TRPC6 Regulation of Lung Endothelial Barrier Function

在项目3中提出的研究将调查的基本问题，即Ca 2+进入介导的瞬时 受体电位阳离子通道6（TRPC 6）是一种受体操纵的通道（ROC），是一种关键的决定因素， 增加内皮通透性。我们将检验TRPC 6是细胞内流的关键途径这一假设。 触发内皮收缩机制RhoA和MLCK内皮亚型激活的Ca 2 + （EC-MLCK）。如果这一假设是正确的，它将证明TRPC 6和蛋白质之间的基本联系。 RhoA-EC-MLCK通路。我们还假设，根据我们的支持数据， TRPC 6激活的信号传导上调TRPC 1（一种存储操作的通道）活性的关系 （SOC）。因此，我们的概念是TRPC 6通过激活RhoA和EC-MLCK两者来控制细胞的状态。 此外，TRPC 6激活上调TRPC 1活性的信号，从而 延长和/或放大内皮渗透性反应。我们的具体目标是确定：1） 内皮ROC，TRPC 6，与SOC，TRPC 1相互作用，在介导Ca 2+信号依赖性 肺血管通透性增加; 2）TRPC 6激活的信号转导在介导RhoA和ECMLCK中的作用 激活和这些效应物在介导肺血管内皮细胞增加中的需要 3）TRPC 6下游的EC-MLCK在上调TRPC 1活性中的作用，及其 增加肺内皮通透性的后果。这些研究将利用 内皮细胞和肺，这些细胞来自携带特定基因缺失的小鼠。我们相信这些研究将 为Ca 2+介导的肺血管通透性增加和水肿提供了新的认识 阵这些新的发现将有助于揭示TRPC 6和TRPC之间的相互作用的细节。 TRPC 1信号肺内皮通透性的Ca 2+依赖性增加。这些研究预计将 提供了抑制TRPC 6和TRPC 1活性的新的治疗策略，从而消除或逆转 肺血管渗漏

Studies proposed in Project 3 will investigate the fundamental question that Ca2+ entry mediated by transient receptor potential cation channel 6 (TRPC6), a receptor-operated channel (ROC), is a critical determinant of increased endothelial permeability. We will test the hypothesis that TRPC6 is the critical influx pathway for Ca2+ that triggers the activation of endothelial contractile mechanisms RhoA and endothelial isoform of MLCK (EC-MLCK). If this hypothesis is correct, it would demonstrate a fundamental link between TRPC6 and the RhoA-EC-MLCK pathway. We also postulate based on our Supporting Data a potentially important relationship by which TRPC6-activated signaling upregulates the activity of TRPC1, a store-operated channel (SOC). Thus, our concept is that TRPC6 by activating both RhoA and EC-MLCK controls the state of endothelial contraction and additionally that TRPC6 activates signals that upregulate TRPC1 activity, thereby protracting and/or amplifying the endothelial permeability response. Our Specific Aims are to determine: 1) the role of the endothelial ROC, TRPC6, interaction with the SOC, TRPC1, in mediating Ca2+ signaling-dependent increase in lung vascular permeability; 2) the role of TRPC6-activated signaling in mediafing RhoA and ECMLCK activation and the requirement of these effectors in mediating increased lung vascular endothelial permeability and 3) the role of EC-MLCK downstream of TRPC6 in upregulating TRPC1 activity, and its consequences in increasing lung endothelial permeability. These studies will be carried out utilizing endothelial cells and lungs from mice carrying deletions of specific genes. We believe that the studies will provide a new understanding of the Ca2+-mediated increase in lung vascular permeability and edema formation. These new findings will help to unravel the details of how the interactions between TRPC6 and TRPC1 signals the Ca2+-dependent increase in lung endothelial permeability. The studies are expected to offer a novel therapeutic strategy for inhibiting TRPC6 and TRPC1 activities, thereby abrogating or reversing lung vascular leakiness.