Mechanisms of Enterovirus Entry

利用高分辨率冷冻电子显微镜(Cryo EM)，我们将研究柯萨奇病毒B3(CVB3)通过与柯萨奇病毒和腺病毒受体(CAR)结合进入宿主细胞的机制。CVB3是一种引起心肌炎、胰腺炎的人类病原体，最近被认为与青少年糖尿病的发病有关。我们提出的关于CVB3进入的研究是对先前富有成效的授权期的延续。之前的部分工作是与费城儿童医院传染病科杰弗里·伯格尔森博士合作完成的，伯格森博士现已退休。继续研究肠道病毒进入的拟议项目包括与FDA康斯坦丁·楚马科夫合作。Chumakov将与我们合作探索前一个资助期最有希望的结果之一，即发现CVB3衣壳上的汽车足迹区域在其他三种病毒及其受体中是保守的，从而使用三种不同的宿主蛋白进入四个物种。进一步检查表明，1)该区域在所有人类肠道病毒中高度保守，包括所有三种脊髓灰质炎病毒株。2)共有四个不同的高度保守区。以前的工作和我们的初步结果将这些保守的站点与与条目相关的功能联系起来。我们的结构/功能研究将包括测试每个区域的功能，并将其作为迫切需要的抗病毒药物的靶标。我们将使用传统的二十面体平均方法以及不对称重建方法，其中一些方法是我们最近开发的。

Using high resolution cryo electron microscopy (Cryo EM) we will investigate the mechanisms used by coxsackievirus B3 (CVB3) to enter host cells by engaging the coxsackievirus and adenovirus receptor (CAR). CVB3 is a human pathogen that causes myocarditis, pancreatitis, and has recently been linked to the onset of juvenile diabetes mellitus. Our proposed studies of CVB3 entry are a continuation of a productive previous granting period. Part of the previous work done was in collaboration with Dr. Jeffery Bergelson, Division of Infectious Diseases at Children's Hospital at Philadelphia, who has since retired. The proposed projects to continue studies of enterovirus entry include a collaboration with Konstantin Chumakov, FDA. Chumakov will work with us to explore one of the most promising results from the previous funding period, which was the discovery that a region in the CAR footprint onto the CVB3 capsid is conserved among three other viruses and their receptors, thus crossing four species using three different host proteins for entry. Further examination showed 1) the region is highly conserved among all human enteroviruses, including all three strains of poliovirus. 2) there are in total four different highly conserved regions. Previous work and our preliminary results link these conserved sites to functions linked to entry. Our structure/function studies will include testing each region for function and as targets for antivirals that are desperately needed. We will use both traditional icosahedrally averaged approaches along with asymmetric reconstruction approaches, some of which have been recently developed by us.