Genetic variation in the metal transporter ZIP8 and impact on the innate immune response in the gut

金属转运蛋白 ZIP8 的遗传变异及其对肠道先天免疫反应的影响

• 批准号: 10310627
• 负责人: Joanna Miller Peloquin Melia
• 金额: $ 5.34万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10310627
• 关键词: Adenoviruses; Advisory Committees; Affect; American; Antigens; Arginine; Attention; Basic Science; Biological Models; Biology; Body Weight; CRISPR/Cas technology; Citrobacter rodentium; Clinical; Colitis; Complement; Complex; Crohn' s disease; DNA Sequence Alteration; Data; Diet; Dietary Intervention; Digestive System Disorders; Disease; Disease Marker; Distal part of ileum; Down-Regulation; Environmental Risk Factor; Epithelial; Epithelial Cells; Equilibrium; Foundations; Functional disorder; Future; Gastroenterology; Gastrointestinal tract structure; Generations; Genes; Genetic; Genetic Variation; Genetic study; Goals; Health; Heterozygote; Homeostasis; Homo; Human; Human Genome; Hypertension; Immune; Immune response; Impairment; In Vitro; Individual; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Innate Immune Response; Institution; Interferon Type II; Intestines; Knock-in Mouse; Literature; Manganese; Mediating; Medicine; Mentorship; Metabolism; Metals; Methods; Micronutrients; Modeling; Molecular Biology; Molecular Immunology; Mus; Mutation; Natural Immunity; Nitric Oxide; Nitric Oxide Synthase; Obesity; Oxidative Stress; Parkinson Disease; Pathogenesis; Pathogenicity; Patients; Phenotype; Physicians; Positioning Attribute; Predisposition; Preparation; Regulation; Research; Role; Schizophrenia; Scientist; Severity of illness; Signal Pathway; Signal Transduction; Single Nucleotide Polymorphism; Stimulus; System; TNF gene; Testing; Tissues; Training; Training and Education; Translational Research; Ulcerative Colitis; Variant; Zinc; Zinc deficiency; antimicrobial peptide; arginase; career; cell type; cytokine; cytotoxicity; didactic education; dietary; disease diagnosis; dysbiosis; enteric infection; experimental study; genome wide association study; host microbiota; immunoregulation; in vivo; in vivo Model; inflammatory disease of the intestine; innate immune mechanisms; innate immune pathways; intestinal epithelium; intestinal homeostasis; knock-down; mRNA Expression; medical schools; microbial; microbiota; mouse model; novel; overexpression; pleiotropism; professor; tool; zinc-binding protein

PROJECT ABSTRACT Inflammatory bowel diseases (IBD), including Crohn’s disease (CD) and ulcerative colitis, affect 1 in 200 Americans. IBD arises through a complex interplay of genetic, immune, microbial, and environmental factors disrupting intestinal homeostasis. There is a strong call from patients to study the role of diet-derived micronutrients in intestinal homeostasis. Recent genetic studies have focused attention on ZIP8 and the role of metal transport in CD through the discovery of an association between CD and a nonsynonymous single nucleotide polymorphism (SNP, rs13107325; Ala391Thr) in SLC39A8, the gene encoding ZIP8. This SNP has also been associated with six other diseases, including obesity and schizophrenia. Further, individuals with ZIP8 A391T shared a common fecal dysbiosis independent of disease diagnosis. The function of ZIP8 in the gut is not known, but in other cell types, ZIP8-mediated zinc transport negatively regulates NF-κB signaling by complexing with IKKβ and ZIP8-mediated manganese transport balances arginine metabolism away from nitric oxide synthase to reduce oxidative stress. Our preliminary data demonstrate that ZIP8 is increased in the inflamed terminal ileum of patients with CD and ZIP8 A391T impairs negative regulation of NF-κB signaling with reduced zinc transport. We have established two model systems for the studies proposed in this application: (i) ZIP8-knockdown in human ileal enteroids and (ii) a novel knock-in mouse with ZIP8 A393T, the mouse equivalent of the human variant. We hypothesize ZIP8-mediated metal transport regulates the innate immune response in intestinal epithelial cells, and this function is changed by ZIP8 A391T to promote CD pathogenesis. The aims of this project are (1) To establish the role of ZIP8 in intestinal epithelial cells in the innate immune response, (2) To study the effect of the CD-associated genetic variation in ZIP8 (A391T) on ZIP8 function in intestinal epithelial cells, and (3) To study the effect of the CD-associated genetic variation in ZIP8 on colitis susceptibility in a novel knock-in mouse model (ZIP8 A393T). The candidate is an Assistant Professor of Medicine at the Johns Hopkins School of Medicine in the Division of Gastroenterology with research and clinical training dedicated to IBD. The goal for this applicant is to use this project to enhance her molecular biology and immunology expertise and position her to build an independent career as a physician- scientist dedicated to studying the role of micronutrients in the pathophysiology of IBD. In addition to hands-on training and didactic education, the Training Plan includes strong mentorship from a Scientific Advisory Committee with diverse expertise in epithelial biology, metal biology, NF-κB signaling, oxidative stress, host- microbiota interactions and IBD, complemented by the strong support of the Hopkins Conte Digestive Diseases Basic and Translational Research Core Center and the institution.

