Defining bioactivities of peptides released from human milk proteins in the preterm infant intestine

定义早产儿肠道中母乳蛋白释放的肽的生物活性

• 批准号: 10658669
• 负责人: Brian Scottoline
• 金额: $ 65.8万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-15 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10658669
• 关键词: Address; Amino Acids; Anti-Bacterial Agents; Biological; Biological Assay; Biological Models; Biology; Brain Injuries; Bronchopulmonary Dysplasia; Cattle; Cell Proliferation; Cell physiology; Cellular biology; Childhood; Data; Databases; Development; Digestion; Enteral Nutrition; Epithelial Cells; Exhibits; Gastrointestinal tract structure; Gene Expression; Goals; Growth; Growth and Development function; Health Promotion; Human; Human Biology; Human Milk; Immune system; In Vitro; Infant; Infant Health; Inflammatory; Intestinal Content; Intestines; Liquid substance; Macrophage; Mass Spectrum Analysis; Measures; Mediating; Milk; Milk Proteins; Minimum Inhibitory Concentration measurement; Mission; Modeling; National Institute of Child Health and Human Development; Necrotizing Enterocolitis; Neonatal Intensive Care Units; Neonatology; Peptide Synthesis; Peptides; Physiology; Premature Infant; Proliferating; Proteins; Proteomics; Public Health; R peptide; Research; Research Priority; Retinopathy of Prematurity; Risk; Role; Sampling; Source; Stimulus; Techniques; Testing; Time; Work; antimicrobial; candidate identification; cytokine; feeding; fortification; gut health; gut microbiome; immunoregulation; improved; in vitro activity; in vitro testing; in vivo; innovation; intestinal epithelium; late onset sepsis; microbiome; neonatal care; neonatal human; novel; nutrition; pasteurization; preterm newborn; protein expression; response; sample collection; statistics

PROJECT SUMMARY Human milk, whether maternal or pasteurized donor human milk, is considered the ideal nutrition for preterm infants. In addition to serving as a source of amino acids to support infant growth, human milk proteins encode cryptic bioactive peptides released during infant digestion. These peptides can be found in the infant intestine luminal fluid and have an array of biological activities, including antimicrobial, bifidogenic, immunomodulatory, and effects on intestinal physiology. Although peptides responsible peptides our initial work demonstrates that milk released and present in the infant intestine have beneficial activities, identifying the specific peptides for these gut-related actions has been limited. Thus, there is a critical need to determine the specific released within the infant's gastrointestinal tract that mediate these bioactivities. Our long-term goal is to leverage the biology of human milk to promote optimal development and growth of preterm infants ex utero. The overall objective of this proposal is to demonstrate the presence and identities of gut health-promoting human milk protein-derived peptides within the preterm infant intestine. Our central hypothesis is that bioactive peptides are released in the infant intestine to modulate gut physiology through the microbiome, intestinal epithelium, and immune system. Our specific aims are to determine the 1) antimicrobial and bifidogenic activity, 2) intestinal epithelial cell (IEC)-specific effects (barrier function, proliferation, differentiation) and 3) macrophage-immunomodulatory activity of peptides in the intestinal contents of preterm infants fed human milk. For each of these activities, we will identify and validate specific peptides. Our approach will be to collect intestinal samples from preterm neonates in the neonatal intensive care unit (NICU) after feeding human milk, extract the peptide component from these samples, test their antibacterial, bifidogenic, IEC-modulatory and macrophage-immunomodulatory activity in vitro, and identify candidate bioactive peptides via mass spectrometry, database searching, and statistical techniques to be synthesized and tested for function. As a sub-aim, we will perform this analysis for maternal and pasteurized donor human milk. This research is innovative because it identifies, for the first time, bioactivities of peptides released during in vivo infant digestion, provides a means for identifying candidate bioactive peptides and establishes a pipeline for testing candidate peptides in relevant model systems, including neonatal human enteroids. At the conclusion of this project, we expect to 1) identify the gut health-related bioactivity of human milk protein-derived peptide extracts from the preterm infant intestine and 2) identify specific peptides present in the infant intestine that can modulate the microbiome, IECs, and macrophages. The positive impact of this research is that it will provide the basis for leveraging milk-derived, bioactive peptides to improve infant gut health for preterm and potentially all infants.

项目概要 母乳，无论是母乳还是经过巴氏灭菌的捐赠母乳，都被认为是婴儿的理想营养品。 早产儿。除了作为支持婴儿生长的氨基酸来源外，母乳蛋白 编码婴儿消化过程中释放的神秘生物活性肽。这些肽可以在婴儿体内找到 肠腔液并具有一系列生物活性，包括抗菌、双歧杆菌、 免疫调节和对肠道生理的影响。虽然 肽 负责任的 肽 我们的初步工作表明牛奶 释放并存在于婴儿肠道中具有有益的活性，可识别特定的肽 因为这些与肠道相关的行动受到了限制。因此，迫切需要确定具体的 在婴儿胃肠道内释放，介导这些生物活性。 我们的长期目标是利用母乳的生物学特性促进最佳发育和生长 宫外早产儿。该提案的总体目标是展示存在和身份 早产儿肠道内促进肠道健康的母乳蛋白衍生肽。我们的中央 假设是婴儿肠道中释放生物活性肽，通过 微生物组、肠上皮和免疫系统。我们的具体目标是确定 1) 抗菌剂 和双歧活性，2) 肠上皮细胞 (IEC) 特异性效应（屏障功能、增殖、 分化）和3）早产儿肠道内容物中肽的巨噬细胞免疫调节活性 婴儿喂养母乳。对于每项活动，我们将识别并验证特定的肽。我们的方法 将在新生儿重症监护室（NICU）收集早产儿的肠道样本 喂养母乳，从这些样品中提取肽成分，测试其抗菌、双歧杆菌、 体外 IEC 调节和巨噬细胞免疫调节活性，并鉴定候选生物活性肽 通过质谱、数据库搜索和统计技术进行合成和功能测试。 作为一个子目标，我们将对母乳和经过巴氏灭菌的捐赠母乳进行分析。 这项研究具有创新性，因为它首次鉴定了在过程中释放的肽的生物活性。 婴儿体内消化，提供了识别候选生物活性肽的方法并建立了管道 用于测试相关模型系统中的候选肽，包括新生儿人肠类。结论时 在这个项目中，我们期望 1) 确定母乳蛋白衍生肽与肠道健康相关的生物活性 早产儿肠道提取物，2) 鉴定婴儿肠道中存在的特定肽，这些肽可以 调节微生物组、IEC 和巨噬细胞。这项研究的积极影响在于它将提供 利用乳源生物活性肽改善早产儿和潜在婴儿肠道健康的基础 所有婴儿。