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。我们的建议是针对由产志贺毒素（Stx）的大肠杆菌（STEC）引起的溶血性尿毒症综合征（HUS）的治疗发展。产大肠杆菌菌株是严重的B类病原体，主要与食物和水传播的获得性疾病有关。它们是一种重要的全球新发感染，与食源性疾病和潜在的生物恐怖主义有关。产志毒素大肠杆菌的毒力因产生临床疾病所需的极低传染剂量而得到强调。常规使用抗生素的潜在不良后果使产志贺毒素大肠杆菌感染的治疗复杂化。腹泻相关（HUS）是产血性大肠杆菌感染的一种危及生命的并发症，主要是O157:H7血清型，发生在儿童和老年人中，其先兆是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；这种疾病的死亡率仍为3-5%。目前尚无经证实的溶血性尿毒综合征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。最近的疫情，特别是菠菜疫情，表明了人群对意外污染的脆弱性，以及迫切需要采取的保护或治疗措施。在此应用中，我们建议使用新开发的适体技术，该技术已成为几种人类全身性疾病的潜在治疗剂。在这个应用程序中，两个团队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，以及一个在产志贺毒素大肠杆菌疾病和开发筛选以及针对溶血性尿毒综合征治疗剂的体外和体内评估方面具有专业知识的团队。因此，Specific Aim 1侧重于适配体的设计、合成和优化，Specific Aim 2将应用ELISA和基于细胞的检测来筛选适配体。Specific Aim 3将在小鼠毒性模型中评估从细胞筛选产生的适体，包括药物毒性和药代动力学，而Specific Aim 4将在建立的猪e.o //O157:H7感染/全身中毒模型中进行临床前评估和药代动力学。相关性（见说明）：儿童感染产生志贺毒素（Stx）的大肠杆菌（STEC），主要是一种通过食物和水传播的获得性疾病，是美国溶血性尿毒症综合征（HUS）的主要原因。没有专门的治疗方法来预防或改善溶血性尿毒综合征。在此建议中，我们建议开发基于适配体的治疗药物来预防或治疗溶血性尿毒综合征。

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.