Therapeutics to Prevent/Treat Lassa Fever Virus

描述（由申请人提供）：病毒要感染细胞并随后引起疾病，必须首先与细胞受体结合。Oldstone实验室分离并鉴定了几种沙病毒（包括拉沙热病毒（LFV）和LCMV CI 13）的细胞受体α -三磷酸甘聚糖(（-DG）。我们提供的证据表明(-DG是感染所需的受体（Cao et al., Science, 1998; Spiropoulou et al.， J. Virol）。， 2002)，因为：1)LFV和LFV糖蛋白（GP）与固定在膜上的纯化（-DG）有高亲和力结合；2)DG基因零突变的正常允许细胞对LFV感染具有抗性；3)利用腺病毒载体重建（-DG）在零突变细胞中的表达，恢复了对LFV的易感性。我们建立了一种高输出量的方法来分析BSL2条件下lfvgp介导的靶细胞感染。在Boger实验室，液相合成技术已被用于创建独特的小分子组合文库，这些小分子可以被筛选以识别促进蛋白质-蛋白质相互作用（激动剂）或抑制蛋白质-蛋白质相互作用（拮抗剂）的治疗性化合物。

DESCRIPTION (provided by applicant): For a virus to infect a cell and subsequently cause disease it must first bind to its cellular receptor. The Oldstone laboratory isolated and characterized the cellular receptor, alpha-dystroglycan ((-DG) for several arenaviruses including Lassa fever virus (LFV) and LCMV CI 13. We provided evidence that (-DG is the receptor required for infection (Cao et al., Science, 1998; Spiropoulou et al., J. Virol., 2002) as: 1) LFV and LFV glycoprotein (GP) bind at high affinity to purified (-DG immobilized on membranes; 2) normally permissive cells bearing a null mutation of the DG gene are resistant to LFV infection; 3) reconstitution of (-DG expression in null mutant cells using an adenovirus vector restored susceptibility to LFV. We have established a high-output assay to analyze LFVGP-mediated infection of target cells under BSL2 conditions. In the Boger laboratory solution-phase synthetic techniques have been utilized to create unique combinatorial libraries of small molecules that can be screened to identify therapeutic compounds that promote protein-protein interactions (agonists) or inhibit protein-protein interactions (antagonists). We propose three specific aims: First, to screen combinatorial chemical libraries to discover small molecules that inhibit LFVGP-mediated infection of cells using retroviral vectors that contain LFVGP in their envelope and a green fluorescent protein reporter gene. Second, to develop a drug that would neutralize free LFV by engineering anti-viral receptor-bodies, in which the Fab part of an IgG molecule is replaced by a virus-binding (-DG fragment. Third, to validate this inhibition in vivo by blocking LCMV CI 13 infection in a mouse model (CI 13 and LFV binds at high affinity to (-DG and predictably utilizes the same binding site). Final verification would be done at CDC using the experimentally defined chemical inhibitor(s) and virulent LFV.