Mucin-degrading microflora for prophylactic antibiotics

描述（由申请人提供）：该重新提交的I期SBIR申请的重点是开发一种粘蛋白降解结肠植物群，该菌群竞争性排除艰难梭菌，艰难梭菌是一种革兰氏阳性厌氧菌，可引起从轻度腹泻到危及生命的假膜性结肠炎等肠道疾病。我们的目标是C。因为这种病原体已经是美国的一个主要医疗问题，每年超过30万例，复发率接近20%。在大规模的抗菌预防措施的情况下，将有一个更大的人口高度容易感染C。很难C.艰难梭菌也可以被遗传操作以表达其它毒素，例如选择剂、肉毒杆菌毒素和肉毒杆菌毒素。以往的研究表明，球状梭菌群的成员降解和利用粘蛋白，并竞争性地排除C。从肠道中分离出来。在目标1中，我们将鉴定和表征来自球孢子菌群的粘蛋白降解植物群的潜在成员。我们将确定最有效地代谢粘蛋白的球菌组成员。将建立最佳粘蛋白降解菌株的生长和维持程序。在目标2中，我们将确定粘蛋白降解植物群的体外和体内性能特征。将评价植物群在仓鼠腹泻和结肠炎模型中的保护和治疗功效。将筛选有效植物群的成员，以确定是否不存在可转移的抗生素抗性基因和毒素基因。在第2阶段，我们的研究将扩展到生产GMP下的粘蛋白降解植物群，并在杜克大学Kenneth Wilson博士的指导下启动植物群的临床研究。我们的目标是确定粘蛋白降解植物群是否恢复或保持正常的粘蛋白代谢，增强对C. difficile，并降低C.艰难梭菌引起的腹泻

DESCRIPTION (provided by applicant): This resubmitted Phase I SBIR application focuses on the development of a mucin-degrading colonic flora that competitively excludes Clostridium difficile, a gram-positive anaerobe that causes antibiotic-associated intestinal disease ranging from mild diarrhea to life-theatening pseudomembranous colitis. We are targeting C. difficile because this pathogen already is a major healthcare problem in the U.S., with over 300,000 cases per year and a relapse rate approaching 20%. In the event of massive antimicrobial prophylaxis, there will be a much larger population highly susceptible to infection with C. difficile. C. difficile also could be genetically manipulated to express other toxins such as the select agents, epsilon and botulinum toxins. Previous studies have shown that members of the Clostridium coccoides group degrade and utilize mucin, and competitively exclude C. difficile from the intestine. In Aim 1, we will identify and characterize potential members of a mucin-degrading flora from the coccoides group. We will identify the coccoides group members that most effectively metabolize mucins. The growth and maintenance procedures for the top mucin-degrading strains will be established. In Aim 2, we will determine the in vitro and in vivo performance characteristics of a mucin-degrading flora. The flora will be evaluated for protection and treatment efficacy in the hamster model of antibiotic-associated diarrhea and colitis. Members of an effective flora will be screened for the absence of transferable antibiotic resistance genes and toxin genes. In Phase 2, our studies will be extended to produce the mucin-degrading flora under GMP and initiate clinical studies with the flora under the direction of Dr. Kenneth Wilson at Duke University. Our goal is to determine if the mucin-degrading flora restores or preserves normal mucin metabolism, enhances resistance to colonization by C. difficile, and reduces the incidence of C. difficile- induced antibiotic-associated diarrhea.