Placental Gene Expression Profile in Preeclampsia

描述(由申请人提供)： 本申请是为响应PAR-01-066《KO8获奖者小额资助计划》而提交的，并与NIH KO8 DK02825-03相关联。在KO8基金的支持下，申请人完成了博士后研究培训，开始了独立的研究生涯。这笔赠款将增加财政独立性，并为申请者提供在一个相对较新的研究领域产生数据的机会，这些数据可以构成RO1申请的基础。子痫前期(PE)是一种以严重高血压、蛋白尿和水肿为特征的疾病，发生率为5%。内皮功能障碍在本病的发病机制中起着重要作用，但其病因和机制尚不清楚。有越来越多的证据表明PE的致病模型是低氧胎盘单位分泌一种因子进入母体循环，导致全身血管内皮细胞功能障碍。在提案的前半部分，申请人希望使用正常和先兆子痫胎盘的mRNA表达谱来识别PE中的从头靶点。申请人希望使用生物信息学工具，如等级聚类和k-均值来识别将预测PE的发生和严重程度的基因簇。这些研究将导致识别生物标记物，这可能用于诊断，并可能导致这种复杂疾病的新疗法。在使用先兆子痫胎盘基因表达谱的初步实验中，申请人已经确定“sFlt-1”(可溶性FMS样酪氨酸激酶)是PE患者上调的一个因素。Flt-1是血管内皮生长因子(VEGF)和胎盘生长因子(PGF)的酪氨酸激酶受体。SFlt-1是FIT-1的一种分泌型剪接变异体(缺少跨膜区和胞浆区)，通过阻止VEGF和PGF与其细胞表面受体相互作用，是一种强有力的血管内皮生长因子和前列腺素F的拮抗剂。该提案的后半部分将定义sFlt-1在PE发病机制中的作用。我们将检验这样一种假设，即由先兆子痫胎盘释放的sFlt-1与部分或全部与PE相关的体外和体内表型有关。这些重点研究形成了一个理解PE发病机制的框架。申请人于2001年7月刚开始其独立调查员的职业生涯。申请者积极进取，完全致力于作为一名内科科学家的职业生涯。申请者在内皮细胞生物学方面的经验、不同合作者的专业知识以及哈佛医学院的支持性环境为申请者提供了一个理想的论坛，不仅可以实现这项建议的目标，还可以成为一名成功的独立研究人员。

DESCRIPTION (provided by applicant): This application for RO3 award is submitted in response to PAR-01-066, " Small grant program for KO8 recipients" and is linked to NIH KO8 DK02825-03. With the support of the KO8 funding, the applicant has completed his post-doctoral research training and has begun an independent research career. This grant will increase fiscal independence and provide an opportunity to generate data in a relatively new area of research for the applicant that can form the basis for an RO1 application. Preeclampsia (PE) is a disease characterized by severe hypertension, proteinuria and edema and occurs in 5% of all pregnancies. Endothelial dysfunction plays an important role in the pathogenesis of this disorder; however the etiology and mechanisms are still unknown. There is accumulating evidence for a pathogenic model for PE whereby a hypoxic feto-placental unit secretes a factor into the maternal circulation causing generalized endothelial cell dysfunction. In the first half of the proposal the applicant wishes to identify de novo targets in PE using mRNA expression profiling in normal and preeclamptic placentas. The applicant wishes to use bioinformatic tools such as hierarchical clustering and k-means to identify clusters of genes that will predict the occurrence and severity of PE. These studies will lead to identification of biomarkers, which may be used diagnostically and potentially lead to new therapies for this complex disorder. In preliminary experiments using gene expression profile from preeclamptic placentas, the applicant has identified "sflt-1" (soluble fms-like tyrosine kinase) as a factor that is up-regulated in patients with PE. Flt-1 is one of the tyrosine kinase receptors for vascular endothelial growth factor (VEGF) and placental growth factor (PGF). Sflt-1, a secreted splice variant of fit-1 (lacking the transmembrane and cytoplasmic domains) is a potent antagonist of VEGF and PGF, by preventing both VEGF and PGF from interacting with its cell-surface receptor. The second half of the proposal will be to define the role of sflt-1 in the pathogenesis of PE. We will test the hypothesis that sflt-1, which is released by preeclamptic placenta, is responsible for some or all of the in vitro and in vivo phenotypes associated with PE. These focused studies form the beginnings of a framework to understand the pathogenesis of PE. The applicant has just begun his career as an independent investigator in July 2001. The applicant is highly motivated and absolutely committed to a career as a physician-scientist. The experience of the applicant in endothelial cell biology, the expertise of various collaborators and the supportive environment at Harvard Medical School provides an ideal forum for the applicant to not only realize the objectives of this proposal but also to become a successful independent investigator.