Regulation of intestinal development by a Lactobacillus GG-derived protein

乳杆菌 GG 衍生蛋白对肠道发育的调节

• 批准号: 8964588
• 负责人: FANG YAN
• 金额: $ 39.25万
• 依托单位: VANDERBILT UNIVERSITY
• 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 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8964588
• 关键词: Address; Adult; Apoptosis; Biological Assay; Biological Preservation; Cell Line; Cell Proliferation; Child; Clinical; Colitis; Colon; DTR gene; Data; Development; Disease; Disintegrins; Epidermal Growth Factor Receptor; Epithelial; Epithelial Cell Proliferation; Epithelial Cells; Gastrointestinal tract structure; Gene Expression Profile; Genes; Growth; Health; Human; Hydrogels; Immune; Immune system; Immunocompromised Host; Individual; Inflammation; Injury; Interleukin-9; Interruption; Intestines; Knowledge; Lactobacillus; Lactobacillus casei rhamnosus; Life; Ligands; Mediating; Metalloproteases; Mucins; Mus; Nature; Neonatal; Nutrient; Nutritional; Organ Culture Techniques; Pectins; Persons; Physiological; Physiological Processes; Play; Predisposition; Probiotics; Production; Proteins; Publishing; Receptor Activation; Receptor Signaling; Regulation; Research; Role; Signal Pathway; Signal Transduction; Small Interfering RNA; Staging; System; Testing; Tight Junctions; Tissues; Transcriptional Regulation; Up-Regulation; Villus; Weaning; Weight Gain; Work; Zein; base; clinical application; clinical efficacy; cytokine; disorder prevention; fetal; gut microbiota; intestinal epithelium; migration; mouse model; nutrient metabolism; organ growth; postnatal; prevent; public health relevance; research study; response; tissue culture; uptake; Address; Adult; Apoptosis; Biological Assay; Biological Preservation; Cell Line; Cell Proliferation; Child; Clinical; Colitis; Colon; DTR gene; Data; Development; Disease; Disintegrins; Epidermal Growth Factor Receptor; Epithelial; Epithelial Cell Proliferation; Epithelial Cells; Gastrointestinal tract structure; Gene Expression Profile; Genes; Growth; Health; Human; Hydrogels; Immune; Immune system; Immunocompromised Host; Individual; Inflammation; Injury; Interleukin-9; Interruption; Intestines; Knowledge; Lactobacillus; Lactobacillus casei rhamnosus; Life; Ligands; Mediating; Metalloproteases; Mucins; Mus; Nature; Neonatal; Nutrient; Nutritional; Organ Culture Techniques; Pectins; Persons; Physiological; Physiological Processes; Play; Predisposition; Probiotics; Production; Proteins; Publishing; Receptor Activation; Receptor Signaling; Regulation; Research; Role; Signal Pathway; Signal Transduction; Small Interfering RNA; Staging; System; Testing; Tight Junctions; Tissues; Transcriptional Regulation; Up-Regulation; Villus; Weaning; Weight Gain; Work; Zein; base; clinical application; clinical efficacy; cytokine; disorder prevention; fetal; gut microbiota; intestinal epithelium; migration; mouse model; nutrient metabolism; organ growth; postnatal; prevent; public health relevance; research study; response; tissue culture; uptake

