Systems biology approach to MAPK regulation of malaria infection in A. stephensi

描述（由申请人提供）：迄今为止，我们的工作主要集中在了解摄入的哺乳动物转化生长因子(TGF)-¿1如何被激活，调节蚊子的Smad信号和内源性TGF-¿s，并最终减少斯氏伊蚊的疟原虫负荷。在哺乳动物免疫和炎症领域的进展指导下，这项工作导致了对这种物种间串扰发生的信号通路的研究。在哺乳动物中，有四种相互作用的调节途径与免疫有关：核因子(NF)-？B途径和三种丝裂原活化蛋白激酶（MAPK）途径，包括JNK、ERK和p38依赖性途径。我们已经证明史蒂芬氏霉细胞中的TGF-¿1信号是由氧化还原化学调节的，并涉及史蒂芬氏霉的ERK、JNK和p38同源物的差异激活。我们还发现了其他血源性因子，包括胰岛素、两种寄生虫毒素和两种哺乳动物炎症介质，如TGF-¿1，可能作为斯蒂芬假体细胞的信号。事实上，我们的初步数据表明，这些因子也调节斯蒂芬氏霉ERK、JNK和p38的激活。我们的长期目标是操纵一个高度复杂的生态系统——它由蚊子宿主、哺乳动物宿主和寄生虫组成——作为一个整体，以阻止疟疾感染。我们假设一个协调的通路网络(MAPKs, Smads, NF-？B)调节蚊子对感染的反应。此外，由这些信号通路驱动的炎症结果启动了新的信号，这些信号必须由这个生态系统的所有三个成员来解释。考虑到单一物种的炎症反应的复杂性，这一挑战是艰巨的。然而，我们已经表明，这种复杂性可以通过相互关联的实验方法和计算模拟来合理地解决。因此，我们将利用我们在这个系统上的经验，以及在从细胞内水平到多有机体水平的多个尺度上对炎症进行计算模拟，包括疟疾环境下物种间串扰的初步模型。我们提出，这种综合方法将使我们不仅能够辨别在免疫串扰过程中运作的机制，而且还可以解释系统中的意外行为，并定义“主开关”-串扰细胞外因子和信号通路成分-对斯蒂芬氏拟虫的寄生虫发育具有最大的潜在影响。公共卫生相关性：斯氏按蚊是人类疟疾寄生虫恶性疟原虫的重要媒介。许多研究都集中在对这些寄生虫作出反应和消灭的单个基因产物上，但关于这些反应的协调调节的信息很少或没有。我们的研究将阐明这种协调，并建立蚊子、寄生虫和哺乳动物宿主生物界面的数学和统计模型。这些研究将有助于确定控制蚊子体内寄生虫发育的“主开关”。从长远来看，我们认为这些信息将有助于开发新的疟疾控制方法。

DESCRIPTION (provided by applicant): Our work to date has focused on understanding how ingested mammalian transforming growth factor (TGF)-¿1 is activated, regulates mosquito Smad signaling and endogenous TGF-¿s, and ultimately reduces malaria parasite loads in A. stephensi. This work led to the study of signaling pathways by which this inter-species crosstalk occurs, guided by advances in the field of mammalian immunity and inflammation. In mammals, four interacting regulatory pathways are associated with immunity: the nuclear factor (NF)-?B pathway and the three mitogen-activated protein kinase (MAPK) pathways, including JNK, ERK and p38-dependent pathways. We have shown that TGF-¿1 signaling in A. stephensi cells is regulated by redox chemistry and involves differential activation of the A. stephensi homologs of ERK, JNK, and p38. We have also identified other bloodmeal-derived factors including insulin, two parasite toxins, and two mammalian inflammatory mediators that, like TGF-¿1, may function as signals to A. stephensi cells. Indeed, our preliminary data suggest that these factors also regulate the activation of A. stephensi ERK, JNK, and p38. Our long-term goal is to manipulate a highly complex ecological system--which consists of the mosquito host, the mammalian host, and the parasite--as a whole in order to block malaria infection. We hypothesize that a coordinated network of pathways (MAPKs, Smads, NF-?B) regulates the mosquito response to infection. Moreover, the inflammatory outcomes driven by these signaling pathways set in motion new signals that must be interpreted by all three members of this ecosystem. This challenge is daunting, given the complexity of the inflammatory response in a single species. Yet, we have shown that this complexity can be addressed rationally through inter-connected experimental approaches and computational simulations. As such, we will leverage our experience with this system and with computational simulations of inflammation at multiple scales, from the intracellular level to the multi-organismal level, including preliminary models of inter-species crosstalk in the setting of malaria. We propose that this integrated approach will allow us to discern not only the mechanisms operant in the process of immune crosstalk, but also explain unexpected behavior in the system and define the "master switches" - the crosstalking extracellular factors and signaling pathway components - that have the greatest potential impact on parasite development in A. stephensi. PUBLIC HEALTH RELEVANCE: The mosquito Anopheles stephensi is an important vector of the human malaria parasite Plasmodium falciparum. Many studies have focused on individual gene products that respond to and destroy these parasites, but there is little to no information on the coordinated regulation of these responses. Our studies will elucidate this coordination and develop mathematical and statistical models of the biological interface of the mosquito, the parasite, and the mammalian host. These studies will serve to identify the "master switches" that control parasite development in the mosquito. In the long-term, we propose that this information will contribute to novel malaria control methods.