基于磷酸化蛋白质组学发现的血吸虫重要信号分子调控虫体生殖发育的分子机理研究

项目名称： 基于磷酸化蛋白质组学发现的血吸虫重要信号分子调控虫体生殖发育的分子机理研究

项目编号： No.31472187

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 水产学、兽医学

项目作者： 程国锋

作者单位： 中国农业科学院上海兽医研究所

项目金额： 83万元

中文摘要： 血吸虫雌雄合抱是雌虫性发育成熟的关键，性发育成熟的雌虫所产大量虫卵又是造成宿主病理损害和疾病传播的根本原因。目前，有关血吸虫生殖发育的分子机理认识仍非常有限。基于日本血吸虫磷酸化蛋白质组研究构建的蛋白质相互作用图，发现了数个重要调控节点和对话点(HSP90，GSK3Beta 和p38MAPK)。我们已研究表明阻断血吸虫HSP90 可显著降低动物体内虫体的寄生及肝脏的虫卵量，提示关键信号分子在血吸虫生殖发育和产卵中的重要作用。为此，本项目拟利用免疫共沉淀、酵母双杂交和RNAi 等技术研究重要调控节点HSP90，GSK3Beta 和p38MAPK 的分子功能及其相互作用分子，发现其参与血吸虫发育生殖和产卵的信号通路，探索干扰关键分子以阻断虫体发育成熟和产卵的可行性。研究结果不仅对阐明血吸虫的生殖发育机理具有重要的理论意义，还可为研发抗血吸虫生殖发育疫苗和药物提供新思路，具有重要的现实意义。

中文关键词： 日本血吸虫；分子生物学；信号通路；分子机制；生殖发育

英文摘要： Schistosomes are dioecious trematodes responsible for schistosomiasis in humans and other animals. Sexual maturation and subsequent egg production in female schistosomes require a permanent pairing with male schistosomes at the gynecophornal canal. Matured female schistosomes produce a large number of eggs which are primarily responsible for the pathogenesis of schistosomiasis as well as dissemination of the parasite. However, the current understanding of molecular mechanism of female worm maturation induced by the male worm after pairing is very limited. In our previous study, a phosphoproteomic approach was employed to investigate the phosphoproteins in Schistosoma japonicum. We then built a protein-protein interaction map (PPI) based on several hundred identified phosphoproteins in Schistosome japonicum by using comparative genomics combined with bioinformatics. The resulted PPI indicated that there were several potentially important regulatory nodes and crosstalk points mainly including Heat shock protein 90 (HSP90), Glycogen synthase kinase 3 beta (GSK3B), and p38 motigen-activated protein kinase (p38MAPK). Our preliminary study showed that the block of HSP90 function in Schistosoma japonicum by applying drug inhibitor led to the significant reduction of worms in the final host and the eggs in the host's liver, suggesting these molecules in the regulatory nodes and crosstalk points of PPI play an important role in the regulation of schistosome development, sexual maturation and egg production. Additionally, recent study indicated that HSP90 could facilitate/mature a large number of signaling molecules involved in many biological processes such as GSK3B and p38MAPK in both human and yeast. Here, we propose to use co-immunoprecipitation, yeast two hybrid system and RNA interference (RNAi) to experimentally determine the functions of the HSP90, GSK3B and p38MAPK in female schistosomes for sexual maturation and egg production and also investigate their partners and signal pathways involved in schistosome development and sexual maturation. Then, we plan to apply RNAi and chemical inhibitors to block the functions of these molecules and/or their involved signal pathways to evaluate the potential of these molecules as drug targets against schistosomiasis. The expected results of the proposal will not only provide important information for theoretically understanding the mechanism of schistosome development and sexual maturation but also may facilitate to develop effective anti-schistosome vaccines and drugs for schistosomiasis control.

英文关键词： Schistosoma japonicum;Molecular Biology;Signal tranduction;molecular mechanisms;Sexual maturation and development