Hyaluronan regulation of microbial host defense of the intestine

透明质酸对肠道微生物宿主防御的调节

• 批准号: 8511751
• 负责人: Carol A. de la Motte
• 金额: $ 38.54万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-15 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8511751
• 关键词: Adhesions; Age; Animals; Antibodies; Antiviral Agents; Bacteria; Bacteriophages; Base Composition; Bifidobacterium; Binding; Biological; Breast Feeding; CD44 Antigens; Carbohydrates; Cattle; Cell Culture Techniques; Cell Line; Child; Colitis; Communicable Diseases; Complex; Data; Defense Mechanisms; Defensins; Dendritic Cells; Development; Diet; Disaccharides; Environment; Epithelial; Epithelial Cells; Epithelium; Family; Food Supplements; Gastrointestinal tract structure; Glucosamine; Glucuronic Acids; Growth; Health; Health Food; Host Defense; Human; Human Milk; Hyaluronan; Immune; Immune response; In Vitro; Individual; Infant; Infant formula; Infection; Inflammatory disease of the intestine; Intestines; Investigation; Lactation; Lead; Ligands; Measures; Mediating; Medical; Milk; Modeling; Molecular Weight; Mus; Necrotizing Enterocolitis; Newborn Infant; Oligosaccharides; Operative Surgical Procedures; Pathologic; Pathway interactions; Polymers; Polysaccharides; Prevention; Production; Property; Proteins; Regulation; Replacement Arthroplasty; Reporting; Skin; Specificity; Staging; Structure; System; TLR2 gene; TLR4 gene; Testing; Therapeutic; Time; Toll-like receptors; Up-Regulation; Viral; Viral Genes; Virus; Virus Diseases; antimicrobial; antimicrobial peptide; base; carbohydrate structure; design; feeding; gastrointestinal; immunogenic; in vivo; intestinal epithelium; member; microbial host; mouse model; pathogen; prevent; prophylactic; protective effect; protein expression; receptor; response; sensor; sugar; toll-like receptor 4; treatment strategy

DESCRIPTION (provided by applicant): Breastfeeding of human infants has been shown to provide protection against many pathogenic and pathologic conditions, and much of that protection is mediated by naturally occurring glycans in milk. We have recently identified a new glycan, hyaluronan (HA), in human milk, which occurs at the highest concentration during the early stage of human lactation and decreases gradually over time. Human milk HA levels are markedly higher than in commercial infant formulas. Importantly, we have also discovered that feeding purified HA of a specific size range to mice is protective in an epithelial damage/ bacteria-induced colitis model. Additionally our data indicate size-specific, HA fragment driven anti-microbial responses by intestinal epithelium. A special attribute of HA, different from other milk glycans, is its function as an endogenous ligand of the bacterial recognition Toll-like receptors (TLRs) 4 and 2, which are known to be important sensors that trigger the innate host defense system. Thus HA may be a way to stimulate the uncolonized infant gut to protect against pathogens. Therefore we hypothesize that HA in milk, in a size dependent manner, provides innate host protection against pathogens in the gut of immune-naive infants and children. Preliminary studies from our lab show HA size-specific upregulation of antimicrobial responses in intestinal epithelial cells in cell cultures and mouse models. Among the virus response proteins, our data show IFIT1 is markedly induced by HA, specifically of the 35kDa size range, and a combination of antibodies to TLR4 and TLR2 completely blocks this effect. Importantly, the HA we can isolate from milk also has the same stimulatory ability. The HA 35kDa fragments also specifically induce human 2-defensin 2, one member of the antimicrobial peptide family that is important in excluding pathogens and maintaining proper intestinal bacterial colonization. Again, the HA isolated from human milk has the same ability. We envision that our proposed investigation could ultimately lead to identifying defined dietary HA supplements that would be therapeutic to infants with NEC, infants who are formula fed and children prone to gastrointestinal infectious disease. HA would be an attractive therapeutic because it is: 1) not toxic or immunogenic due to its simple unmodified carbohydrate structure; 2) easily administered; 3) already being produced commercially on a large scale as a health food supplement as well as for medical applications. Based on our preliminary data and the previously reported biological effects of HA, we think the key to its protective potency will depend on the HA polysaccharide size(s) needed to mediate individual host responses.

