A probiotic-derived protein regulates epigenetic programming in intestinal epithelial cells for long-term prevention of colitis

益生菌衍生的蛋白质调节肠上皮细胞的表观遗传编程，以长期预防结肠炎

• 批准号: 10386935
• 负责人: FANG YAN
• 金额: $ 44.15万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10386935
• 关键词: Address; Adult; Bacteria; Binding; CD4 Positive T Lymphocytes; Cell Line; Cell division; Cell physiology; Child; Chromatin; Clinical Trials; Colitis; Complex; Data; Deposition; Dimerization; Early Intervention; Ecosystem; Enhancers; Epidermal Growth Factor Receptor; Epigenetic Process; Epithelial; Epithelial Cells; Exposure to; Foundations; Funding; Gene Expression; Gene Proteins; Genes; Health; Health Promotion; Histone H3; Histones; Human; Incidence; Individual; Infant; Inflammation; Inflammatory Bowel Diseases; Inherited; Intervention; Intestines; Lactobacillus casei rhamnosus; Lamina Propria; Life; Longevity; Lysine; Mediating; Methyltransferase; Modeling; Mus; Natural regeneration; Neonatal; Outcome; Predisposition; Prevention; Prevention strategy; Prevention therapy; Probiotics; Production; Proteins; Public Health; Regulatory T-Lymphocyte; Reporting; Research; Research Support; Signal Transduction; Supplementation; Testing; Tight Junctions; Transactivation; Transcriptional Activation; Transforming Growth Factor beta; Translational Research; Up-Regulation; Work; antibody inhibitor; autoinflammatory; dimer; dysbiosis; epigenetic memory; gut inflammation; gut microbiome; gut microbiota; high risk; histone methyltransferase; histone modification; imprint; intestinal epithelium; intestinal homeostasis; knock-down; microbiome; mouse model; neonatal period; neonate; neutralizing antibody; novel; novel therapeutic intervention; novel therapeutics; postnatal development; preservation; prevent; programs; promoter; receptor; response; secretory protein; stem cells; transcription factor

PROJECT SUMMARY Dysbiosis in infants and children is associated with increased susceptibility to inflammatory bowel disease (IBD) in adults. However, the mechanisms whereby the gut microbiota colonization in early life confers health outcomes throughout the lifespan remain unclear. Studies from the previous funding period isolated and cloned a Lactobacillus rhamnosus GG (LGG)-derived secretory protein, p40, and demonstrated that neonatal p40 supplementation prevents colitis in adult mice. Our preliminary studies discovered that p40 interacts with two transcriptional factors, Mga and Max, to regulate expression of Setd1β gene, which encodes a methyltransferase for catalyzing mono and trimethylation of histone 3-lysine 4 (H3K4me1/3). We identified TGFβ as a potential target of p40-up-regulated Setd1β. IECs are rapidly renewed and continuously regenerated from intestinal stem cells (ISCs). We found that p40 modulates H3K4m31/3 in ISCs in early stage, and p40 treatment in neonates, but not adult mice, stimulates sustained increase in TGFβ gene expression in IECs. TGFβ has been shown to have multiple functions against inflammation. Thus, we hypothesize that p40 up-regulates Setd1β gene expression in IECs through increasing Mga:Max dimerization. Supplementation with p40 in early life stimulates Setd1β-dependent H3K4me1/3 deposition at the TGFβ locus in ISCs, which is inherited by IECs to enable the sustained increase in TGFβ production, and subsequently prevention of intestinal inflammation in adulthood. In Aim 1, we will determine whether p40-stimulated Setd1β gene expression and H3K4me1/3 are required to drive the increase in expressing TGFβ gene in IECs, and elucidate whether the interaction between p40 and the Mga:Max dimers mediates up-regulation of Setd1β production. In Aim 2, we will identify the temporal window of p40 exposure in early life that causes the sustained increase in TGFβ production in IECs, and define whether p40-regulated sustained increase in TGFβ gene expression in IECs requires the increase of Setd1β gene expression inISCs in early life. Human and mouse enteroids and colonoids and 2D cultures and cell lines with silencing Setd1β or Mga genes, and mouse models of constitutive and inducible Setd1β gene knock down in IECs or ISCs will be utilized for these two aims. In Aim 3, we will determine whether the sustained increase in TGFβ production after p40 supplementation in early life is required to prevent colitis in adult mice. We will use neutralizing antibodies and inhibitors to block TGFβ function in mice with induced and spontaneously developed colitis. We will also determine whether sustained increase in TGFβ production enhances Tregs induction in the intestine and protective epithelial responses for the prevention of colitis in adulthood. This aim will be tested in mice with TGFβ receptor II deletion in CD4+ T cells and Smad4 deletion in IECs. Together, our proposed research will elucidate a novel mechanism underlying the consequences of long-lasting effects of p40 on prevention of colitis, and lay the foundation for developing early intervention with p40 as a novel therapy for prevention of IBD in adults.

项目总结