Preventing norovirus and Clostridium difficile gastroenteritis by engineered probiotic yeast Saccharomyces boulardii secreting multi-specific single-domain antibodies

通过分泌多特异性单域抗体的工程益生菌布拉酵母菌预防诺如病毒和艰难梭菌胃肠炎

• 批准号: 10320907
• 负责人: Hanping Feng
• 金额: $ 60.86万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-22 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10320907
• 关键词: Acute; Animal Disease Models; Animal Model; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Antibodies; Binding; Body Weight decreased; Carbohydrates; Characteristics; Chromosomal Insertion; Clinic; Clinical; Clostridium difficile; Communicable Diseases; DNA cassette; Data; Development; Diarrhea; Disease; Dose; Dried Yeast; Drug or chemical Tissue Distribution; Encapsulated; Engineered Probiotics; Engineering; Enteral; Erythrocytes; Excipients; Family suidae; Formulation; Future; Gastroenteritis; Genes; Genome; Gnotobiotic; Goals; Hand; Health care facility; Hemagglutination; Human; In Vitro; Individual; Infection; Infection prevention; Inflammation; Intestinal Secretions; Intestines; Lead; Long-Term Care; Mediating; Methods; Monitor; Morbidity - disease rate; Mus; Norovirus; Oral; Oral Administration; Patients; Prevention strategy; Preventive; Probiotics; Process; Production; Prophylactic treatment; Saccharomyces; Site; Symptoms; Technology; Testing; Therapeutic; Toxin; Traveler' s diarrhea; Vaccines; Variant; Viral; Viral Load result; Virulence Factors; Virus-like particle; Yeasts; aged; antibiotic-associated diarrhea; antitoxin; base; capsule; commercialization; enteric pathogen; gut microbiota; high risk population; in vivo; microbiota; microbiota transplantation; mortality; nanobodies; novel; passive antibodies; pathogen; pathogenic bacteria; prevent; scale up; tool; transmission process; vaccine access; vaccine development

Abstract Human norovirus (hNoV) and Clostridium difficile (CD) represent two leading causes of acute gastroenteritis worldwide with significant morbidity and mortality. The two infections (one viral and one bacterial) share many of the same characteristics of transmission; and concurrent infections are particularly prevalent in the US in high- risk populations, such as aged patients undergoing antibiotic treatments, hospitalized patients, or patients staying in long-term care facilities. Despite great efforts in the development of vaccines against both infections, to date there is still no single vaccine available on the market. We have developed a novel platform technology against enteric bacterial pathogens by engineering a probiotic yeast, Saccharomyces boulardii, to secrete multi- specific single-domain (VHH) antibodies, directly targeting bacterial virulence factors at the intestinal site of infection. We have successfully applied this technology to target Clostridium difficile. The overall objective of this project is to generate Sb strains secreting mSdAbs against both CD toxins and hNoV, generate proof-of-principle efficacy data in relevant animal disease models, and develop clinic-compatible formulations for drying and encapsulating the Sb-mSdAb strains. We hypothesize that oral administration of Sb-mSdAb strains secreting mSdAb against hNoV and CD toxins will prevent their individual or concurrent infections. To test this hypothesis, we propose to accomplish the following 3 specific aims: 1) Engineer Sb-mSdAb strains (Sb-aNoVCd) secreting VHH fusions against both CD toxins and hNoVs. 2) Determine preventive efficacy of Sb-aNoVCd strains against infections with hNoV and CD in gnotobiotic pigs. 3) Develop a formulation supporting yeast spray drying and encapsulation. With the completion of proposed translational activities, we will have generated lead Sb-aNoVCd strains and evaluated in vivo characteristics in mice and in gnotobiotic pigs which is the most appropriate animal model of hNoV infection and disease. Should the key proof-of-principle efficacy data generated, future scale-up efforts will be justified to include additional VHHs for generating broadly efficacious lead immunoprophylactic strains and eventual commercialization of the yeast products against CD and hNoV gastroenteritis for which we currently have no prophylaxis or vaccines.

摘要 人类诺如病毒(Hnov)和艰难梭菌(Cd)是急性胃肠炎的两个主要原因。 世界各地都有严重的发病率和死亡率。这两种感染(一种病毒和一种细菌)有许多共同之处 传播的相同特征；同时感染在美国尤其普遍，在高 危险人群，如接受抗生素治疗的老年患者、住院患者或患者 住在长期护理机构。尽管在开发针对这两种感染的疫苗方面做出了很大努力， 到目前为止，市场上仍然没有单一的疫苗可用。我们开发了一种新的平台技术 通过工程设计一种益生菌酵母来对抗肠道细菌病原体，该酵母菌可以分泌多个 特异性单域(VHH)抗体，直接靶向肠道部位的细菌毒力因子 感染。我们已经成功地将该技术应用于艰难梭菌的靶向。这样做的总体目标是 项目是产生分泌抗Cd毒素和hNoV的mSdAb的SB菌株，产生原理证明 相关动物疾病模型的疗效数据，并开发临床兼容的干燥和 包被Sb-mSdAb菌株。我们假设口服Sb-mSdAb菌株分泌 针对hNoV和CD毒素的mSdAb将防止其单独或同时感染。为了检验这一假设， 我们提出了以下三个具体目标：1)工程SB-mSdAb株(SB-aNoVCd)分泌 针对Cd毒素和hNov的VHH融合。2)测定SB-aNoVCd菌株的预防效果 人新城疫病毒和环糊精在诺生菌猪体内的感染。3)开发支持酵母喷雾干燥的配方， 封装。完成建议的翻译活动后，我们将产生销售线索SB-aNoVCd 菌株及其在小鼠和灵知生菌猪体内的特性，这是最适合的动物 人新城疫病毒感染和疾病模型。关键的原则性疗效数据是否应该生成，未来扩大 将有理由将更多的VHH纳入产生广泛有效的铅免疫预防措施 抗CD和hNOV胃肠炎酵母菌的菌株和最终商品化 目前还没有预防措施或疫苗。