Cell-cell communication mediated by fluid flows

项目摘要 细胞可以检测到穿过其表面的外部流体流动。这种流动，以及它们诱导的流动信号事件， 对器官发育、体内平衡、癌症传播和宿主-微生物相互作用至关重要，但我们 我们对细胞如何感知和响应它们的水动力环境知之甚少。该研究计划旨在 揭示细胞通过液体流动进行交流的基本机制。具体来说，我们调查 细胞如何产生和感知液体流动，以及感觉细胞如何对流动产生的信号做出反应。完成网站 关于流信号的新的基础知识，我们使用完善的和高度易处理的左-右 斑马鱼胚胎的图案系统。在这个系统中，这是服从遗传扰动，以及 在高分辨率和定量成像中，流动信号抑制了关键靶基因dand 5的表达， 感觉细胞在我们之前的工作中，我们发现Pkd 1 l1，一种大的多囊蛋白膜蛋白，对于 流动诱导的Dand 5抑制。在我们提出的工作的一个方面，我们将测试我们的假设，Pkd 1 l1是 通过确定细胞类型以及亚细胞作用位点的流动信号感觉组件，其中 pkd 1 l1在流量信号下游起作用。我们还研究了机械敏感的功能作用， Pkd 1 l1内的结构域在流量调节信号传导中。在第二个项目中，我们解决了 了解流信号如何在感觉细胞内转导。这项工作将使用我们的优化 CRISPR方法可以发现和验证新的流动信号转导机制，这将 显着扩大了我们对感觉事件下游作用机制的理解， 细胞内信号。在第三个项目中，我们将使用已知的dand 5转录后抑制， 具体地发生在流量信号的左侧下游，作为用于生成以下新知识的模型： 感觉细胞如何对血流信号做出反应这包括确定3 '非翻译区的作用， RNA结合蛋白和microRNA如何在流动信号下游起作用以抑制基因表达。这 我们正在开发的定量流动信号传导途径输出的新型体内报告物将有助于这项工作。 总的来说，这项研究将揭示细胞通过流动信号进行通信的新机制。而 化学信号转导级联（Hedgehog、Wnt等）被广泛研究，流量信号通路 代表了一个新的领域，有许多基本原理等待被发现。我们的工作利用了 斑马鱼易于处理的左右图案系统将揭示新的流动信号感知原理， 转导，以及细胞如何响应流动信号。由于异常血流信号发生在许多疾病中， 这将为改善人类健康提供新的机会。

Project Summary Cells can detect external fluid flows across their surface. Such flows, and the flow signaling events they induce, are critical for organ development, homeostasis, cancer dissemination, and host-microbe interactions, yet we know little about how cells sense and respond to their hydrodynamic environment. This research program aims to reveal fundamental mechanisms by which cells communicate through fluid flows. Specifically, we investigate how cells generate and sense fluid flows, and how sensory cells respond to flow-derived signals. To generate new fundamental knowledge about flow signaling, we use the well-established and highly tractable left-right patterning system of zebrafish embryos. In this system, which is amenable to genetic perturbation as well as high-resolution and quantitative imaging, a flow signal represses the expression of a key target gene, dand5, in sensory cells. In our prior work, we discovered that Pkd1l1, a large Polycystin membrane protein, is critical for flow-induced dand5 repression. In one aspect of our proposed work, we will test our hypothesis that Pkd1l1 is a flow signal sensory component by determining the cell types, as well as sub-cellular site of action, in which Pkd1l1 functions downstream of flow signals. We also investigate the functional role of mechanosensitive domains within Pkd1l1 in flow-regulated signaling. In a second project, we address the major gap in understanding of how flow signals are transduced within sensory cells. This work will use our optimized CRISPR approaches to discover and validate new flow signal transduction machinery, something which will markedly expand our understanding of the mechanisms acting downstream of sensory events to transduce signals intracellularly. In a third project, we will use the known post-transcriptional repression of dand5 that occurs specifically on the left side downstream of the flow signal as a model for generating new knowledge of how sensory cells respond to flow signals. This includes determining the role of the 3’untranslated region and how RNA-binding proteins and microRNAs act downstream of flow signals to repress gene expression. This work will be aided by novel in vivo reporters we are developing which quantify flow signaling pathway outputs. Overall, this research will uncover novel mechanisms by which cells communicate through flow signals. While chemical signal transduction cascades (Hedgehog, Wnt, etc.) are widely studied, flow signaling pathways represent a new frontier, with many fundamental principles waiting to be discovered. Our work using the highly tractable left-right patterning system of zebrafish will reveal new principles of flow signal sensation and transduction, as well as how cells respond to flow signals. Since aberrant flow signals occur in many disease states, this will open new opportunities to improve human health.