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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：
10548100
负责人：
Santhi Gorantla
金额：
$ 23.03万
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/10548100
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 physiology Cells Clinical Trials Communities Conceptions Cytokine Signaling Development Disease Distant Engineering Engraftment 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 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 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 base body system chemokine cytokine design diabetic drug discovery emerging pathogen enzyme deficiency hepatocyte engraftment human pathogen human stem cells humanized mouse immune system function improved improved functioning 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。我们的提案致力于为人类开发和表征新的和显著改进的转基因动物模型小鼠免疫系统建立。功能性移植基因工程小鼠的构想人类免疫系统为研究人类特异性感染和相关的多器官病理这些模型能够成功植入非胎儿人组织来源的干细胞，包括离体工程细胞。允许人源化小鼠直接感染或攻击野生型- 类型人类病原体。此外，从实验模型中分离出的人类细胞对于分析感染和治疗期间的转录和代谢变化。因此，人源化小鼠具有使研究人员能够解决与治疗和预防重要疾病有关的问题，人类免疫缺陷病毒、肝炎病毒（HIV/HBV/HDV/HCV）和新出现的病原体。等模型直接适用于研究人类健康和疾病，如人类特有的感染，癌症，免疫学、遗传修饰的人干细胞移植和通过组学方法研究不同的器官系统。我们设计了一种新的小鼠背景，以避免常见的细胞因子γ链敲除，并保留二次淋巴器官的有效人口与人类免疫细胞。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.通过引入参与淋巴细胞形成和生长的人受体和趋化因子，组织，我们将提高人类适应性免疫的发展。为了使新品系的小鼠具有为了研究人类特异性肝细胞感染，我们将介绍延胡索酰乙酰乙酸水解酶（Fah）基因敲除。Fah基因在这些新背景下的破坏将诱导酶缺乏，目前被认为是人肝细胞移植的最佳模型。结合品系修饰将有助于建立具有免疫系统和肝脏的双重人源化小鼠模型以进行研究人类特异性感染、治疗开发和疫苗评估。我们将测试我们的假设，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。人类免疫系统功能的进一步改善将是通过表达人光敏素β受体、趋化因子CXCL 13和胸腺基质细胞， 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展将满足开发改良动物的日益增长的需求模型更可预测，更容易获得，并广泛适用于生物医学研究。