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.

抽象的 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 在世界范围内具有显着的发病率和死亡率。这两种感染（一种病毒感染和一种细菌感染）有许多共同点 相同的传输特性；在美国，并发感染尤其普遍 危险人群，例如接受抗生素治疗的老年患者、住院患者或患者 住在长期护理机构中。尽管在开发针对这两种感染的疫苗方面付出了巨大努力， 迄今为止，市场上仍没有单一疫苗可用。我们开发了一种新颖的平台技术 通过改造益生菌酵母，布拉氏酵母菌，分泌多种细菌来对抗肠道细菌病原体 特异性单域（VHH）抗体，直接针对肠道部位的细菌毒力因子 感染。我们已成功地将这项技术应用于针对艰难梭菌。本次活动的总体目标 项目是生成针对 CD 毒素和 hNoV 分泌 mSdAb 的 Sb 菌株，并进行原理验证 相关动物疾病模型中的功效数据，并开发临床相容的干燥和干燥制剂 封装 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 将预防其单独或并发感染。为了检验这个假设， 我们建议实现以下 3 个具体目标：1）工程 Sb-mSdAb 菌株（Sb-aNoVCd）分泌 针对 CD 毒素和 hNoV 的 VHH 融合。 2) 确定 Sb-aNoVCd 菌株的预防功效 限生猪中的 hNoV 和 CD 感染。 3) 开发支持酵母喷雾干燥的配方 封装。随着拟议的转化活动的完成，我们将产生先导 Sb-aNoVCd 菌株并评估了小鼠和最合适动物的无菌猪的体内特性 hNoV感染和疾病模型。如果生成了关键的原理验证功效数据，未来的扩大规模 包括额外的 VHH 来产生广泛有效的先导免疫预防药物的努力将是合理的 菌株和针对 CD 和 hNoV 胃肠炎的酵母产品的最终商业化 目前没有预防措施或疫苗。