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 该K01 SERCA奖将为Diane Peters，DVM，MS，PhD提供受保护的研究时间和指导 当她建立独立的生物医学研究生涯时。双重训练作为兽医和 药物学家彼得斯博士在比较医学上具有强大的背景，人类动物模型 疾病，体内药理学和蛋白酶生物学。通过完成培训和研究目的 在此提案中概述了她将扩大对炎症性肠病（IBD）的了解，并建立 先进的技术技能集，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构成坚实的基础 程序。 IBD是一种慢性病，会对患者的生活质量产生负面影响，并且与高有关 公共卫生负担。 IBD无法治愈，很大一部分受影响的人没有反应 对所有可用的治疗。谷氨酸羧肽酶II（GCPII）是高度高的锌金属肽酶 在IBD的两个主要亚型中过表达：克罗恩病和溃疡性结肠炎。 GCPII的承诺 作为IBD的治疗靶标在多个独立的临床前研究中已证明 表明小分子GCPII抑制剂在三个机械上的抗癌性活性显着 鼠标模型。虽然显然是GCPII上调在人和小鼠IBD中都相关，但 知识差距就其在疾病中的功能存在。这项K01 SERCA研究将产生关键的新数据 与IBD中GCPII的生物学有关。具体而言，在AIM＃1中，GCPII表达式将在正常和 人和小鼠IBD的IBD影响的胃肠道组织。在AIM＃2中，一种新颖的敲击鼠标将是 产生了过表达回肠和结肠中的人类GCPII，然后进行纵向表征 屏障功能，胃肠道炎症，微生物组组成和内脏疼痛反应。这是 假设GCPII过表达的定位将在人类和小鼠IBD中保存下来，以及 那些具有强迫回肠和结肠GCPII过表达的成年小鼠会自发发展结肠炎。 这些研究目标的成功完成将（1）提高我们对GCPII失调的理解 IBD，（2）识别GCPII+目标细胞群，与持续的药物开发工作有关，（3） 产生一种新型的IBD小鼠模型，可能与人类疾病相似。提议 研究将由专家科学家和临床医生组成的跨学科团队进行监督： 约翰·霍普金斯（Johns Hopkins）毒品发现的董事芭芭拉·斯拉瑟（Barbara Slusher） 霍普金斯神经胃肠病学中心，约翰·霍普金斯·杰姆的主任辛西娅·西尔斯博士 Murine Core，Robert Anders博士，胃肠道病理学家，Christine McDonald博士，专家 胃肠道障碍功能和胃肠道病理学家Thaddeus Stappenbeck博士兼专家 粘膜免疫力，非常适合指导戴安娜·彼得斯（Diane Peters）博士的翻译IBD研究生涯。