Inflammatory Effect of Enteric Bacterial-Mediated Intestinal Permeability

肠道细菌介导的肠道通透性的炎症效应

• 批准号: 8331442
• 负责人: Ian Michael Carroll
• 金额: $ 11.94万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-15 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8331442
• 关键词: Address; Adverse effects; Affect; Antibiotics; Award; Bacteria; Bifidobacterium; Biological; Caring; Catalytic Domain; Cells; Colitis; Collaborations; Comorbidity; Complex; Crohn' s disease; Disease; Enteral; Enterobacteriaceae; Enterococcus faecalis; Epithelial; Epithelial Cells; Functional disorder; Gastrointestinal Diseases; Gelatinases; Germ-Free; Gnotobiotic; Goals; Heterogeneity; Human; Immune system; In Vitro; Individual; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Inflammatory disease of the intestine; Interleukin-10; Intestinal Diseases; Intestines; Knowledge; Lead; Measures; Mediating; Medical; Mentors; Mono-S; Mus; Outcome; PAR-1 Receptor; Patients; Peptide Hydrolases; Permeability; Pharmaceutical Preparations; Physiology; Probiotics; Proteinase-Activated Receptors; Public Health; Quality of life; Receptor Activation; Recurrence; Research; Resources; Serine; Serine Protease; Serine Proteinase Inhibitors; Serpins; Serum; Source; Testing; Therapeutic; Tight Junctions; Training; Treatment Cost; Ulcerative Colitis; United States; Wild Type Mouse; clinical practice; design; duodenitis; effective therapy; in vivo Model; innovation; meetings; member; microbial; monolayer; mutant; new therapeutic target; novel; novel therapeutic intervention; psychologic; success; therapeutic target

DESCRIPTION (provided by applicant): Inflammatory bowel diseases (IBD) are highly prevalent intestinal diseases in the United States affecting 1.4 million individuals. These diseases are associated with reduced quality of life and psychological co-morbidity. Current estimates for IBD associated treatment costs in the US are $6.3 billion. The high rate of recurrence and lack of safe and curative treatments for IBD underscore the need for alternate therapeutic approaches for these complex diseases. Bacterial proteases capable of inducing intestinal permeability via protease-activated receptors (PARs) present a novel therapeutic target for IBD. The goals for this proposal are to (I) elucidate the specific mechanism by which enteric bacteria induce intestinal permeability, (II) assess the biological effect of protease producing and inhibiting enteric bacteria on intestinal permeability, and (III) assess the inflammatory effect of protease producing and inhibiting enteric bacteria on the intestine. To address these goals we have proposed three specific aims. In specific aim 1 we will determine the ability of protease-producing and - inhibiting enteric bacterial strains to regulate PARs and permeability in human epithelial cells in vitro. To achieve this aim we will use wild-type and mutant Enterococcus faecalis OG1RF strains that lack gelatinase or serine protease activity, and wild-type and mutant Bifidobacterium longum ATCC15707 strains that lack serpin activity. We will expose tight junction forming T-84 epithelial cell monolayers to parental and mutant E. faecalis OG1RF and B. longum ATCC15707 strains and measure cell permeability and PAR activation. In specific aim 2 we will determine the ability of protease-producing and inhibiting enteric bacterial strains to regulate PAR- dependent intestinal permeability in gnotobiotic mice. To achieve this aim we will mono- and dual-associate germ-free wild-type and PAR deficient mice with parental and mutant bacterial strains and measure intestinal permeability and PAR activation. In specific aim 3 we will determine the ability of protease-producing and inhibiting enteric bacterial strains to induce intestinal permeability and inflammation in gnotobiotic IL-10 deficient (IL-10-/-) mice. To achieve this aim we will mono- and dual-associate germ-free wild-type and IL-10-/- mice with parental and mutant bacterial strains and measure intestinal permeability, PAR activation, and inflammation. The contribution of the proposed research is significant as it will define a precise mechanism by which enteric bacteria alter intestinal permeability and contribute to inflammation. The proposed research will also be of significance because the outcome will contribute to the broader understanding of mechanisms by which dysbiosis in the intestinal microbiota affects IBD. Additionally, the proposed research is innovative in our opinion as it will allow for the design of rational pathophysiology-directed probiotic treatment of GI disease.

描述（由申请人提供）：炎症性肠病（IBD）是美国影响140万人的高度普遍肠道疾病。这些疾病与生活质量降低和心理疾病有关。美国目前对美国相关治疗费用的估计为63亿美元。 IBD的高复发率和缺乏安全和治疗方法强调了对这些复杂疾病的替代治疗方法的需求。能够通过蛋白酶激活受体诱导肠道通透性（PAR）的细菌蛋白酶为IBD提供了一种新型的治疗靶标。该建议的目标是（i）阐明肠细菌诱导肠道通透性的特定机制，（ii）评估蛋白酶产生和抑制肠细菌对肠道通透性的生物学作用，以及（iii）评估蛋白酶产生蛋白酶的炎症和抑制肠细胞质对肠道的炎症。为了解决这些目标，我们提出了三个具体目标。在特定目标1中，我们将确定产生蛋白酶的能力和 - 抑制肠细菌菌株在体外调节人上皮细胞中PAR和渗透性的能力。为了实现这一目标，我们将使用缺乏明胶酶或丝氨酸蛋白酶活性的野生型和突变体肠球菌OG1RF菌株，以及缺乏Serpin活性的野生型和突变的双歧杆菌ATCC15707菌株。我们将暴露于形成T-84上皮细胞单层的紧密连接到父母和突变的E. faecalis og1rf和B. longum atcc15707菌株，并测量细胞的渗透性和PAR激活。在特定的目标2中，我们将确定产生蛋白酶和抑制肠细菌菌株的能力，以调节gnotobiotic小鼠的肠道肠道通透性。为了实现这一目标，我们将与父母和突变细菌菌株单一和双缔合无菌野生型和PAR缺陷小鼠，并测量肠道通透性和PAR激活。在特定的目标3中，我们将确定产生蛋白酶和抑制肠细菌菌株诱导肠道通透性和炎症的能力。为了实现此目的，我们将单一和双缔合无菌野生型和IL-10 - / - 小鼠具有父母和突变细菌菌株，并测量肠道通透性，PAR激活和炎症。拟议的研究的贡献很重要，因为它将定义肠细菌改变肠道通透性并导致炎症的精确机制。拟议的研究也将具有重要意义，因为结果将有助于对肠道菌群中营养不良影响IBD的机制的更广泛的理解。此外，在我们看来，拟议的研究具有创新性，因为它将允许设计理性的病理生理学指导的益生菌治疗GI疾病。