Role of innate immunity and the microbiome in colitis-associated dysplasia

先天免疫和微生物组在结肠炎相关发育不良中的作用

基本信息

批准号：
10659444
负责人：
Maria Teresa Abreu
金额：
$ 38.94万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2027-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10659444
关键词：
Animal Model Bacteria Bacterial Translocation Biopsy Cells Characteristics Chronic Colectomy Colitis Colon Colon Carcinoma Colonic inflammation Cytometry DNA Damage Data Defect Dependence Development Dysplasia Epithelial Cells Epithelium Feces Fright Functional disorder Funding Generations Genetic Transcription Germ-Free Human Hydrogen Peroxide Image Immune signaling Immune system Inflammation Inflammatory Intestines Lesion Link Liver Mediating Metagenomics Methods Modeling Mucous Membrane Mus Myeloid-derived suppressor cells NADPH Oxidase Natural Immunity Oxidases Oxidative Stress Oxygen Pathogenesis Pathway interactions Patients Permeability Predisposition Production Proteobacteria Research Personnel Risk Role Sampling Signal Induction Signal Transduction Specimen T-Lymphocyte TLR4 gene Techniques Testing Translating Tumor Promotion Ulcerative Colitis Work biobank chemokine colitis associated cancer colitis-associated neoplasia colon dysplasia cytokine deep sequencing dysbiosis gut bacteria intestinal epithelium metabolome metabolomics microbial microbiome monolayer mouse model premalignant prevent recruit single-cell RNA sequencing translational approach tumor tumor microbiome tumorigenesis tumorigenic villin

项目摘要

PROJECT ABSTRACT The two greatest fears expressed by patients with ulcerative colitis (UC) are developing colon cancer and losing their colon. Although colitis-associated cancer is less common, dysplasia is more common and often results in colectomy. Our proposal leverages the work of the previous funding period and advances in the field to take a translational approach to unraveling dysplasia in colitis. The focus of our studies has been the link between innate immune signaling and intestinal bacteria leading to colitis-associated neoplasia. Our results have led us to focus on the role of dual oxidase 2 (DUOX2) in dysplasia. DUOX2 is a NADPH oxidase that catalyzes the conversion of oxygen into hydrogen peroxide (H2O2) upon interaction with the maturation factor DUOXA2. It is consistently upregulated in biopsies from IBD patients and DUOX2/DUOXA2 expression is further increased in patients who have had dysplasia. Our group has shown that both inflammatory and microbial signals induce the expression and activity (H2O2 production) of DUOX2 in colonic epithelial cells. We have proven that chronic activation of DUOX2 leads to the formation of tumors, which is almost totally abrogated by inactivating DUOX2 in the epithelium. In the current proposal, we hypothesize that IBD-associated dysbiosis activates Duox2 and local production of H2O2 leading to epithelial barrier dysfunction, recruitment of tumor-promoting myeloid derived suppressor cells (MDSCs), and generation of a tumorigenic microbiome in a feed forward loop. This is pursued in the following specific aims: 1) Determine the dependence of inflammatory colonic dysplasia on epithelial Duox2 signaling. Here we will investigate epithelial barrier dysfunction and DNA damage pathways caused by epithelial Duox2 activation using colonoid models (human and murine). Using humanized germ-free mice, we will determine if the UC-dysplasia microbiome is sufficient to cause DUOX2-mediated permeability defects and DNA damage. 2) Dissect the role of tumor-promoting MDSCs on dysplasia development in the setting of DUOX2-mediated oxidative stress. Our preliminary data demonstrate that TLR4-driven tumors have enhanced recruitment of tumor supporting MDSCs. Here we will identify DUOX2-dependent epithelial factors (chemokines/cytokines) and the transcriptional (single cell-RNA seq) and functional (T cell suppressive) characteristics of MDSCs in our murine models of tumorigenesis and UC patients with dysplasia. 3) Identify targetable microbial pathways linked to Duox2 activation and dysplasia. Transfer of the microbiome from tumor-susceptible villin-TLR4 mice is sufficient to transfer CAC susceptibility. New metagenomic and metabolomic data show clear differences between UC dysplasia and UC without dysplasia. We will use our mouse models and UC dysplasia samples to perform deep sequencing and metabolomic characterization of the tumor-promoting microbiome. Metabolites will be tested for their ability to induce or inhibit H2O2/DUOX2 in colonoids. The work proposed herein will provide the mechanistic justification for subsequent human studies to target Duox2 and specific microbial pathways to halt progression from UC inflammation to dysplasia.

