Development of Aptamer-Based Therapy Against HUS

基于适体的 HUS 疗法的开发

• 批准号: 7929518
• 负责人: SAUL r TZIPORI
• 金额: $ 66.47万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-12 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7929518
• 关键词: Acute; Adverse effects; Affinity; Animals; Antibiotics; Aptamer Technology; Binding; Biochemical; Biological Assay; Bioterrorism; Blood Circulation; Categories; Cells; Child; Childhood; Clinical; Complication; DNA; Development; Dialysis procedure; Diarrhea; Disease; Disease Outbreaks; Dose; Drug Kinetics; Drug toxicity; Elderly; Enzyme-Linked Immunosorbent Assay; Escherichia coli; Escherichia coli EHEC; Escherichia coli O157; Evaluation; Event; Generations; Gnotobiotic; Goals; Half-Life; Hela Cells; Hemolytic-Uremic Syndrome; Human; Immunoglobulin Variable Region; In Vitro; Infection; Instruction; Intensive Care; Intoxication; Kidney; Lead; Life; Measures; Modeling; Morbidity - disease rate; Mus; Mutagenesis; Oligonucleotides; Oliguria; Oral; Pallor; Penetration; Polymers; Prevention; Protein Biosynthesis; Protein Synthesis Inhibition; Receptor Cell; Screening procedure; Serotyping; Serum; Shiga Toxin; Spinach - dietary; Systemic disease; Systemic infection; Testing; Therapeutic; Therapeutic Agents; Tissues; Toxic effect; Toxin; Variant; Virulence; Vulnerable Populations; adverse outcome; aptamer; base; cytotoxicity; design; foodborne; foodborne illness; foot; improved; in vitro Assay; in vivo; molecular size; mortality; pathogen; preclinical evaluation; prevent; protective efficacy; response; therapy development; uptake; waterborne

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.

说明(申请人提供)：本建议书是根据RFA-AI-08-001的要求提交的。我们的建议旨在开发治疗由产志贺毒素(STX)的大肠杆菌(STEC)引起的溶血性尿毒症综合征(HUS)的方法。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。它们代表着与食源性疾病和潜在的生物恐怖主义有关的一种重要的全球新感染。STEC的毒力因产生临床疾病所需的极低感染剂量而得到强调。STEC感染的治疗因常规使用抗生素可能产生的不良后果而变得复杂。腹泻相关(HUS)是STEC感染的一种危及生命的并发症，主要是O157：H7血清型，在儿童和老年人中，以突然出现苍白和少尿为先兆。它与显著的发病率有关，尽管儿科重症监护有所改善，但这种疾病的死亡率仍保持在3-5%。目前尚无有效的治疗HUS的方法可以降低死亡率、减少急性透析的需要或减少严重的肾外事件的发生。最近的疫情，特别是菠菜的疫情，表明了人口对意外污染的脆弱性，以及迫切需要采取保护或治疗措施。在这项应用中，我们建议使用新开发的适体技术，该技术已经在人类几种系统性疾病中作为潜在的治疗剂。在此应用程序中，两个团队联合起来以实现此应用程序的目标。他们包括一个在适体技术的设计、合成和应用方面拥有专业知识的团队，以及一个在STEC疾病以及针对HUS的治疗剂的开发筛查和体外和体内评估方面的专业团队。因此：特定目标1侧重于适配子的设计、合成和优化，特定目标2将应用ELISA法和基于细胞的分析来筛选适配子。特异性目标3将在小鼠毒性模型中评估基于细胞筛选产生的选定适配子，包括药物毒性和药代动力学，而特异性目标4将在已建立的E.co//O157：H7感染/全身中毒模型的仔猪模型中进行临床前评估和药代动力学。相关性(见说明)：儿童感染志贺毒素(STX)产生大肠杆菌(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。目前还没有专门的治疗方法来预防或改善HUS。在这项提案中，我们建议开发基于适体的治疗试剂来预防或治疗HUS。