Development of Aptamer-Based Therapy Against HUS

描述（由申请人提供）：本提案是根据RFA-AI-08-001提交的。我们的目标是开发针对由产滋贺毒素（Stx）的E. coli（STEC）。STEC菌株是严重的B类病原体，主要与食物和水传播获得性疾病相关。它们是一种重要的全球新出现的感染，与食源性疾病和潜在的生物恐怖主义有关。STEC的毒力突出表现为产生临床疾病所需的非常低的感染剂量。STEC感染的治疗因常规给予抗生素的潜在不良后果而复杂化。腹泻相关性（HUS）是STEC感染的一种危及生命的并发症，主要是O 157：H7血清型，在儿童和老年人中，其预示着突然出现苍白和少尿。它与显著的发病率相关，尽管儿科重症监护有所改善，但这种疾病的死亡率仍为3- 5%。目前尚无经证实的HUS治疗方法可降低死亡率、急性透析需求或严重肾外事件的发生率。最近的疫情，特别是菠菜疫情，表明人口是多么容易受到意外污染，以及多么迫切地需要保护或治疗措施。在本申请中，我们建议使用新开发的适体技术，该技术已在人类的几种全身性疾病中作为潜在的治疗剂获得了关注。在这个应用程序中，两个团队走到一起来实现这个应用程序的目标。他们包括一个在适体技术的设计、合成和应用方面具有专业知识的团队，以及一个在STEC疾病和针对HUS的治疗剂的开发筛选以及体外和体内评估方面具有专业知识的团队。因此：Specific Aim 1专注于适体的设计，合成和优化，Specific Aim 2将应用ELISA和基于细胞的检测来筛选适体。Specific Aim 3将在小鼠毒性模型中评价从基于细胞的筛选中产生的选定适体，包括药物毒性和药代动力学，而Specific Aim 4将在已建立的仔猪大肠杆菌模型中进行临床前评价和药代动力学。co//O 157：H7感染/全身中毒模型。相关性（参见说明）：在美国，儿童感染产滋贺毒素（Stx）大肠杆菌（STEC）是溶血性尿毒症综合征（HUS）的主要原因。目前还没有预防或改善HUS的特异性治疗方法。在本提案中，我们建议开发基于适体的治疗剂用于预防或治疗HUS。

DESCRIPTION (provided by applicant): This proposal is submitted in response to RFA-AI-08-001. Our proposal targets the development of therapy against hemolytic uremic syndrome (HUS) caused by Shiga toxin (Stx)-producing E. coli (STEC). STEC strains are serious Category B pathogens associated primarily with food and waterborne acquired disease. They represent an important global emerging infection with relevance to foodborne illness and potential bioterrorism. The virulence of STEC is underscored by the very low infectious dose required to produce clinical disease. The treatment of STEC infection is complicated by potentially adverse consequences of routine administration of antibiotics. Diarrhea-associated (HUS) is a life-threatening complication of STEC infection, primarily the O157:H7 serotype, in children and the elderly that is heralded by the sudden onset of pallor and oliguria. It is associated with significant morbidity and, despite improvements in pediatric intensive care; the mortality rate from this disease remains 3-5%. There is no proven therapy for HUS that reduces mortality, the need for acute dialysis, or the occurrence of serious extra-renal events. The recent outbreaks, the spinach outbreak in particular, showed how vulnerable the population is to accidental contamination, and how urgently protective or therapeutic measure are needed. In this application we propose to use the newly developed aptamer technology which has gained foot as potential therapeutic agents in several systemic diseases in humans. In this application two teams have come together to achieve the goals of this application. They include a team with expertise in the design, synthesis and application of aptamer technology, and a team with expertise on STEC disease and the development screening and in vitro and in vivo evaluation of therapeutic agents against HUS. Consequently: Specific Aim 1 focuses on the design, synthesis and optimization of aptamers, Specific Aim 2 will apply ELISA and cell-based assays for screening of aptamers. Specific Aim 3 will evaluate in the mouse toxicity model selected aptamers generated from the cell based screening, including drug toxicity and pharmacokinetics, while Specific Aim 4 will perform preclinical evaluation and pharmacokinetics in the well-established piglet model of E. co//O157:H7 infection/systemic intoxication model. RELEVANCE (See instructions): Infection of children with Shiga toxin (Stx) producing Escherichia coli (STEC), primarily a food and waterborne acquired disease, is the leading cause of hemolytic-uremic syndrome (HUS) in the US. There is no specific treatment to prevent or ameliorate HUS. In this proposal, we propose to develop aptamer-based therapeutic agents for prevention or treatment of HUS.