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

描述(申请人提供)：到目前为止，我们的工作重点是了解摄取的哺乳动物转化生长因子β1是如何被激活的，调控蚊子Smad信号和内源性转化生长因子βS，并最终减少斯氏疟原虫中的疟疾寄生虫载量。这项工作导致了在哺乳动物免疫和炎症领域的进展指导下，对发生这种物种间串扰的信号通路的研究。在哺乳动物中，有四条相互作用的调节通路与免疫相关：核因子-βB通路和三条丝裂原激活蛋白激酶(MAPK)通路，包括JNK、ERK和p38依赖通路。我们已经证明，斯氏艾美耳球虫细胞中的转化生长因子-1信号受氧化还原化学调节，涉及斯氏艾美耳球虫ERK、JNK和p38同源物的差异激活。我们还确定了其他血粉来源的因子，包括胰岛素、两种寄生虫毒素和两种哺乳动物炎症介质，它们与转化生长因子-1一样，可能作为斯氏疟原虫细胞的信号。事实上，我们的初步数据表明，这些因素也调节斯氏杆菌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--包括蚊子宿主、哺乳动物宿主和寄生虫--作为一个整体来阻止疟疾感染。我们假设，一个协调的通路网络(MAPK、Smads、NF-βB)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，必须由生态系统的所有三个成员解释。考虑到单一物种炎症反应的复杂性，这一挑战令人望而生畏。然而，我们已经证明，这种复杂性可以通过相互关联的实验方法和计算模拟来理性地解决。因此，我们将利用我们在这一系统上的经验，以及在多个尺度上对炎症的计算模拟，从细胞内水平到多生物水平，包括疟疾背景下物种间串扰的初步模型。我们认为，这种综合的方法将使我们不仅能够识别在免疫串扰过程中操作的机制，而且还可以解释系统中的意外行为，并定义对斯氏疟原虫寄生虫发展具有最大潜在影响的“主开关”-串扰细胞外因子和信号通路组件。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做出反应并将其消灭的单个基因产物上，但很少或根本没有关于这些反应的协调调控的信息。我们的研究将阐明这种协调，并开发蚊子、寄生虫和哺乳动物宿主的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展的“总开关”。从长远来看，我们认为这些信息将有助于新的疟疾控制方法。

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.