Generation and characterization of a humanized mouse model of Crohn's disease

克罗恩病人源化小鼠模型的生成和表征

• 批准号: 10195523
• 负责人: Richard A. Flavell
• 金额: $ 8.38万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10195523
• 关键词: Affect; American; Anemia; Animal Model; Antigen-Antibody Complex; B-Lymphocytes; Blood; CD34 gene; CD4 Positive T Lymphocytes; CRISPR/Cas technology; Cells; Chemicals; Chronic; Clinic; Clinical Trials; Colitis; Colon; Communities; Complex; Crohn' s disease; Development; Disease; Distal part of ileum; Engineering; Engraftment; Environmental Risk Factor; Flow Cytometry; Future; Generations; Genes; Genetic; Genetic Engineering; Goals; Hematopoietic stem cells; Histopathology; Human; Human Genetics; Immune; Immune response; Immune system; Immunodeficient Mouse; Infiltration; Inflammation Mediators; Inflammatory; Inflammatory Bowel Diseases; Intestines; Kinetics; Knock-in; Lamina Propria; Modeling; Molecular; Mus; Myelogenous; Natural History; Natural Killer Cells; Pathogenesis; Pathologic; Pathology; Pathway interactions; Patients; Phenotype; Physiology; Pilot Projects; Population; Preclinical Testing; Regimen; Research; Role; Small Intestines; System; T-Lymphocyte; Technology; Time; Tissues; Transcript; Translating; Ulcerative Colitis; Ursidae Family; confocal imaging; cytokine; effectiveness evaluation; genetic variant; gut microbiota; human imaging; humanized mouse; immunopathology; in vivo; insight; intraepithelial; macrophage; mouse development; mouse model; neutrophil; novel therapeutics; overexpression; pre-clinical; pre-clinical assessment; secondary lymphoid organ; side effect; therapeutic candidate; therapeutic target; tool; translational study

Generation and characterization of a humanized mouse model of Crohn’s disease Crohn’s-disease (CD) and ulcerative colitis (UC) are the two major forms of inflammatory bowel disease (IBD), a chronic inflammatory disorder that affects an estimated 3 million Americans. The etiopathogenesis of CD is distinctive for each patient and involves a complex interaction between genetic factors, the immune system, the gut microbiota and other environmental factors. Mouse models of IBD have been invaluable in providing critical mechanistic insights into its complex etiopathogenesis and in indicating new potential therapeutic targets. However, it has been particularly difficult to translate advances in our understanding of IBD pathogenesis into novel therapies that prove successful in the clinic. A major obstacle is that mice and humans, despite their similarities, have fundamental differences in their intestinal physiology and, crucially, in the function of their immune system. A second problem is that the vast majority of mouse models of IBD recapitulate aspects of the natural history of UC but not the unique pathology of CD. Not surprisingly, notable examples of clinical trials that failed in CD patients were originally guided by mouse models of colitis at the preclinical level. One inviting avenue for pre-clinical testing is humanized mouse modeling, where human hematopoietic stem cells (HSCs) are engrafted into immunodeficient mice and a human immune system develops in the mouse host. Humanized mouse models allow to study the function of human immune cells in vivo without directly involving human patients. Despite recent progress in the humanization of the immune system in mice, the development of humanized models of IBD is lagging behind and has been limited to the chemical induction of colitis upon transfer of human CD4+ T-cells. Translational animal models that faithfully recapitulate human CD pathology have been lacking. In this project we aim to develop the first humanized mouse model of CD. For this purpose we will integrate two state-of-the-art mouse models: a genetic mouse model that specifically resembles the pathological and mechanistic features of human CD and the MISTRG6 humanized model which supports the most comprehensive humanization of the immune system achieved to date, including not only T, B and NK cells, but also myeloid populations crucial in CD such as macrophages and neutrophils. Major goals of this pilot project are to 1) generate these mice by CRISPR/Cas9 engineering and optimize the humanization of their immune system and 2) determine the kinetics of humanized Crohn’s pathogenesis and thoroughly characterize its phenotypic aspects at the level of tissue pathology and intestinal immune responses. This humanized system will be an important and unique preclinical/translational tool for the CD research community. It will reveal pathways employed by human immune cells to drive CD pathogenesis and also enable future studies assessing the effectiveness of candidate therapeutics against human targets as well as studies aiming to uncover the specific effect of human genetic variants associated with CD on the function of a human immune system.

克罗恩病人源化小鼠模型的生成和表征 克罗恩病 (CD) 和溃疡性结肠炎 (UC) 是炎症性肠病的两种主要形式 (IBD)，一种影响大约 300 万美国人的慢性炎症性疾病。发病机制 每个患者的 CD 的发病情况都是不同的，并且涉及遗传因素、免疫因素之间复杂的相互作用。 系统、肠道微生物群和其他环境因素。 IBD 小鼠模型在以下方面具有无价的价值： 为其复杂的发病机制提供关键的机制见解并表明新的潜力 治疗目标。然而，将我们理解的进展转化为特别困难的 IBD 发病机制转化为在临床上证明成功的新疗法。一个主要障碍是老鼠和 尽管人类有相似之处，但其肠道生理学却存在根本差异，最重要的是， 他们的免疫系统的功能。第二个问题是绝大多数 IBD 小鼠模型 概括了 UC 自然史的各个方面，但没有概括 CD 的独特病理学。毫不奇怪， 在 CD 患者中失败的临床试验的著名例子最初是由小鼠结肠炎模型指导的 在临床前水平。临床前测试的一种诱人途径是人性化小鼠建模，其中 将人类造血干细胞（HSC）移植到免疫缺陷小鼠体内，并产生人类免疫细胞 系统在鼠标宿主中开发。人源化小鼠模型可以研究人类的功能 体内免疫细胞，无需直接涉及人类患者。尽管最近在人性化方面取得了进展 在小鼠免疫系统方面，IBD人源化模型的开发相对滞后，并且 仅限于人类 CD4+ T 细胞转移后化学诱导结肠炎。翻译动物 一直缺乏忠实再现人类 CD 病理学的模型。在这个项目中，我们的目标是开发 第一个 CD 人源化小鼠模型。为此，我们将集成两个最先进的鼠标 模型：一种遗传小鼠模型，特别类似于以下疾病的病理和机制特征： human CD 和 MSTRG6 人源化模型，支持最全面的人源化 迄今为止实现的免疫系统，不仅包括 T、B 和 NK 细胞，还包括骨髓细胞群 巨噬细胞和中性粒细胞等在 CD 中至关重要。该试点项目的主要目标是 1) 这些小鼠通过 CRISPR/Cas9 工程改造并优化其免疫系统的人性化以及 2) 确定人源化克罗恩病发病机制的动力学并彻底表征其表型 组织病理学和肠道免疫反应水平方面。这个人性化的系统将是 CD 研究界重要且独特的临床前/转化工具。它将揭示路径 人类免疫细胞利用它来驱动 CD 发病机制，也使未来的研究能够评估 针对人类靶标的候选疗法的有效性以及旨在揭示的研究 与 CD 相关的人类遗传变异对人类免疫系统功能的特定影响。