项目摘要 炎症性肠病(IBD)，包括克罗恩病(CD)和溃疡性结肠炎，每200人中就有1人受到影响 美国人。IBD的发生是遗传、免疫、微生物和环境因素复杂相互作用的结果。 扰乱肠道内环境的平衡。患者强烈呼吁研究饮食衍生的作用 肠道内环境平衡中的微量营养素。最近的遗传学研究将注意力集中在ZIP8及其在基因中的作用 通过发现Cd与非同义单链之间的关联来研究Cd中的金属传输 编码ZIP8的基因SLC39A8的SNP，rs13107325；Ala391Thr。这个SNP有 还与其他六种疾病有关，包括肥胖症和精神分裂症。此外，拥有 ZIP8 A391T具有共同的粪便失调，独立于疾病诊断。ZIP8在网络中的作用 肠道是未知的，但在其他类型的细胞中，ZIP8介导的锌转运通过以下方式负向调节NF-κB信号 与IKKβ和ZIP8介导的锰转运络合物平衡精氨酸代谢远离硝酸根 氧化合成酶，以减少氧化应激。我们的初步数据表明，ZIP8在 CD和ZIP8A391T患者回肠末端炎症损害核因子-κB信号的负调节 减少了锌的运输。我们为本文提出的研究建立了两个模型系统 应用：(I)ZIP8-在人回肠肠道中被敲除；(Ii)一种新的带有ZIP8 A393T的敲入小鼠， 老鼠相当于人类的变种。我们假设ZIP8介导的金属转运调节先天的 肠上皮细胞的免疫反应及ZIP8 A391T促进CD的这一功能改变 发病机制。本项目的目的是(1)确定ZIP8在肠上皮细胞中的作用。 先天免疫反应，(2)研究ZIP8(A391T)CD相关基因变异对机体免疫功能的影响 ZIP8在肠上皮细胞中的功能，以及(3)研究Cd相关基因变异对大鼠肠上皮细胞功能的影响。 ZIP8对新型敲入小鼠模型(ZIP8 A393T)结肠炎易感性的影响。应聘者是助理 约翰霍普金斯大学医学院消化系医学教授 致力于IBD的研究和临床培训。这位申请者的目标是利用这个项目提升她的能力 分子生物学和免疫学专业知识，并使她成为一名独立的内科医生- 致力于研究微量营养素在IBD病理生理学中的作用的科学家。除了亲身实践之外 培训和教育，培训计划包括来自科学咨询的强有力的指导 委员会在上皮生物学、金属生物学、核因子-κB信号转导、氧化应激、宿主- 微生物区系相互作用和IBD，并得到霍普金斯大学消化疾病中心的大力支持 基础和翻译研究核心中心和机构。