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: 10529090
• 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/10529090
• 关键词: Address; Animals; Bacteria; Bacteroides; Basic Science; Biochemical; Blood; Cell Culture Techniques; Chronic; Chronic Disease; Clinical; Colitis; Collagen; Colonic inflammation; Coupled; Crohn' s disease; Data Set; Development; Diagnosis; Disease; Disease Management; Epithelial; 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; base; 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）是一种慢性疾病，其特征是结肠粘膜炎症。 UC具有重大的全球负担，其特征是针对肠道微生物群的异常免疫反应。当前的治疗方案仅针对宿主炎症途径，并且在管理疾病方面通常无效。目前，北美的近100万个人居住在UC中，其在人类中的普遍性在全球范围内仍在增长。大量生长病例，再加上缺乏有效的疗法，强调需要调查针对UC的新治疗干预措施。我们的小组最近通过六个基于粪便或血清的 - 单个数据集，来自40名UC患者，表现出宿主 - 微生物组相互作用，这些数据集显示出广泛的临床，内窥镜和组织学疾病活性。经过大规模分析后，这六个数据集为中心假设提供了有力的证据：杆菌蛋白酶可以促进UC疾病的严重程度。元蛋白质组学将B. vulgatus蛋白酶定位为严重程度的独特特征。 shot弹枪宏基因组学指导了分类学的推断，并透露，vulgatus b。患者血清中发现的大量丝氨酸蛋白抑制剂表明蛋白酶的重要性。替肽组显示出与UC疾病严重程度相关的肽片段增加。一个独立的210人队列验证了B. vulgatus蛋白酶与UC疾病严重程度之间的牢固联系。为了检验我们的假设，我们证明了B. vulgatus可以在体外破坏肠上皮渗透性，并且蛋白酶抑制作用足以恢复上皮屏障。 Vulgatus在无毛发的IL-10缺乏小鼠中的单殖民化表现出这些动物的定殖，并且蛋白酶抑制作用足以防止结肠炎发育。此外，从UC患者中粪便过多的蛋白酶蛋白酶的粪便移植到无菌小鼠中诱导的结肠炎取决于蛋白酶的活性（Mills等人，自然微生物学2021）。根据我们发表的工作，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B. vulgatus利用的一种中心机制，可在体外诱导结肠炎和体内与UC和2）B. vulgatus蛋白酶降低了肠道障碍的关键成分。本应用的具体目的是定义基因组环境，角色并开发针对UC中的Vulgatus蛋白酶（AIM 1）的特定抑制剂（AIM 1），表征UC中Vulgatus介导的蛋白水解的靶标和动力学（AIM 2）。为了完成这项建议，我们将使用最先进的生化，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UC基础和翻译研究中脱颖而出。