Prevention of adenovirus pathogenesis through downregulation of the apical adenovirus receptor

项目总结/摘要 腺病毒（AdenovirusAdV）是常见的人类病原体，在健康个体中引起典型的感冒症状。 急性呼吸窘迫综合征（ARDS），死亡率高达50%， 特别是在高度易感、免疫抑制的人群中，新的、潜在致命的变异继续存在， 每年都会出现。没有专门预防或治疗AdV感染的治疗剂存在， 新的治疗方法将预防与AdV感染相关的发病率和潜在死亡率。 本提案的目的是确定的作用机制和抗腺病毒疗效的 降低顶端柯萨奇病毒和腺病毒受体（CARex 8）蛋白表达的新分子。 我们的中心假设是阻断MAGI-1和CARex 8之间相互作用的诱饵肽， 在气道中的根尖CARex 8蛋白以消除AdV进入和发病机制。我们将测试我们的假设 使用具有Dox诱导型CAREx8表达的极化模型上皮， 啮齿类动物气道上皮细胞来验证我们的发现，棉鼠模型来评估野生型腺病毒的发病机制 在免疫活性动物和环磷酰胺免疫抑制动物中，有两个特定目的： 目的1：探讨MAGI-1激活肽和腺病毒介导的蛋白降解机制。 CAREx8的版本。完成这一目标将允许识别和开发新颖且多的 需要的目标和方法，以防止感染和传播的致病性野生型腺病毒， B组柯萨奇病毒。 目的2：确定MAGI-1 PDZ 1结合肽对野生型腺病毒的保护作用。 Rus感染棉鼠。这一目标的实现将提供原理证明，即AdV感染可以 通过减少其主要受体来阻止，并可能挽救严重感染或免疫抑制的个体的生命。 活力四射。 总体影响：了解调节表达和定位的机制， CARex 8，以及这种独特的亚型如何介导病毒进入极化上皮的顶端表面是关键的。 呼吁了解病毒的传播，组织嗜性和发病机制。圆满完成了亲- 提出的目标不仅将确定调控顶端腺病毒表达的细胞机制， 受体，但也调节病毒结合和感染的机制。这将使THERA的可行性- 降低气道上皮对腺病毒感染和致病性的易感性的抗菌剂 在免疫活性和免疫抑制条件下的感染前和活动性感染期间。我们 最终，我们希望这些目标能够导致新的抗病毒干预措施，这些措施也可能阻止其他病毒 它使用CAR作为主要受体，并提供了对其他相关病毒受体调节的见解。

PROJECT SUMMARY/ABSTRACT Adenoviruses (AdV) are common human pathogens that cause typical cold symptoms in healthy indi- viduals, but can potentially progress to acute respiratory distress syndrome (ARDS) with up to 50% mortality, particularly in highly susceptible, immunosuppressed people, and new, potentially lethal variants continue to emerge each year. No therapeutic that specifically prevents or treats AdV infection exists and the development of novel treatments would prevent the morbidity and potentially mortality associated with AdV infection. The objective of this proposal is to determine the mechanism of action and anti-adenovirus efficacy of novel molecules that decrease apical Coxsackievirus and adenovirus receptor (CAREx8) protein expression. Our central hypothesis is that decoy peptides that block the interaction between MAGI-1 and CAREx8 destabi- lize apical CAREx8 protein in the airway to abrogate AdV entry and pathogenesis. We will test our hypothesis using polarized model epithelia with Dox-inducible CAREx8 expression, well-differentiated primary human and rodent airway epithelia to validate our findings, and the cotton rat model to evaluate wildtype AdV pathogenesis in immunocompetent and cyclophosphamide-immunosuppressed animals, with two specific aims: Aim 1: To elucidate the mechanism of MAGI-1 activating peptide and AdV triggered proteolytic degra- dation of CAREx8. Completion of this aim will allow the identification and development of novel and much needed targets and approaches to prevent the infection and spread of pathogenic wild-type AdV and, in the future, group B coxsackieviruses. Aim 2: To define the protection afforded by MAGI-1 PDZ1 binding peptides against wild type adenovi- rus infection in cotton rats. Completion of this aim will provide proof-of-principle that AdV infection can be thwarted by reducing its primary receptor and may save lives of severely infected or immunosuppressed indi- viduals. Overall impact: Understanding the mechanisms that regulate the expression and localization of CAREx8, and how this unique isoform mediates viral entry at the apical surface of a polarized epithelium is criti- cal for understanding viral spread, tissue tropism, and pathogenesis. The successful completion of the pro- posed aims will identify not only the cellular mechanisms regulating the expression of the apical adenovirus receptor but also mechanisms regulating viral binding and infection. This will establish the feasibility of thera- peutic agents that reduce the susceptibility of the airway epithelium to adenovirus infection and pathogenicity prior to infection and during an active infection in immunocompetent and immunosuppressed conditions. We ultimately expect the proposed aims to lead to novel anti-viral interventions that may also block other viruses that use CAR as a primary receptor, and provide insight into the regulation of other related viral receptors.