Fusobacterial Associated Beta Defensin Inducer

梭杆菌相关β防御素诱导剂

• 批准号: 9326495
• 负责人: AARON WEINBERG
• 金额: $ 47.51万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9326495
• 关键词: Address; Alanine; Antibiotics; Bacteria; CCL20 gene; Categories; Cell Wall; Cell surface; Cells; Characteristics; Chronic; Complex; Cysteine; Data; Development; Diglycerides; Disease; Epithelial Cells; Figs - dietary; Fusobacterium nucleatum; Future; Gene Expression; Genes; Gram-Negative Bacteria; Healed; Health; Heterodimerization; Human; Immune response; In Situ; Infection; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Inflammatory Response; Intention; Intervention; Invaded; Knowledge; Lactobacillus; Lipoproteins; MAPK8 gene; Manuscripts; Medical; Microbe; Mucous Membrane; Mus; NF-kappa B; Ontology; Oral; Oral cavity; Oral mucous membrane structure; Orthologous Gene; Outcome; Peptides; Porphyromonas gingivalis; Positioning Attribute; Probiotics; Procedures; Progress Reports; Property; Protease Inhibitor; Proteins; Reporting; Research; Response Elements; Site; Skin; Surface; System; TLR1 gene; TLR2 gene; Testing; Therapeutic Agents; Tissues; Toxic effect; Transcription Factor AP-1; Ubiquitin; Up-Regulation; Vagina; Wound Healing; antimicrobial; antimicrobial peptide; beta-Defensins; cathelicidin; chemokine; corneal epithelium; cytokine; defense response; gastrointestinal; healing; improved; in vitro testing; in vivo; keratinocyte; microbial; mouse model; mucosal site; multicatalytic endopeptidase complex; novel; novel therapeutics; pre-clinical; secondary infection; translational study

 DESCRIPTION (provided by applicant): Human beta defensins (hBDs) are first-line defenders of the host's mucosa against microbial challenges. We were first to report them in Human Oral Epithelial Cells (HOECs) and demonstrated their antimicrobial and immunoregulatory properties as agents that alter inflammatory responses. We believe that harnessing these and other antimicrobial peptides (AMPs) correctly, i.e., promoting their expression in vulnerable sites, could impact the course of infection and wound repair. We have isolated and characterized a cell wall associated molecule from F. nucleatum ATCC 25586 that is principally responsible for hBD induction and refer to it as FAD-I for F. nucleatum Associated beta Defensin Inducer. We have identified that the molecule's activity resides in the cysteine-diacylglycerol complex at position 16, as alanine substitution abrogates activity, and that hBD2 induction is dependent upon interaction of FAD-I with TLRs 1 and 2 on the HOEC surface. Other AMPs are also induced by FAD-I, including Mip3 (CCL20) and LL37 which are elevated in epithelial cells following FAD-I challenge. Moreover, our preliminary data suggests that FAD-I's induction of hBD-2 happens with little simultaneous induction of pro- inflammatory cytokines. Functionally, HOECs challenged with FAD-I are protected from invasion by Porphyromonas gingivalis, a major etiologic agent in the initiation of periodontal destruction. Therefore, our overarching hypothesis is that commensal bacterial molecules, such as FAD-I, that promote the expression of beneficial innate response elements without inflammation, could be harnessed to protect tissues from microbial challenges and inflammatory damage. In order to develop FAD-I into a novel therapeutic agent, we will address several existing knowledge gaps including in vivo tests of FAD-I in various mouse mucosal tissues to determine its' ability to induce in situ AMP expression as well as develop a novel vehicle for delivery of FAD-I for intervention in disease modulation. Our aims are intended to: (1) Determine key characteristics of FAD-I that result in stimulation of epithelial cell-derived AMPs in the absence of an inflammatory response; (2) Determine FAD-I's ability to induce mouse AMP orthologs and quantify pre-clinical FAD-I potency in vivo using the pre-clinical murine models of wound healing and inflammation; (3) Construct a FAD-I producing Lactobacillus for probiotic in vitro testing. Our overall intention is to prepare ourselves to use FAD-I for future translational studies in modulating dysbiosis and chronic inflammation in defined diseased mouse models, which may pave the way for interventional studies in humans.

 描述（由申请人提供）：人类β防御素（hBD）是宿主粘膜抵御微生物挑战的一线防御者，我们首先在人类口腔上皮细胞（HOEC）中报告了它们，并证明了它们作为药物的抗菌和免疫调节特性。我们相信，正确利用这些和其他抗菌肽（AMP），即促进它们在脆弱部位的表达，可以改变炎症反应。我们从具核梭菌 ATCC 25586 中分离并表征了主要负责 hBD 诱导的细胞壁相关分子，并将其称为具核梭菌相关 β 防御素诱导剂的 FAD-I。已经确定该分子的活性存在于位置 16 的半胱氨酸-二酰基甘油复合物中，因为丙氨酸取代会消除活性，并且 hBD2 诱导依赖于 FAD-I 与 HOEC 表面上的 TLR 1 和 2 的相互作用。FAD-I 也会诱导其他 AMP，包括在 FAD-I 攻击后在上皮细胞中升高的 Mip3 (CCL20) 和 LL37。我们的初步数据表明，FAD-I 对 hBD-2 的诱导很少同时诱导促炎性细胞因子。牙龈卟啉单胞菌是引发牙周破坏的主要病原体，因此，我们的总体假设是，可以利用共生细菌分子，例如 FAD-I，促进有益先天反应元件的表达，而不会产生炎症。保护组织免受微生物挑战和炎症损伤 为了将 FAD-I 开发成一种新型治疗剂，我们将解决几个现有的知识空白，包括在各种小鼠中进行 FAD-I 的体内测试。粘膜组织，以确定其诱导原位 AMP 表达的能力，并开发一种用于递送 FAD-I 以干预疾病调节的新型载体。我们的目标是：(1) 确定所产生的 FAD-I 的关键特征。在没有炎症反应的情况下刺激上皮细胞衍生的 AMP；(2) 确定 FAD-I 诱导小鼠 AMP 直向同源物的能力并量化临床前 FAD-I 体内效力；使用伤口愈合和炎症的临床前小鼠模型；（3）构建用于益生菌体外测试的FAD-I乳酸菌。 准备使用 FAD-I 进行未来的转化研究，在特定的患病小鼠模型中调节生态失调和慢性炎症，这可能为人类的介入研究铺平道路。