项目摘要溃疡性结肠炎（UC）患者表达的两个最大的恐惧正在发生结肠癌并失去他们的结肠。尽管与结肠炎相关的癌症不太常见，但发育异常更为普遍，通常会导致结肠切除术。我们的提议利用了上一期资金期的工作，并在该领域的进步转化方法是结肠炎中发育不良的方法。我们研究的重点是先天免疫信号传导和肠道细菌，导致结肠炎相关的肿瘤。我们的结果带领我们专注于双重氧化酶2（DUOX2）在发育不良中的作用。 DUOX2是一种NADPH氧化酶，可催化与成熟因子duoxa2相互作用后，氧氧转化为过氧化氢（H2O2）。这是来自IBD患者的活检中始终上调，DUOX2/DUOXA2表达进一步增加患有发育不良的患者。我们的小组表明，炎症和微生物信号都诱导 DUOX2在结肠上皮细胞中的表达和活性（H2O2产生）。我们已经证明了慢性 DUOX2的激活导致肿瘤的形成，这几乎通过灭活DUOX2完全消除在上皮中。在当前的提案中，我们假设与IBD相关的营养不良激活DUOX2和 H2O2的局部生产导致上皮屏障功能障碍，促进肿瘤髓样的募集抑制细胞（MDSC），并在饲料前循环中产生肿瘤的微生物组。这是追求的在以下特定目的中：1）确定炎症性结肠发育不良的依赖性对上皮 DUOX2信号传导。在这里，我们将研究由使用结肠模型（人和鼠）激活上皮DUOX2。使用无源细菌的小鼠，我们将确定UC-DYPLASIA微生物组是否足以引起DUOX2介导的渗透性缺陷和 DNA损伤。 2）剖析促进肿瘤的MDSC在发育不良发育中的作用 DUOX2介导的氧化应激。我们的初步数据表明，TLR4驱动的肿瘤已增强募集支持MDSC的肿瘤。在这里，我们将确定依赖DUOX2的上皮因素（趋化因子/细胞因子）和转录（单细胞-RNA SEQ）和功能（T细胞抑制） MDSC的特征在我们的鼠类肿瘤发生模型和UC发育不良患者的特征中。 3）识别与DUOX2激活和发育不良有关的可靶向微生物途径。从微生物组转移肿瘤敏感的Villin-TLR4小鼠足以传递CAC敏感性。新的宏基因组和代谢组数据显示，没有发育异常的UC发育不良和UC之间存在明显的差异。我们将使用我们的小鼠模型和UC发育不良样品，以执行深层测序和代谢组合表征促进肿瘤的微生物组。代谢物将测试其诱导或抑制H2O2/DUOX2的能力结石。本文提出的工作将为后续研究提供机械辩护靶向DUOX2和特定的微生物途径，以阻止从UC炎症到发育不良的进展。

项目成果

期刊论文数量（37）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Practical techniques for detection of Toll-like receptor-4 in the human intestine.

检测人肠道中 Toll 样受体 4 的实用技术。

DOI：
10.1007/978-1-59745-541-1_21
发表时间：
2009
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
0
作者：
Ungaro,Ryan;Abreu,MariaT;Fukata,Masayuki
通讯作者：
Fukata,Masayuki

Intestinal Epithelial Inactivity of Dual Oxidase 2 Results in Microbiome-Mediated Metabolic Syndrome.

DOI：
10.1016/j.jcmgh.2023.06.009
发表时间：
2023
期刊：
CELLULAR AND MOLECULAR GASTROENTEROLOGY AND HEPATOLOGY
影响因子：
7.2
作者：
Hazime, Hajar;Ducasa, G. Michelle;Santander, Ana M.;Brito, Nivis;Gonzalez, Eddy E.;Ban, Yuguang;Kaunitz, Jonathan;Akiba, Yasutada;Fernandez, Irina;Burgueno, Juan F.;Abreu, Maria T.
通讯作者：
Abreu, Maria T.

Pathogen recognition receptors, cancer and inflammation in the gut.