TLR3, TICAM1 and human rhinovirus infection

描述（由申请人提供）：TLR 3、TICAM 1和人鼻病毒感染人鼻病毒（RV）感染与普通感冒和哮喘的发展和/或恶化相关。迄今为止，我们和其他人已经报道了一个复杂的RV诱导的上皮促炎和抗病毒途径，其中涉及TLR 3，RIG 1，MDA 5和PKR。最近，我们发现TLR 3激活似乎是所有其他途径的上游，并且通过TICAM 1降解的TLR 3脱敏对于气道上皮的抗病毒防御至关重要。为了进一步阐明潜在的机制，我们建议首先在体外确定TLR 3脱敏对上皮抗病毒防御的影响。在这项研究中，我们将确定TICAM 1蛋白周转的动力学及其下游细胞的抗病毒反应诱导的dsRNA或RV。我们还将在此过程中确定细胞的抗病毒状态。最后，我们将过表达TICAM 1，以测试这是否可以恢复细胞的抗病毒反应。为了评估哮喘中TLR 3脱敏的潜在改变，我们将确定Th 2-细胞因子处理的上皮细胞是否具有改变的脱敏，以及与正常细胞相比，人哮喘上皮细胞在该过程中是否有缺陷。然后，我们建议确定TICAM 1降解的机制基础。在这个目标中，我们将测试一个新的假设，TICAM 1是通过分子伴侣介导的自噬降解。这是一个高风险和高影响的项目，非常适合R21机制。如果我们成功，我们将能够获得新的见解TLR 3-TICAM 1介导的气道抗病毒防御，并确定哮喘急性发作的危险因素。

DESCRIPTION (provided by applicant): TLR3, TICAM1 and human rhinovirus infection Human rhinovirus (RV) infection is associated with the common cold and asthma development and/or exacerbation. To date, we and others have reported a complex of RV-induced epithelial proinflammatory and antiviral pathways, which involve TLR3, RIG1, MDA5 and PKR. Recently, we have shown that TLR3 activation appears to be upstream of all other pathways, and TLR3 desensitization via TICAM1 degradation is critical to antiviral defense by airway epithelium. To further elucidate the underlying mechanism, we propose to first determine the impact of TLR3 desensitization on epithelial anti-viral defense in vitro. In this study, we will determine the kinetics of TICAM1 protein turnover and its downstream cellular antiviral responses induced by dsRNA or by RV. We will also determine the cellular antiviral status during this process. Lastly, we will overexpress TICAM1 to test if this could restore the cellular antiviral response. To evaluate the potential alteration of TLR3 desensitization in asthma, we will determine whether or not Th2-cytokine- treated epithelial cells have altered desensitization, and whether or not human asthmatic epithelial cells are defective in this process as compared to the normal cells. Then, we propose to determine the mechanistic basis of TICAM1 degradation. In this aim, we will test a novel hypothesis that TICAM1 is degraded via chaperone mediated autophagy. This is a high-risk and high-impact project that is well suited for R21 mechanism. If we succeed, we will be able to gain novel insight into the TLR3-TICAM1 mediated airway antiviral defense and to identify a risk factor for asthma exacerbation.