Oligonucleotide-based therapy for HUS

这项建议旨在开发针对溶血性尿毒症综合征(HUS)的治疗方法 由产志贺毒素(STX)的大肠杆菌(STEC)。STEC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。它们代表着一个重要的全球 与食源性疾病和潜在的生物恐怖主义有关的新感染。病毒的致命性 STEC的突出特点是产生临床疾病所需的极低感染剂量。这个 STEC感染的治疗因常规的潜在不良后果而变得复杂 抗生素的使用。腹泻相关(D+HUS)是STEC的一种危及生命的并发症 儿童和老年人感染，主要是O157：H7血清型，由突如其来的 出现面色苍白和少尿。它与显著的发病率有关，尽管在 儿科重症监护；这种疾病的死亡率保持在3-5%。目前还没有经过验证的治疗方法 对于HUS来说，这可以降低死亡率、急性透析的需要或严重肾外的发生 事件。最近的暴发，特别是菠菜的暴发，表明了 人口对意外污染的反应，以及防护或治疗措施的紧迫性 需要的。 在这一应用中，我们建议使用新开发的寡核苷酸/适配子技术，该技术 已经在人类的几种系统性疾病中获得了潜在的治疗剂。在这 应用程序两个团队通力合作，以实现此应用程序的目标。它们包括一个 在设计、合成和应用寡核苷酸/适体技术方面拥有专业知识的团队， 以及一支在STEC疾病和发育筛查以及体外和体内研究方面具有专业知识的团队 抗HUS治疗药物的评价。这项提议旨在检验这样一个假设： 针对Stx2和STX1专门设计的一个或几个适配子的管理将保护 小鼠和仔猪对全身STX攻击后致死性全身中毒的保护作用 口服大肠埃希菌O157攻击。具体目标1集中在合成、选择、 适配子的优化和修饰。特异性目标2将应用于体外检测 对体内研究中的适配子进行表征和排序。具体目标3将评估所选的 适配子在小鼠毒性模型中的疗效和药代动力学，并针对4个具体目标 在已建立的仔猪口服大肠杆菌O157：H7感染模型中进行临床前评估。

This 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 (D+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 oligonucleotide/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 oligonucleotide/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. This proposal is designed to test the hypothesis that administration of specifically designed one or several aptamers against Stx2 and Stx1 will protect mice and piglets against the fatal systemic intoxication induced following systemic Stx challenge and oral E. coli O157challenge, respectively. The Specific Aim 1 focuses on the synthesis, selection, optimization and modification of aptamers. The Specific Aim 2 will apply in vitro assays to characterize and rank aptamers for in vivo studies. The Specific Aim 3 will evaluate the selected aptamers in the mouse toxicity model for efficacy and pharmacokinetics, and the Specific Aim 4 will perform preclinical evaluation in the well-established piglet model of oral E. coli O157:H7 infection.