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: 10544183
• 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/10544183
• 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; Cells; Characteristics; Chemicals; Chronic; Clinical; Colitis; Colon; Communities; Crohn' s disease; Data; Development; Disease; Disease model; Distal; Doctor of Philosophy; Enteric Nervous System; Enterobacteria phage P1 Cre recombinase; Enteroendocrine Cell; Enzymes; Epithelium; Excision; FOLH1 gene; Foundations; Ganglia; Gastrointestinal Diseases; Gastrointestinal tract structure; Generations; Genetic; 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; Sulfonic Acids; TNF gene; Tamoxifen; Technical Expertise; Testing; Therapeutic; Thick; Time; Training; Transgenic Organisms; Treatment Failure; Ulcerative Colitis; United States; Up-Regulation; Veterinarians; Visceral pain; Work; Zinc; antagonist; career; cell type; 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; nitrobenzene; novel; overexpression; pre-clinical; preclinical study; programs; promoter; protein expression; response; selective expression; small molecule; 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.

项目概要/摘要 K01 SERCA 奖项将为 Diane Peters（DVM、MS、PhD）提供受保护的研究时间和指导 她建立了独立的生物医学研究生涯。接受过兽医双重培训 药理学家 Peters 博士在比较医学、人类动物模型方面拥有深厚的背景 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 根据该提案的概述，她将扩展自己对炎症性肠病 (IBD) 的了解，并建立一个 先进的技术技能，包括免疫荧光成像、CRISPR/Cas9 基因组编辑和 胃肠道表型分析方法，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，并与高死亡率相关。 公共卫生负担。 IBD 无法治愈，并且很大一部分受影响的个体没有反应 所有可用的治疗方法。谷氨酸羧肽酶 II (GCPII) 是一种高度高效的锌金属肽酶 在 IBD 的两种主要亚型中过度表达：克罗恩病和溃疡性结肠炎。 GCPII 的承诺 作为 IBD 的治疗靶点已在多项独立的临床前研究中得到证实，这些研究 研究表明，小分子 GCPII 抑制剂在三种机制不同的药物中具有显着的抗结肠炎活性 鼠标模型。虽然很明显 GCPII 上调与人类和小鼠 IBD 相关， 关于其在疾病中的功能存在知识差距。 K01 SERCA 研究将产生重要的新数据 与 IBD 中 GCPII 的生物学相关。具体来说，在目标 #1 中，GCPII 表达式将在正常和 IBD 影响人类和小鼠 IBD 的胃肠组织。在目标#2中，一种新颖的敲入小鼠将是 生成在回肠和结肠中过度表达人 GCPII，然后进行纵向表征 屏障功能、胃肠道炎症、微生物组成和内脏疼痛反应。这是 假设 GCPII 过度表达的定位在人和小鼠 IBD 中是保守的，并且 强制回肠和结肠 GCPII 过度表达的成年小鼠会自发发展结肠炎。 成功完成这些研究目标将 (1) 增加我们对 GCPII 失调的理解 IBD，(2) 识别与正在进行的药物开发工作相关的 GCPII+ 靶细胞群，以及 (3) 产生了一种新的 IBD 小鼠模型，该模型可能与人类疾病具有更高的相似性。拟议的 研究将由专家科学家和临床医生组成的多学科团队监督，其中包括： Barbara Slusher，约翰·霍普金斯大学药物发现部主任，Pankaj Jay Pasricha 博士，约翰·霍普金斯大学主任 霍普金斯大学神经胃肠病学中心，辛西娅·西尔斯博士，约翰·霍普金斯大学无菌中心主任 Murine Core，Robert Anders 博士，胃肠道病理学家，Christine McDonald 博士，专家 胃肠道屏障功能，以及胃肠道病理学家和专家 Thaddeus Stappenbeck 博士 Mucosal Immunity，他们非常适合指导 Diane Peters 博士的 IBD 转化研究生涯。