Microbiome Driven Proteolysis as a Contributing Factor to Severity of Ulcerative Colitis Disease Activity

微生物组驱动的蛋白水解是溃疡性结肠炎疾病活动严重程度的影响因素

• 批准号: 10673152
• 负责人: David J Gonzalez
• 金额: $ 54万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10673152
• 关键词: Address; Animals; Bacteria; Bacteroides; Basic Science; Biochemical; Blood; Cell Culture Techniques; Chronic; Chronic Disease; Clinical; Colitis; Collagen; Colon; Colonic inflammation; Coupled; Crohn' s disease; Data Set; Development; Diagnosis; Disease; Disease Management; Epithelium; Feces; Functional disorder; Genetic; Genomics; Germ-Free; Goals; Histologic; Human; Immune response; In Vitro; Individual; Inflammatory; Inflammatory Bowel Diseases; Integration Host Factors; Interleukin-10; Intestinal permeability; Investigation; Link; Mediating; Metagenomics; Microbiology; Modeling; Mucins; Mucositis; Mus; Nature; North America; Pathway interactions; Patients; Peptide Fragments; Peptide Hydrolases; Persons; Play; Prevalence; Protease Inhibitor; Proteins; Proteolysis; Publishing; Regulation; Role; Sampling; Serine Proteinase Inhibitors; Serum; Severities; Severity of illness; Shotguns; Stress; Structure; Taxonomy; Testing; Therapeutic; Tight Junctions; Translational Research; Transplantation; Ulcerative Colitis; Validation; Work; bacterial genetics; cohort; data integration; design; effective therapy; gut microbiome; gut microbiota; host microbiome; host microbiota; improved; in vitro Model; in vivo; in vivo Model; inhibitor; intestinal barrier; metaproteomics; microbial; microbiome; microbiome research; microbiota; mutant; novel; novel strategies; novel therapeutic intervention; prevent; tool

Ulcerative colitis (UC) is a chronic disease characterized by inflammation of the mucosa of the colon. UC has a significant global burden, and is characterized by an aberrant immune response directed towards the gut microbiota. Current treatment options exclusively target host inflammatory pathways and are often ineffective in managing disease. Nearly 1 million individuals in North America are currently living with UC and its prevalence in humans continues to grow worldwide. The considerable number of growing cases, coupled with the lack of an effective therapy, stresses the need for investigation into new therapeutic interventions against UC. Our group recently characterized host-microbiome interactions governing UC through six fecal or serum based –omic datasets from 40 UC patients displaying a wide range of clinical, endoscopic, and histologic disease activity. After broad-scale analyses, the six datasets provided powerful evidence towards a central hypothesis: Bacteroides vulgatus proteases can drive UC disease severity. Metaproteomics pinpointed B. vulgatus proteases as a distinguishing feature of severity. Shotgun metagenomics guided taxonomic inferences and revealed that the B. vulgatus association was driven primarily by protein regulation as opposed to microbial abundances. An abundance of serine protease inhibitors found in the patient serum suggested the importance of proteases. The metapeptidome showed increased peptide fragments correlated with UC disease severity. An independent 210-person cohort validated the strong connection between B. vulgatus proteases and UC disease severity. Testing our hypothesis, we demonstrate B. vulgatus can disrupt intestinal epithelial permeability in vitro, and protease inhibition was sufficient to restore epithelial barrier. Monocolonization of B. vulgatus into germfree IL-10 deficient mice demonstrated colonization induces colitis in these animals and protease inhibition is sufficient to prevent colitis development. Furthermore, transplantation of feces from UC patients with over-abundant B. vulgatus proteases into germ-free mice induced colitis dependent on protease activity (Mills et al., Nature Microbiology 2021). Based on our published work, we have formulated two testable hypotheses to further build on these new findings: 1) proteolysis is a central mechanism utilized by B. vulgatus to induce colitis in vitro and in vivo relevant to UC and 2) B. vulgatus proteases degrade key components of the intestinal barrier. The specific aims of this application are to define the genomic context, roles, and develop specific inhibitors against B. vulgatus proteases in UC (Aim 1), characterize the targets and dynamics of B. vulgatus mediated proteolysis in UC (Aim 2). To complete this proposal, we will use a multifaceted approach of state-of-the-art biochemical, cell culture, bacterial genetics, and in vivo models. Ultimately, this work has the potential to break new ground in UC basic and translational research.

溃疡性结肠炎（UC）是一种以结肠粘膜炎症为特征的慢性疾病。 UC 具有显着的全球负担，其特点是针对肠道微生物群的异常免疫反应。目前的治疗方案专门针对宿主炎症途径，并且通常在控制疾病方面无效。目前北美有近 100 万人患有 UC，并且其在全球人类中的患病率持续增长。大量不断增长的病例，加上缺乏有效的治疗方法，强调需要研究针对 UC 的新治疗干预措施。我们的小组最近通过来自 40 名 UC 患者的 6 个基于粪便或血清的组学数据集来表征控制 UC 的宿主-微生物组相互作用，这些患者显示了广泛的临床、内窥镜和组织学疾病活动。经过大规模分析后，这六个数据集为一个中心假设提供了有力的证据：普通拟杆菌蛋白酶可以导致 UC 疾病的严重程度。宏蛋白质组学将普通芽孢杆菌蛋白酶确定为严重程度的一个显着特征。鸟枪法宏基因组学指导了分类学推断，并揭示了普通拟杆菌的关联主要是由蛋白质调节驱动的，而不是微生物丰度。在患者血清中发现的大量丝氨酸蛋白酶抑制剂表明了蛋白酶的重要性。元肽组显示与 UC 疾病严重程度相关的肽片段增加。一个由 210 人组成的独立队列验证了普通双歧杆菌蛋白酶与 UC 疾病严重程度之间的密切联系。检验我们的假设，我们证明普通双歧杆菌可以在体外破坏肠上皮通透性，并且蛋白酶抑制足以恢复上皮屏障。将普通双歧杆菌单定植到无菌的 IL-10 缺陷小鼠体内，证明定植会在这些动物中诱发结肠炎，并且蛋白酶抑制足以预防结肠炎的发展。此外，将含有过量普通芽孢杆菌蛋白酶的 UC 患者的粪便移植到无菌小鼠体内会诱发依赖于蛋白酶活性的结肠炎（Mills 等人，Nature Microbiology 2021）。基于我们已发表的工作，我们提出了两个可检验的假设，以进一步建立在这些新发现的基础上：1) 蛋白水解是普通双歧杆菌用来在体外和体内诱导与 UC 相关的结肠炎的核心机制；2) 普通双歧杆菌蛋白酶降解肠道屏障的关键成分。本申请的具体目的是定义 UC 中寻常双歧杆菌蛋白酶的基因组背景、作用并开发特异性抑制剂（目标 1），表征 UC 中寻常双歧杆菌介导的蛋白水解作用的靶标和动态（目标 2）。为了完成这项提案，我们将采用最先进的生化、细胞培养、细菌遗传学和体内模型的多方面方法。最终，这项工作有可能在加州大学基础研究和转化研究中开辟新天地。