Evaluation of recombinant bovine lactoferrin and N-linked glycan effects on infant gut health and immunity

重组牛乳铁蛋白和 N 连接聚糖对婴儿肠道健康和免疫力影响的评估

• 批准号: 10821138
• 负责人: Vanessa Patricia Dunne-Castagna
• 金额: $ 31.13万
• 依托单位: TURTLETREE LABS INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-19 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10821138
• 关键词: Address; Agriculture; Animals; Anti-Inflammatory Agents; Antigens; Back; Binding; Biological; Breast Feeding; Breastfed infant; CD4 Positive T Lymphocytes; Cattle; Complex; Data; Development; Dissociation; Elements; Engineering; Ensure; Evaluation; Exclusive Breastfeeding; Fermentation; Funding; Glycoproteins; Goals; Grant; Health; Human; Human Development; Human Milk; Immune; Immunity; In Vitro; Infant; Infant Development; Infant Health; Infant formula; Inferior; Intervention; Iron; Knockout Mice; Lactoferrin; Life; Link; Lower Respiratory Tract Infection; Mannose; Marketing; Measures; Mediating; Metabolic; Methods; Milk; Milk Proteins; Modeling; Modernization; Modification; Morbidity - disease rate; Mus; Neurologic; Nutritional; Oral; Parents; Pathway interactions; Peripheral Blood Mononuclear Cell; Permeability; Phenotype; Physiological; Polysaccharides; Procedures; Process; Production; Proliferating; Protein Denaturation; Proteins; Recombinants; Recovery; Regulation; Reporting; Resources; Risk; Role; Safety; Sampling; Signal Transduction; Source; Structure; System; T-Cell Activation; Technology; Testing; United States National Institutes of Health; Work; antimicrobial; commercialization; comparative; cost; cost effective; cytokine; design; efficacy evaluation; environmental stressor; fortification; gastrointestinal infection; glycosylation; gut health; immune function; immunoregulation; improved; in vivo; infant nutrition; intestinal barrier; intestinal maturation; mannose receptor; manufacturing process; microbial; microbial host; microbiome; microbiota; monocyte; mortality; mortality risk; mouse model; nutrition; pre-clinical; prevent; pup; rRNA Genes; response; treatment group; uptake

Project Summary/Abstract Most infants currently rely on infant formula (IF) to meet their nutritional needs. However, existing infant formula is functionally inferior because it lacks breast milk bioactive proteins that are crucial to infant health and development. Consequently, formula-fed infants have increased risk of morbidity and mortality. Current technology limits the ability to add bioactive milk proteins to infant formula because manufacturing processes are resource intensive, costly, and difficult to scale. Thus, scalable technology to improve bioactive availability for fortification of infant formula is needed. Towards this goal, TurtleTree has developed precision fermentation technology to produce recombinant bovine lactoferrin (rbLf), a critical bioactive milk protein demonstrated to support gut health and immune function in infants. Not only will TurtleTree’s technology allow for inclusion of lactoferrin in all infant formulas, but the use of precision fermentation will also dissociate bioactive milk protein production from animal agriculture, as required for cow’s milk-derived lactoferrin (cMDLf), and the associated environmental stressors. In this proposal, we aim to answer two specific questions that will validate our ability to apply precision fermentation technology for infant health and commercialize rbLf as an ingredient for addition to infant formula: 1) Does the unique glycosolation profile of rbLf impact the protein’s immunomodulatory and gut function as compared to cMDLf? 2) Does rbLf improve gut health in an infant preclinical mouse model? Preliminary data supporting this proposal demonstrates the N-linked glycan profile is the only substantially unique structural element of rbLf, characterized by dominance of high-mannose type glycans (~99%) compared to cMDLf (~45%). Thus, we reason a proper evaluation of rbLf requires focused studies on physiological systems sensitive to glycan alterations, notably immune interactions (Aim 1) and gut development (Aim 2). Comparative results between differentially glycosylated lactoferrin treatment groups will distinguish glycan effects. In Aim 1, we will assess the role of lactoferrin N-linked glycosylation on immunomodulation mediated by the mannose receptor in order to understand glycan effect. In Aim 2, we will execute an intervention of orally-delivered rbLf to improve dysregulated gut health in infant mouse pups. With the successful completion of both aims, we expect to demonstrate rbLf promotes immune and gut interactions that maintain biological activity and efficacy as compared to cMDLf. The scope of work is designed to support regulatory approval and expedite TurtleTree’s path to market for the safe and effective application of advancing infant health.

项目摘要/摘要 大多数婴儿目前依靠婴儿配方奶粉(IF)来满足他们的营养需求。然而，现有的婴儿 配方奶粉的功能较差，因为它缺乏对婴儿健康至关重要的母乳生物活性蛋白。 和发展。因此，配方奶婴儿发病和死亡的风险增加。当前 技术限制了在婴儿配方奶粉中添加生物活性乳蛋白的能力，因为制造 流程是资源密集型、成本高昂且难以扩展。因此，可扩展的技术可以提高 婴儿配方奶粉的强化需要生物活性可用性。为了实现这一目标，海龟树已经 开发了一种生产重组牛乳铁蛋白(RbLf)的精密发酵技术 生物活性乳蛋白被证明支持婴儿的肠道健康和免疫功能。不仅会 TurtleTree的技术允许在所有婴儿配方奶粉中包含乳铁蛋白，但使用精密度 发酵还将使生物活性牛奶蛋白的生产从动物农业中分离出来，这是 奶牛乳源乳铁蛋白(CMDLf)和相关的环境应激源。在这项提案中，我们 目的回答两个具体的问题，以验证我们应用精密发酵技术的能力 为了婴儿健康并将rbLf作为添加到婴儿配方奶粉中的成分进行商业化：1)是否具有独特的 RbLf的糖基化对蛋白质的免疫调节和肠道功能的影响 CMDLf？2)rbLf能改善临床前小鼠婴儿模型的肠道健康吗？初步数据支持 这一提议证明了N-连接的糖链轮廓是唯一基本上独特的结构元素 RbLf，以高甘露糖型多糖为主(~99%)，而cMDLf(~45%)。因此， 我们认为，要对rbLf进行适当的评估，需要对对多糖敏感的生理系统进行重点研究。 改变，特别是免疫相互作用(目标1)和肠道发育(目标2)。比较结果 不同的糖基化乳铁蛋白治疗组之间将区分葡聚糖的效果。在目标1中，我们 将评估乳铁蛋白N连接糖基化在甘露糖介导的免疫调节中的作用 为了了解糖链对受体的影响。在目标2中，我们将实施口服分娩的干预 RbLf改善幼鼠肠道功能紊乱的作用。随着这两个目标的成功实现， 我们希望证明rbLf促进免疫和肠道相互作用，从而保持生物活性和 与cMDLf相比的疗效。工作范围旨在支持监管部门的批准和加快 海龟树安全有效地应用于促进婴儿健康的上市之路。