 DESCRIPTION (provided by applicant): The symbiotic relationship between intestinal microbiota and the host plays an important role in the intestinal development. Interruption of this relationship leads to infectious and immune-mediated diseases. This underscores the importance of identifying the effects of probiotics on intestinal development and disease prevention in children. However, there are critical gaps in knowledge about mechanisms of probiotic action. The efficacy of probiotics for nutritional and clinical application remains to be convincingly established. p40, a LGG-derived protein isolated and cloned by our group, activates epidermal growth factor receptor (EGFR) and its downstream target, PI3K/Akt in intestinal epithelial cells through stimulation of EGFR ligand release. Activation of EGFR by p40 is required for inhibition of cytokine-induced apoptosis, preservation of barrier function, and enhancement of mucin production in intestinal epithelial cells. We have generated a pectin/zein hydrogel bead system to specifically delivered p40 to the colon in mouse. By using this delivery system p40 has been sown to prevent and treat experimental colitis in mice in an EGFR-dependent manner. Since activation of EGFR plays a physiological role in development, we have expanded the scope of our work to investigate the effects of p40 on intestinal development. Based on our preliminary results that LGG colonization promotes functional maturation of the intestine and decreases susceptibility to intestinal injury and colitis in adult mice, three integrated specific aims are proposed to enhance knowledge of the functional nature of p40 in regulation of intestinal development and mechanisms of the action of p40. In Specific Aim 1, the roles of p40 in intestinal functional maturation will be determined. We will use neonatal mice, human fetal intestinal organ culture, and human and mouse enteroids to determine the roles of p40 in intestinal epithelial cell proliferation, differentiation, migration,and digestive and absorptive capacities and epithelial tight junction formation. In Specific Aim 2, transcriptional targets in intestinal epithelial cells involved in p40-regulated cellular responsesto promote intestinal functional maturation will be defined. We will determine p40-regulated intestinal epithelial genes in neonatal mice and in human fetal intestinal organ culture, and verif the function of individual candidate p40-regulated genes predicted to regulate epithelial proliferation, differentiation, migration, transporters, and tight junction formation using in vitr and ex vivo assays. In Specific Aim 3, the p40- stimulated signaling pathway in intestinal epithelial cells that is required for promotion of intestinal functional maturation will be elucidaed. We will use mouse models with intestinal epithelial cell-specific deficiency of EGFR and PI3K and pharmacologic inhibition of EGFR and Akt in human fetal intestinal organ culture and enteroids to determine the requirement of EGFR and Akt activation for p40-stimulated epithelial responses and p40-regulated EGFR and Akt-dependent transcriptional targets for functional maturation. These studies will contribute new information for nutritional and clinical applications of p40 for health in children.

 描述（由适用提供）：肠道菌群与宿主之间的共生关系在肠发展中起着重要作用。中断 关系导致传染性和免疫介导的疾病。这强调了确定益生菌对儿童肠发展和疾病预防的影响的重要性。但是，关于益生菌作用机制的知识存在关键的差距。益生菌对营养和临床应用的有效性尚待 令人信服地建立了。 P40是由我们组分离和克隆的LGG衍生蛋白，通过刺激EGFR配体释放，激活表皮生长因子受体（EGFR）及其下游靶标的PI3K/AKT。抑制细胞因子诱导的细胞凋亡，屏障功能的保存以及肠上皮细胞中粘蛋白产生的增强需要p40激活EGFR。我们已经生成了果胶/锌水凝胶珠系统，以将P40专门传递到小鼠中的结肠。通过使用此输送系统，P40已被播种，以预防和治疗小鼠以EGFR依赖的方式治疗实验性结肠炎。由于EGFR的激活在发育中起着身体的作用，因此我们扩大了工作范围，以研究p40对肠道发育的影响。基于我们的初步结果，LGG定殖促进了肠道的功能成熟，并降低了成年小鼠肠道损伤和结肠炎的易感性，提出了三个综合特定目的，以增强对P40在肠道发展和P40作用机制的调节中的功能性质的了解。在特定的目标1中，将确定p40肠功能成熟的作用。我们将使用新生儿小鼠，人类胎儿肠道器官培养物以及人和小鼠肠to虫来确定p40肠上皮细胞增殖，分化，迁移，消化和吸收性能力以及上皮紧密连接形成的作用。在特定的目标2中，将定义参与P40调节的细胞反应的肠上皮细胞中的转录靶标。我们将确定新生小鼠和人胎儿肠道器官培养的p40调节的肠上皮基因，并通过使用In Vitr和ex Vivo Assays进行调节的单个候选p40调节基因的功能，并验证其p4候选p40调节的基因的功能。在特定的目标3中，将要促进肠上皮细胞中的P40刺激的信号通路，以促进肠功能成熟。我们将使用具有EGFR和PI3K的肠上皮细胞特异性缺乏的小鼠模型，以及对EGFR和AKT的药物抑制在人类胎儿肠道器官培养物和肠to型中，以确定EGFR和AKT激活对P40刺激的EGFR和AKT激活的需求。这些研究将为营养和临床应用提供新信息 儿童健康的P40。