Identification and characterization of gut microbial bioactive molecules that determine predisposition to autoimmune disease and atopy

确定自身免疫性疾病和特应性倾向的肠道微生物生物活性分子的鉴定和表征

• 批准号: 9889899
• 负责人: Jon Clardy
• 金额: $ 66.91万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9889899
• 关键词: Address; Affect; Allergic Disease; Anti-Inflammatory Agents; Appearance; Archives; Autoimmune Diabetes; Autoimmune Diseases; Bacteria; Bioinformatics; Biological; Biological Assay; Biology; Biopsy; Brain; Cell Extracts; Cell Line; Cells; Cellular Stress; Chemicals; Coculture Techniques; Complex; Coupled; Crohn' s disease; Crystallization; Culture Media; Culture Techniques; Data; Data Set; Dendritic Cells; Development; Digestion; Disease; Disease Progression; Dose; Environment; Epithelial Cells; Finland; Fractionation; Gastrointestinal Diseases; Gene Cluster; Genetic; Genetic Predisposition to Disease; Hand; Heart; High Pressure Liquid Chromatography; Human; Hypersensitivity; Image; Immune System Diseases; Immune response; Immune system; Immunity; Incidence; Individual; Infant; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Inflammatory Response; Infusion procedures; Insulin-Dependent Diabetes Mellitus; Intestines; Laboratories; Lead; Libraries; Link; Living Standards; Mass Spectrum Analysis; Measurement; Measures; Membrane; Metabolism; Methodology; Microbe; Modification; Molecular; Molecular Biology; Molecular Weight; Monitor; Mucous Membrane; Mucous body substance; Myeloid Cells; NIH Program Announcements; Neurotransmitters; Nuclear Magnetic Resonance; Organoids; Pathway interactions; Patients; Physiological; Play; Population; Porifera; Predisposition; Probiotics; Production; Public Health; Reporter; Role; Sampling; Signal Pathway; Signal Transduction; Signaling Molecule; Structure; System; TNF gene; Techniques; Therapeutic; Therapeutic Intervention; Ulcerative Colitis; Vitamins; Work; X ray diffraction analysis; aqueous; atopy; base; body system; cell type; cohort; crystallinity; design; detector; gut bacteria; gut microbiome; gut microbiota; immune activation; immunoregulation; insight; member; microbial; monolayer; novel; response; screening; small molecule; stem; stool sample; therapeutic target; three dimensional structure; transcription factor; two-dimensional

ABSTRACT We know that the gut microbiome functions as an accessory organ system that plays critical roles in human digestion; the production of essential vitamins, neurotransmitters, and other small molecules; and in the regulation of the immune system. We also know that changes in the gut microbiota can lead to disease, especially inflammatory bowel disease and related inflammatory disorders. However, we know very little about the molecules and mechanisms that connect gut microbiota to these diseases. As the bacteria in the gut microbiota sense and respond to their environment with small molecules, it's likely that some of these molecules are key regulators of both pro-inflammatory and anti-inflammatory responses. This project seeks to identify these molecules and the mechanisms by which they operate. The project stems from a rich data set (DIABIMMUNE) that revealed a number of robust correlations between the appearance, or disappearance, of members of the gut microbiota and the development of dysregulated immune responses and disease in infants with genetic pre-disposition to type 1 diabetes. We will focus on ~40 strains that appear to play outsize roles in disease progression to discover what pro- or anti-inflammatory small molecule signals they produce. In Specific Aim 1 we will culture patient-derived strains of these bacteria under a variety of conditions, capture the small molecules produced, and measure their ability to regulate a pro-inflammatory signal (TNFalpha) and an anti- inflammatory signal (IL-10). The most potent and selective regulators will advance to Specific Aim 2 in which bioassay-guided fractionation will guide the isolation of active molecules, which will then be structurally characterized with spectroscopic (both nuclear magnetic resonance and mass spectrometry) and imaging (X-ray diffraction) techniques. With known molecules linked to confirmed biological activity in hand, we will prioritize hits based on their presence in the DIABIMMUNE stool samples, which we have archived. Only bioactive molecules present in human samples will be analyzed in specific Aim 3 to determine mechanism. Because the signaling pathways through which these molecules exert their biological effects are complex, we will employ a specially designed and newly implemented assay that simultaneously measures more than 50 transcription factors that regulate inflammation, immunity, metabolism and cell stress. By profiling transcription factor activity and comparing to known compounds, we will be able to classify molecules based on their activity profiles. We will further characterize their mechanism of action using a variety of functional assays on both patient derived myeloid cells as well as colonic and ileal mucosa cell lines that develop into four distinct cell types that form three-dimensional structures including crypts covered by a secreted mucus layer. At the end of the pipeline formed by these three specific aims, small molecules that regulate immune responses along annotated pathways will be known, and this identification of both molecule and mechanism will provide insights into both basic biology and potential therapeutic interventions.

抽象的 我们知道，肠道微生物组作为辅助器官系统，在人类健康中发挥着关键作用。 消化;生产必需的维生素、神经递质和其他小分子；并在 免疫系统的调节。我们还知道肠道微生物群的变化会导致疾病， 特别是炎症性肠病和相关的炎症性疾病。然而我们对此知之甚少 将肠道微生物群与这些疾病联系起来的分子和机制。由于肠道内的细菌 微生物群通过小分子感知环境并做出反应，其中一些分子很可能 是促炎和抗炎反应的关键调节因子。该项目旨在确定 这些分子及其运作机制。该项目源于丰富的数据集 （DIABIMMUNE）揭示了许多存在或消失之间的强大相关性 肠道微生物群成员以及婴儿免疫反应失调和疾病的发展 具有 1 型糖尿病的遗传倾向。我们将重点关注约 40 种似乎在 疾病进展，以发现它们产生什么促炎或抗炎小分子信号。具体来说 目标 1 我们将在各种条件下培养源自患者的这些细菌菌株，捕获小的 产生的分子，并测量它们调节促炎信号（TNFα）和抗炎信号的能力 炎症信号（IL-10）。最有效和最有选择性的监管机构将推进到具体目标 2，其中 生物测定引导的分级分离将指导活性分子的分离，然后在结构上进行分离 通过光谱（核磁共振和质谱）和成像（X 射线）表征 衍射）技术。有了与手头已确认的生物活性相关的已知分子，我们将优先考虑 根据我们已存档的 DIABIMMUNE 粪便样本中的存在情况进行命中。仅有生物活性 人类样本中存在的分子将在特定目标 3 中进行分析以确定机制。因为 这些分子发挥其生物效应的信号传导途径很复杂，我们将采用 专门设计和新实施的检测方法可同时测量 50 多个转录 调节炎症、免疫、新陈代谢和细胞应激的因素。通过分析转录因子活性 与已知化合物相比，我们将能够根据分子的活性特征对分子进行分类。我们 将使用对两种患者来源的多种功能测定进一步表征其作用机制 骨髓细胞以及结肠和回肠粘膜细胞系发育成四种不同的细胞类型 三维结构，包括被分泌的粘液层覆盖的隐窝。在管道的末端 由这三个特定目标形成，沿着注释调节免疫反应的小分子 途径将是已知的，对分子和机制的识别将提供对两者的见解 基础生物学和潜在的治疗干预措施。