描述（由申请人提供）：母乳喂养人类婴儿已被证明可以提供针对许多致病和病理状况的保护，并且大部分保护是由牛奶中天然存在的聚糖介导的。我们最近在母乳中发现了一种新的聚糖，即透明质酸（HA），它在人类哺乳的早期阶段浓度最高，并随着时间的推移逐渐降低。母乳中的 HA 水平明显高于商业婴儿配方奶粉。重要的是，我们还发现，给小鼠喂食特定大小范围的纯化 HA 对上皮损伤/细菌诱导的结肠炎模型具有保护作用。此外，我们的数据表明大小特异性、HA 片段驱动肠上皮的抗微生物反应。 HA 与其他乳聚糖不同的一个特殊属性是其作为细菌识别 Toll 样受体 (TLR) 4 和 2 的内源配体的功能，众所周知，TLR 是触发先天宿主防御系统的重要传感器。因此，HA 可能是刺激未定植婴儿肠道以抵御病原体的一种方法。因此，我们假设牛奶中的透明质酸以大小依赖性方式为未免疫婴儿和儿童的肠道中的病原体提供先天宿主保护。我们实验室的初步研究表明，在细胞培养物和小鼠模型中，HA 大小特异性上调肠上皮细胞的抗菌反应。在病毒反应蛋白中，我们的数据显示 IFIT1 明显被 HA 诱导，特别是 35kDa 大小范围的 HA，而 TLR4 和 TLR2 抗体的组合完全阻断了这种效应。重要的是，我们从牛奶中分离出来的透明质酸也具有相同的刺激能力。 HA 35kDa 片段还特异性诱导人 2-防御素 2，它是抗菌肽家族的成员之一，对于排除病原体和维持适当的肠道细菌定植非常重要。同样，从母乳中分离出的透明质酸也具有相同的能力。我们预计，我们提出的研究最终可能会确定特定的膳食透明质酸补充剂，这些补充剂对患有 NEC 的婴儿、配方奶喂养的婴儿和易患胃肠道传染病的儿童具有治疗作用。 HA 将是一种有吸引力的治疗方法，因为它： 1) 由于其简单的未修饰的碳水化合物结构，无毒性或免疫原性； 2）易于管理； 3) 已经作为保健食品补充剂以及医疗应用进行大规模商业化生产。根据我们的初步数据和之前报道的 HA 生物效应，我们认为其保护效力的关键将取决于介导个体宿主反应所需的 HA 多糖大小。

项目成果

Hyaluronan Synthase 3 Null Mice Exhibit Decreased Intestinal Inflammation and Tissue Damage in the DSS-Induced Colitis Model.

• DOI: 10.1155/2015/745237
• 发表时间: 2015
• 期刊: International journal of cell biology
• 作者: Kessler SP;Obery DR;de la Motte C
• 通讯作者: de la Motte C

Agarose and polyacrylamide gel electrophoresis methods for molecular mass analysis of 5- to 500-kDa hyaluronan.

• DOI: 10.1016/j.ab.2011.05.026
• 发表时间: 2011-10-01
• 期刊: Analytical biochemistry
• 影响因子: 2.9
• 作者: Bhilocha S;Amin R;Pandya M;Yuan H;Tank M;LoBello J;Shytuhina A;Wang W;Wisniewski HG;de la Motte C;Cowman MK
• 通讯作者: Cowman MK

The role of hyaluronan in innate defense responses of the intestine.

• DOI: 10.1155/2015/481301
• 发表时间: 2015
• 期刊: International journal of cell biology
• 作者: de la Motte CA;Kessler SP
• 通讯作者: Kessler SP

Determination of hyaluronan molecular mass distribution in human breast milk.

• DOI: 10.1016/j.ab.2014.12.020
• 发表时间: 2015-04-01
• 期刊: ANALYTICAL BIOCHEMISTRY
• 影响因子: 2.9
• 作者: Yuan, Han;Amin, Ripal;Ye, Xin;de la Motte, Carol A.;Cowman, Mary K.
• 通讯作者: Cowman, Mary K.