Polymicrobial interactions in Crohn's Disease

克罗恩病中的多种微生物相互作用

• 批准号: 9973148
• 负责人: Mahmoud A Ghannoum
• 金额: $ 61.74万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-08 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9973148
• 关键词: Address; Adhesions; Affect; Anaerobic Bacteria; Anti-Bacterial Agents; Antifungal Agents; Bacteria; Candida; Candida albicans; Candida tropicalis; Chronic; Clinical Research; Coculture Techniques; Colitis; Colon; Communities; Crohn' s disease; Data; Diagnostic; Environment; Escherichia coli; Exhibits; Exposure to; Family; Feces; Filament; Flare; Gastrointestinal tract structure; Gene Expression; Generations; Genes; Goals; Habits; Hyphae; Immune response; In Vitro; Indoles; Infection; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Interruption; Intestinal Mucosa; Intestines; Knock-out; Lactobacillus; Letters; Lymphoid Cell; Mediating; Microbe; Microbial Biofilms; Modeling; Modification; Mucous Membrane; Mus; Oral; Organism; Outcome; Pathogenicity; Patients; Phase; Phenotype; Play; Prevalence; Process; Production; Pyrrolidonecarboxylic Acid; Recovery; Relapse; Reporting; Resolution; Role; Saccharomyces; Sampling; Serratia marcescens; Severities; Sodium Dextran Sulfate; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Symptoms; Testing; Therapeutic; Thick; Tissues; Trichosporon; Tryptophan; Ulcerative Colitis; Virulence Factors; Yeasts; aryl hydrocarbon receptor ligand; bacteriome; design; dextran sulfate sodium induced colitis; experimental study; fungus; gut microbiota; healing; host microbiota; in vivo; in vivo evaluation; in vivo imaging; indoleacetic acid; inflammatory disease of the intestine; inhibitor/antagonist; insight; interleukin-22; intestinal injury; metabolomics; microbial; microbiome; microorganism; microorganism interaction; mouse model; mutant; mycobiome; novel therapeutics; pathogen; polymicrobial biofilm; preclinical study; response; transcriptome sequencing; Address; Adhesions; Affect; Anaerobic Bacteria; Anti-Bacterial Agents; Antifungal Agents; Bacteria; Candida; Candida albicans; Candida tropicalis; Chronic; Clinical Research; Coculture Techniques; Colitis; Colon; Communities; Crohn' s disease; Data; Diagnostic; Environment; Escherichia coli; Exhibits; Exposure to; Family; Feces; Filament; Flare; Gastrointestinal tract structure; Gene Expression; Generations; Genes; Goals; Habits; Hyphae; Immune response; In Vitro; Indoles; Infection; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Interruption; Intestinal Mucosa; Intestines; Knock-out; Lactobacillus; Letters; Lymphoid Cell; Mediating; Microbe; Microbial Biofilms; Modeling; Modification; Mucous Membrane; Mus; Oral; Organism; Outcome; Pathogenicity; Patients; Phase; Phenotype; Play; Prevalence; Process; Production; Pyrrolidonecarboxylic Acid; Recovery; Relapse; Reporting; Resolution; Role; Saccharomyces; Sampling; Serratia marcescens; Severities; Sodium Dextran Sulfate; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Symptoms; Testing; Therapeutic; Thick; Tissues; Trichosporon; Tryptophan; Ulcerative Colitis; Virulence Factors; Yeasts; aryl hydrocarbon receptor ligand; bacteriome; design; dextran sulfate sodium induced colitis; experimental study; fungus; gut microbiota; healing; host microbiota; in vivo; in vivo evaluation; in vivo imaging; indoleacetic acid; inflammatory disease of the intestine; inhibitor/antagonist; insight; interleukin-22; intestinal injury; metabolomics; microbial; microbiome; microorganism; microorganism interaction; mouse model; mutant; mycobiome; novel therapeutics; pathogen; polymicrobial biofilm; preclinical study; response; transcriptome sequencing

