Discovery of Small Molecule Immunomodulators from Disease-Associated Microbiome Members

从疾病相关微生物组成员中发现小分子免疫调节剂

• 批准号: 9258585
• 负责人: David Richard Jackson
• 金额: $ 5.71万
• 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: 2017-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9258585
• 关键词: Affect; Anabolism; Anti-Inflammatory Agents; Anti-inflammatory; Area; Autistic Disorder; Autoimmune Diseases; Bacteria; Bioinformatics; Biological Assay; Biological Process; Birth; Bone Marrow; Cells; Chemicals; Clinical; Coupled; Data; Development; Diagnostic; Disease; Disease Progression; Environment; Enzymes; Evolution; Fractionation; Genes; Goals; Human; Human Microbiome; Immune response; Immune system; Immunology; Immunomodulators; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Institutes; Insulin-Dependent Diabetes Mellitus; Interleukin-10; Libraries; Link; Mammalian Cell; Metabolism; Microbe; Microbiology; Molecular; Molecular Bank; Monitor; Mus; Natural Products Chemistry; Onset of illness; Organism; Pathway interactions; Patient-Focused Outcomes; Patients; Physiological; Physiology; Plant Roots; Play; Prevention strategy; Process; Production; Regulation; Regulatory Element; Research; Resources; Role; Shapes; Statistical Data Interpretation; Structure; Systems Development; TNF gene; Testing; Therapeutic; Therapeutic Intervention; Training; Work; X ray diffraction analysis; X-Ray Diffraction; base; comparative genomics; cytokine; experimental study; genome analysis; gut microbiome; high throughput screening; host-microbe interactions; immunoregulation; in vivo; insight; knockout gene; macrophage; member; microbiome; prevent; skills; small molecule; transcription factor; transcriptome sequencing

Project Summary/Abstract The co-evolution of humans with bacteria has resulted in our bodies being colonized by numerous different species of microbes, collectively called the microbiome. Our intimate association with microbes, which begins even before birth, is especially important for immune system development. Changes in the gut microbiome have recently been linked to disease onset and progression for multiple autoimmune disorders – including type I diabetes (T1D), inflammatory bowel disease, and autism, but the molecular basis for how specific bacterial strains contribute to disease is unknown. The lack of mechanistic understanding of host-microbe interactions in disease prevents development of targeted microbiome-based therapies. The proposed research will identify bacterially produced molecules, and the mechanisms by which they promote or prevent autoimmune disease, with a particular focus on type I diabetes. First, I will generate a library of partially fractionated extracts from cultures of bacterial strains that were identified in a longitudinal T1D study. Strains will be selected based on their association with either disease onset and progression, or non-progression. The strains in this proposal are clinically derived and linked directly to patient outcomes. We predict that bacteria that promote T1D will produce inflammatory molecules, and bacteria that prevent T1D will produce anti-inflammatory molecules. We will test our molecular library using an IL-10/TNFα assay in murine bone marrow-derived macrophages (BMDMs), under both normal and inflammatory conditions. Fractions that either induce IL-10 or suppress TNFα under inflammatory conditions will be considered as anti-inflammatory leads; fractions that suppress IL-10 or induce TNFα will be inflammatory leads. We will conduct activity-guided fractionation to purify active immunomodulators, followed by structure determination using NMR, MS, and X-ray diffraction. Next, we will determine the biosynthetic pathways responsible for immunomodulators by sequencing and analyzing the genomes of the producers. This will enable manipulation of these genes to increase or decrease levels of immunomodulators in vivo. Determination of immunomodulator biosynthetic genes will allow for identification of similar genes in other organisms, and will drive discovery of related compounds with potentially increased or divergent bioactivity. Finally, we will determine immunomodulator modes of action in human cells using a two main functional assays: 1) cytokine profiling and 2) transcription factor-based RNA-seq (TF-seq). This work will expand our understanding of how the microbiome modulates host processes related to autoimmune disease, and provide a basis for therapeutic intervention.

项目总结/摘要 人类与细菌的共同进化导致我们的身体被许多不同的细菌所殖民。 微生物物种，统称为微生物组。我们与微生物的亲密关系， 甚至在出生前，对免疫系统的发育尤为重要。肠道微生物组的变化 最近与多种自身免疫性疾病的疾病发作和进展有关-包括 I型糖尿病（T1 D），炎症性肠病和自闭症，但如何具体细菌的分子基础 菌株对疾病的贡献是未知的。缺乏对宿主-微生物相互作用的机制理解， 疾病阻碍了基于微生物组的靶向治疗的发展。拟议的研究将确定 细菌产生的分子，以及它们促进或预防自身免疫性疾病的机制， 特别关注I型糖尿病。 首先，我将从细菌菌株的培养物中产生一个部分分级提取物的文库， 在T1 D纵向研究中发现。菌株将根据其与任一疾病的相关性进行选择 发作和进展或非进展。本提案中的菌株来自临床，并与 病人的结果。我们预测，促进T1 D的细菌会产生炎症分子， 预防T1 D的细菌会产生抗炎分子。我们将使用一种 在正常和非正常条件下， 炎性条件。在炎症条件下诱导IL-10或抑制TNFα的组分 将被视为抗炎先导物;抑制IL-10或诱导TNFα的组分将被 煽动性的线索我们将进行活性导向分级分离以纯化活性免疫调节剂， 通过NMR、MS和X射线衍射进行结构测定。 接下来，我们将通过测序和测序来确定负责免疫调节剂的生物合成途径。 分析生产者的基因组。这将使操纵这些基因，以增加或减少 体内免疫调节剂水平。免疫调节剂生物合成基因的确定将允许 在其他生物体中识别类似基因，并将推动发现相关化合物， 增加或发散的生物活性。最后，我们将确定免疫调节剂在人体细胞中的作用模式 使用两种主要的功能测定：1）细胞因子谱分析和2）基于转录因子的RNA-seq（TF-seq）。 这项工作将扩大我们对微生物组如何调节与以下相关的宿主过程的理解： 自身免疫性疾病，并提供治疗干预的基础。