Exploring Glutamate Carboxypeptidase II (GCPII) Dysregulation in Human and Experimental IBD

探索人类和实验 IBD 中的谷氨酸羧肽酶 II (GCPII) 失调

• 批准号: 10370517
• 负责人: Diane E Peters
• 金额: $ 13.01万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10370517
• 关键词: Acute; Adoptive Transfer; Adult; Affect; Age; Animal Model; Applications Grants; Award; Bacteroides fragilis; Biological Products; Biology; Biomedical Research; Biopsy; CRISPR/Cas technology; Cecum; Cells; Characteristics; Chronic; Clinical; Colitis; Colon; Communities; Crohn' s disease; Data; Development; Dextrans; Disease; Disease model; Distal; Doctor of Philosophy; Enteric Nervous System; Enterobacteria phage P1 Cre recombinase; Enteroendocrine Cell; Enzymes; Epithelial; Excision; FOLH1 gene; Foundations; Ganglia; Gastrointestinal Diseases; Gastrointestinal tract structure; Generations; Genomics; Germ-Free; Homeobox; Human; Ileitis; Image; Immunofluorescence Immunologic; Immunohistochemistry; Individual; Inflammation; Inflammatory Bowel Diseases; Inflammatory Infiltrate; Interleukin-10; Knock-in Mouse; Knockout Mice; Knowledge; Lesion; LoxP-flanked allele; Medicine; Mentors; Metalloproteases; Methodology; Methods; Modeling; Mucosal Immunity; Mucous Membrane; Mus; Operative Surgical Procedures; Pathogenesis; Pathologist; Patients; Pattern; Peptide Hydrolases; Persons; Pharmacology; Phenotype; Pilot Projects; Population; Predisposition; Public Health; Quality of life; Refractory; Reporting; Research; Research Personnel; Resistance; Role; Sampling; Scientist; Solid; Spontaneous colitis; Stains; Sulfonic Acids; Supervision; TNF gene; Tamoxifen; Technical Expertise; Testing; Therapeutic; Thick; Time; Training; Transgenic Organisms; Treatment Failure; Trinitrobenzenes; Ulcerative Colitis; United States; Up-Regulation; Veterinarians; Visceral pain; Work; Zinc; antagonist; career; cell type; chemical genetics; comparative; dextran sulfate sodium induced colitis; disorder subtype; drug development; drug discovery; gastrointestinal; gastrointestinal epithelium; genome editing; human disease; human model; human tissue; ileum; improved; in vivo; inhibitor; insight; microbiome composition; mouse model; multidisciplinary; novel; overexpression; pre-clinical; preclinical study; programs; promoter; protein expression; response; selective expression; small molecule; sodium sulfate; success; therapeutic target

PROJECT SUMMARY/ABSTRACT This K01 SERCA award will provide protected research time and mentoring to Diane Peters, DVM, MS, PhD as she establishes an independent biomedical research career. Dual-trained as a veterinarian and pharmacologist, Dr. Peters has a strong background in comparative medicine, animal models of human disease, in vivo pharmacology, and protease biology. Through completion of the training and research aims outlined in this proposal, she will expand her knowledge of inflammatory bowel disease (IBD) and build an advanced technical skill set, to include immunofluorescent imaging, CRISPR/Cas9 genome editing and gastrointestinal phenotyping methods, that will form a solid foundation for her planned independent research program. IBD is a chronic condition that negatively impacts patient quality of life and is associated with a high public health burden. There is no cure for IBD and a large percentage of affected individuals are unresponsive to all available treatments. Glutamate carboxypeptidase II (GCPII) is zinc metallopeptidase that is highly overexpressed in the two main subtypes of IBD: Crohn's disease and ulcerative colitis. The promise of GCPII as a therapeutic target in IBD has been demonstrated in multiple independent preclinical studies, which have shown that small molecule GCPII inhibitors have significant anti-colitis activity in three mechanistically-distinct mouse models. While it is apparent that GCPII upregulation is relevant in both human and mouse IBD, knowledge gaps exist regarding its function in disease. This K01 SERCA research will yield critical new data relevant to the biology of GCPII in IBD. Specifically, in Aim #1 GCPII expression will be defined in normal and IBD-affected gastrointestinal tissues of human and mouse IBD. In Aim #2, a novel knock-in mouse will be generated that overexpresses human GCPII in the ileum and colon, followed by longitudinal characterization of barrier function, gastrointestinal inflammation, microbiome composition and visceral pain response. It is hypothesized that the localization of GCPII overexpression will be conserved in human and mouse IBD and that adult mice with forced ileum and colon GCPII overexpression will spontaneously develop colitis. Successful completion of these research aims will (1) increase our understanding of GCPII dysregulation in IBD, (2) identify GCPII+ target cell populations with relevance to ongoing to drug development efforts and (3) yield a novel mouse model of IBD that may have increased similarity to human disease. The proposed research will be supervised by a multidisciplinary team of expert scientists and clinicians including: Dr. Barbara Slusher, Director of Johns Hopkins Drug Discovery, Dr. Pankaj Jay Pasricha, Director of the Johns Hopkins Center for Neurogastroenterology, Dr. Cynthia Sears, Director of the Johns Hopkins Germ Free Murine Core, Dr. Robert Anders, Gastrointestinal Pathologist, Dr. Christine McDonald, expert in Gastrointestinal Barrier Function, and Dr. Thaddeus Stappenbeck, Gastrointestinal Pathologist and expert in Mucosal Immunity, who are well-suited to mentor Dr. Diane Peters in her translational IBD research career.