The role of gut fungal community (“mycobiome”, MYC) and interactions between the bacteriome (BM) and fungal communities in Crohn’s Disease (CD) have been relatively ignored. Recently, we compared the gut BM and MYC of CD patient’s to their healthy relatives and showed that abundance of the fungus Candida tropicalis (CT) was positively correlated with the bacteria Serratia marcescens (SM) and Escherichia coli (EC). We analyzed the number of CD patients that had all 3 species; the number CD patients exhibiting all 3 organisms was 30%, compared to 9% in healthy controls. Critically, abundance of all three organisms is 6.2 fold higher in CD patients vs. controls. Thus, the prevalence of this combination of microorganisms in the GI tracts of CD patients is significant. We further showed that ex-vivo, these polymicrobial triple species cooperated to form robust biofilms (TSBs) that were significantly increased compared to those formed by single-species (SSBs) or double-species (CT+EC or CT+SM, DSBs). Supporting data showed the interaction between CT, SM, and EC was specific; substitution of Trichosporon spp. (TC) or Saccharomyces fibuligera (SF) did not increase biofilm formation. However, preliminary data using Candida albicans (CA) as a control comparator reported by others to be elevated in CD patients, did cause increased TSB biofilm formation, indicating that this is a Candida specific effect. Importantly, we validated that the 3 pathogens formed robust biofilms in vivo (an anaerobic environment) using a dextran sodium sulfate (DSS) model of murine ulcerative colitis. Metabolomic analyses of supernatants from TSBs identified 11 significantly increased metabolites compared to SSBs or DSBs. Of these, 5-oxoproline (5-OP), was the most elevated metabolite (131-fold higher in TSBs compared to SSBs or DSBs). Also, Indole-3-acetic acid (IAA), formed as a result of Lactobacilli metabolizing tryptophan, which can act as an endogenous ligand for aryl hydrocarbon receptors (AhR), mediating innate lymphoid cell (ILC) production of IL-22, that in-turn triggers the inhibition of CA colonization, was also significantly increased. These 2 metabolites influence candidal virulence factors (filamentation, adhesion and auto-aggregation). Although these data are significant and indicate a strong polymicrobial interaction in CD, they do not establish the underlying mechanisms of these microbial interactions. This proposal focuses on determining the mechanism/s underlying the interaction of mixed species microbes using complementary in vitro and in vivo approaches. Our hypothesis is that SM, EC and Candida (CT and CA), in the setting of biofilms, interact cooperatively to exacerbate intestinal inflammation and exacerbate symptoms To address the mechanism(s) we will: (1) Determine the mechanism(s) metabolites formed by CT, EC, and SM in TSBs alter intestinal outcomes; (2) Identify gene changes underlying Candida modifications that increase pathogenicity using microbial mutant strains and RNAseq approaches; and (3) Determine whether exposure of TSBs to antibacterial, antifungal or metabolite inhibitors modulates the severity of intestinal inflammation in vivo. via multispecies interactions.

肠道真菌社区（“ Mycobiome”，Myc）的作用以及细菌（BM）与真菌之间的相互作用 克罗恩病（CD）的社区被相对忽略。最近，我们比较了肠道BM和MYC CD患者的健康亲属，并表明真菌念珠菌热带（CT）的抽象为 与细菌泥沙生（SM）和大肠杆菌（EC）呈正相关。 我们分析了 拥有所有3种物种的CD患者数量；表现出所有3个生物的数量CD患者为30％， 相比之下，健康对照组为9％。至关重要的是，CD患者的所有三种生物的抽象均高6.2倍 与控件。这是，这种微生物在CD患者的胃肠道中的这种组合的普遍性是显着的。 我们进一步表明，这些多菌三种物种是在体外，形成了强大的生物膜（TSB） 与由单物种（SSB）或双种物种（CT+EC或 CT+SM，DSB）。支持数据显示CT，SM和EC之间的相互作用是特定的。替换 Trichosporon spp。 （TC）或saccharomyces fibuligera（SF）不会增加生物膜的形成。但是，初步 使用白色念珠菌（CA）的数据作为对照比较者报告的对照比较剂，该比较剂将在CD患者中升高，DID 导致TSB生物膜形成增加，表明这是一种特异性效应。重要的是，我们进行了验证 3种病原体在体内形成了强大的生物膜（厌氧环境）使用葡萄糖钠 鼠溃疡性结肠炎的硫酸盐（DSS）模型。来自TSB的上清液的代谢组分析11 与SSB或DSB相比，代谢物显着增加。其中，5-氧化丙醇（5-OP）是最多的 代谢产物升高（与SSB或DSB相比，TSB中的131倍）。另外，形成的吲哚-3-乙酸（IAA） 由于乳酸杆菌代谢色氨酸，它可以充当芳基烃的内源配体 接收器（AHR），介导先天淋巴样细胞（ILC）的IL-22产生，这会触发CA的抑制 定殖，也显着增加。这两种代谢产物影响候选病毒因素 （细丝，粘合剂和自动聚集）。尽管这些数据很重要，并且表明很强 在CD中的多细胞相互作用，它们没有建立这些微生物相互作用的潜在机制。 该提案着重于确定使用混合物种微生物相互作用的机理 完成体外和体内方法。 我们的假设是SM，EC和念珠菌（CT和CA） 生物膜的设置，交互作用以加剧肠道注射并加剧症状 为了解决机制，我们将：（1）确定机制 TSB中由CT，EC和SM形成的代谢产物改变了肠结局； （2）确定基因变化 潜在的念珠菌修饰，使用微生物突变菌株和RNASEQ提高致病性 方法； （3）确定TSB是否暴露于抗菌，抗真菌或代谢物中 抑制剂调节体内肠炎的严重程度。 通过多人相互作用。