Enhancement of Human Immune System Development in Mouse Models

增强小鼠模型中的人类免疫系统发育

基本信息

批准号：
10652645
负责人：
Santhi Gorantla
金额：
$ 19.19万
依托单位：
UNIVERSITY OF NEBRASKA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10652645
关键词：
Address Animal Model Animals B-Lymphocytes Basic Science Binding Proteins Biological Process Biomedical Research Birth CD34 gene CRISPR/Cas technology CXCL13 gene Cell Communication Cell Separation Cells Clinical Trials Communities Conceptions Cytokine Signaling Dedications Development Disease Distant Engineering Engraftment Enzyme Induction Experimental Models Family Fumarylacetoacetase Genetic Genetic Transcription Genetically Engineered Mouse Genetically Modified Animals Genome engineering Goals Growth HIV HIV-1 Health Hematopoietic Hematopoietic Stem Cell Transplantation Hematopoietic stem cells Hepatitis B Hepatitis B Infection Hepatitis B Vaccination Hepatitis C virus Hepatitis Viruses Hepatocyte Homologous Gene Human Hydrolase IL7 gene Immune Immune response Immune system Immunity Inbred BALB C Mice Infection Interleukin 2 Receptor Gamma Interleukin-2 Interleukins Knock-in Mouse Knock-out Knockout Mice Liver Lymphocyte Lymphoid Cell Lymphoid Tissue Metabolic Modeling Modification Mouse Strains Mus Mutation Natural Killer Cells Non obese Nuclear Organ Paper Pathology Peyer&apos s Patches Phenotype Population Prevention RAG1 gene Reproduction Research Personnel SCID Mice Signal Transduction Systems Development T-Lymphocyte TSLP gene Testing Translational Research Transplantation Vaccination Vaccines Virus Replication Yellow Fever adaptive immune response adaptive immunity body system chemokine cytokine design diabetic drug discovery emerging pathogen enzyme deficiency hepatocyte engraftment human pathogen human stem cells humanized mouse improved knockout gene lymph nodes lymphoid organ lymphotoxin beta receptor member mouse genome mouse model paralogous gene preservation receptor response secondary lymphoid organ stem cell engraftment therapeutic development therapeutic evaluation therapeutically effective tool transcription factor tumor immunology vaccine discovery vaccine evaluation

项目摘要

Animal models are essential for studying biological processes underlying human health and diseases and developing safe and effective therapeutic approaches before human clinical trials. We dedicated our proposal to developing and characterizing new and significantly improved genetically modified animal models for human immune system establishment in mice. The conception of genetically engineered mice to engraft functional human immune systems opened a new horizon to study human-specific infections and associated multiorgan pathology. These models enable the successful engraftment of stem cells of non-fetal human tissue origin, including ex vivo engineered cells. Humanized mice are permissible to direct infection or challenges with wild- type human pathogens. Moreover, human cells isolated from experimental models became valuable for analyzing transcriptional and metabolic changes during infections and treatment. Thus, humanized mice have allowed researchers to address questions related to the treatment and prevention of important diseases like human immunodeficiency virus, hepatitis viruses (HIV/HBV/HDV/HCV), and newly emerging pathogens. Such models are directly applicable to study human health and diseases like human-specific infections, cancer immunology, transplantation of genetically modified human stem cells, and phenotypic characterization of various organ systems by omics approaches. We designed a new mouse background to avoid common cytokine gamma chain knockout and preserved secondary lymphoid organs for the efficient population with human immune cells. Nuclear factor interleukin-3 (Nfil3; also known as E4-binding protein 4, E4Bp4) transcription factor will be knocked out by CRISPR/Cas technology. By introducing human receptors and chemokines involved in the formation and growth of lymphoid tissues, we will improve the development of human adaptive immunity. To enable new strains of mice with the improved human immune system for the studies of human-specific hepatocytes infections, we will introduce fumarylacetoacetate hydrolase (Fah) gene knockout. Disruption of Fah gene on these new backgrounds will induce enzyme deficiency, currently regarded as the best model for human hepatocytes engraftment. Combining strain modifications will facilitate creating a dual humanized mouse model with immune system and liver to study human-specific infections, therapeutics development, and evaluation of vaccines. We will test our hypothesis by completing two specific aims: 1) to characterize the development and function of the human immune system in Nfil3/E4Bp4 knockout NOD/scid mice. Further improvement of human immune system functionality will be achieved by expressing the human lymphotoxin beta receptor, the chemokine CXCL13, and the thymic stromal lymphopoietin; 2) To disrupt Fah gene activity on NOD/scid-Nfil3-/- strain using CRISPR/Cas approaches. Advances in human immune system reproduction will fulfill increasing demands for developing improved animal models that are more predictable, accessible, and widely applicable for biomedical research.

动物模型对于研究人类健康和疾病的生物过程至关重要在人体临床试验之前开发安全有效的治疗方法。我们致力于我们的建议开发和表征新的和显着改进的人类转基因动物模型小鼠免疫系统的建立。基因工程小鼠的概念植入功能人类免疫系统为研究人类特异性感染和相关多器官开辟了新视野病理。这些模型能够成功植入非胎儿人体组织来源的干细胞，包括离体工程细胞。人源化小鼠可以直接感染或挑战野生型人类病原体。此外，从实验模型中分离出的人类细胞对于以下方面变得有价值：分析感染和治疗期间的转录和代谢变化。因此，人源化小鼠具有允许研究人员解决与治疗和预防重要疾病相关的问题，例如人类免疫缺陷病毒、肝炎病毒（HIV/HBV/HDV/HCV）和新出现的病原体。这样的模型直接适用于研究人类健康和疾病，如人类特异性感染、癌症免疫学、转基因人类干细胞移植以及表型表征通过组学方法研究各种器官系统。我们设计了新的小鼠背景以避免常见的细胞因子伽玛链敲除并保留次级淋巴器官是人类免疫细胞的有效群体。核因子白细胞介素3 （Nfil3；也称为E4结合蛋白4，E4Bp4）转录因子将被CRISPR/Cas敲除技术。通过引入参与淋巴形成和生长的人类受体和趋化因子组织，我们将改善人类适应性免疫的发展。为了使新的小鼠品系能够改善人体免疫系统，用于研究人类特异性肝细胞感染，我们将介绍富马酰乙酰乙酸水解酶（Fah）基因敲除。 Fah 基因在这些新背景下的破坏将诱导酶缺乏，目前被认为是人类肝细胞移植的最佳模型。组合菌株修饰将有助于创建具有免疫系统和肝脏的双重人源化小鼠模型以供研究人类特异性感染、治疗方法开发和疫苗评估。我们将通过以下方式检验我们的假设完成两个具体目标：1）表征人类免疫系统的发育和功能 Nfil3/E4Bp4 敲除 NOD/scid 小鼠。人体免疫系统功能的进一步提高通过表达人淋巴毒素β受体、趋化因子CXCL13和胸腺基质来实现淋巴细胞生成素； 2) 使用 CRISPR/Cas 方法破坏 NOD/scid-Nfil3-/- 菌株上的 Fah 基因活性。人类免疫系统繁殖的进步将满足开发改良动物日益增长的需求更可预测、更容易获得并且广泛适用于生物医学研究的模型。