项目摘要/摘要 这项K01 SERCA奖将为Diane Peters，DVM，MS，PhD提供受保护的研究时间和指导 因为她建立了独立的生物医学研究事业。接受过兽医和兽医双重培训 药理学家，彼得斯博士在比较医学，人类动物模型方面有很强的背景 疾病、体内药理学和蛋白酶生物学。通过完成培训和研究目标 在这项提案中，她将扩大她对炎症性肠病(IBD)的知识，并建立一个 高级技术技能，包括免疫荧光成像、CRISPR/Cas9基因组编辑和 胃肠道表型方法，这将为她计划的独立研究奠定坚实的基础 程序。IBD是一种慢性疾病，会对患者的生活质量产生负面影响，并与高 公共卫生负担。目前还没有治愈IBD的方法，而且很大一部分受感染的人没有反应。 所有可用的治疗方法。谷氨酸羧肽酶II(GCPII)是一种高活性的锌金属肽酶。 在IBD的两个主要亚型：克罗恩病和溃疡性结肠炎中过度表达。GCPII的承诺 作为IBD的治疗靶点，已经在多项独立的临床前研究中得到证明，这些研究 显示小分子GCPII抑制剂在三种不同的机制中具有显著的抗结肠炎活性 老鼠模型。虽然很明显GCPII上调在人和小鼠IBD中都相关， 关于它在疾病中的作用，存在着知识空白。这项K01 SERCA研究将产生关键的新数据 与IBD的GCPII生物学相关。具体地说，在AIM#1中，GCPII的表达将以正常和 IBD感染的人和小鼠的胃肠道组织。在Aim#2中，一种新的敲入鼠标将是 在回肠和结肠中过度表达人GCPII，然后纵向表征 屏障功能、胃肠道炎症、微生物群组成和内脏疼痛反应。它是 假设GCPII过度表达的定位在人和小鼠IBD和 强迫回肠和结肠GCPII过度表达的成年小鼠将自发发展为结肠炎。 这些研究目标的成功完成将(1)增加我们对GCPII失调的理解 IBD，(2)确定与正在进行的药物开发努力相关的GCPII+目标细胞群，以及(3) 产生一种新的IBD小鼠模型，该模型可能增加了与人类疾病的相似性。建议数 研究将由一个由专家科学家和临床医生组成的多学科团队进行监督，其中包括： Barbara Slusher，Johns Hopkins药物发现主任，Pankaj Jay Pasricha博士，Johns主任 霍普金斯神经胃肠病中心，辛西娅·西尔斯博士，约翰·霍普金斯无菌中心主任 小鼠核心，罗伯特·安德斯博士，胃肠病理学家，克里斯汀·麦克唐纳博士， 胃肠道病理学家和胃肠疾病专家塞迪厄斯·斯塔彭贝克博士 粘膜免疫，他们非常适合在黛安·彼得斯博士的翻译性IBD研究生涯中指导她。