水平较低。这项建议的具体目的是：1)检测maspin上调在体外对人细胞滋养层细胞侵袭、运动和形态发生的生物学影响(S)；2)研究低氧在调节细胞滋养层细胞maspin表达中的作用3)比较有先兆子痫和无先兆子痫的足月和早产孕妇胎盘中maspin/锰超氧化物歧化酶基因和蛋白的丰度。研究设计将包括在第一次和第二次试验中使用腺病毒方法将maspin转染到原代滋养层细胞培养中，并检测这种转染对细胞侵袭、迁移、运动和形态发生的影响。接下来，我们将研究低氧对maspin表达的影响，以及低氧诱导因子在介导这些影响中的作用。最后，将使用缺氧和受损的细胞滋养细胞侵袭的体内模型来证实我们的假设，即先兆子痫。Maspin在子痫前期患者胎盘和正常对照胎盘中的表达将通过实时定量聚合酶链式反应进行比较。该项目的长期目标有两个，第一，了解Maspin等肿瘤抑制基因在人类胎盘发育中的意义和作用机制(S)；第二，确定由Maspin表达变化引起的妊娠病理过程。这项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的治疗策略有深远的影响，如先兆子痫和宫内发育迟缓。此外，任何可能获得的关于maspin在胚胎发育过程中可能扮演的角色的重要新信息，都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.