Enhancement of Human Immune System Development in Mouse Models

动物模型对于研究潜在的人类健康和疾病的生物过程是必不可少的 在人类临床试验之前开发安全有效的治疗方法。我们将我们的建议献给 为人类开发和表征新的和显著改进的转基因动物模型 小鼠免疫系统的建立。基因工程小鼠移植功能的构想 人类免疫系统开辟了研究人类特有感染及其相关多器官的新视野 病理学。这些模型使非胎儿人体组织来源的干细胞能够成功植入， 包括体外工程细胞。人源化的小鼠可以直接感染或挑战野生- 人类病原体的类型。此外，从实验模型中分离出的人类细胞对 分析感染和治疗期间的转录和代谢变化。因此，人源化的小鼠 允许研究人员解决与治疗和预防重要疾病有关的问题，如 人类免疫缺陷病毒、肝炎病毒(艾滋病毒/乙肝病毒/HDV/丙型肝炎病毒)和新出现的病原体。是这样的 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，转基因人类干细胞的移植和表型特征 通过组学方法研究各种器官系统。 我们设计了一种新的小鼠背景，避免了常见的细胞因子伽马链被敲除和保存 次级淋巴器官为有效人群的人体免疫细胞。核因子白介素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样细胞形成和生长的人类受体和趋化因子 组织方面，我们将提高人类适应性免疫的发展。为了使新品系的小鼠能够 针对人类免疫系统改善对人类特异性肝细胞感染的研究，我们将介绍 富马酸乙酰乙酸酯水解酶(Fah)基因敲除。在这些新背景上破坏Fah基因将 诱导酶缺乏症，目前被认为是最好的人肝细胞植入模型。组合 品系改良将有助于建立免疫系统和肝脏双重人源化小鼠模型进行研究 人类特有的感染、治疗方法的开发和疫苗的评估。我们将通过以下方式验证我们的假设 完成两个具体目标：1)描述人类免疫系统的发育和功能 NFIL3/E4Bp4基因敲除NOD/SCID小鼠。人类免疫系统功能的进一步改善将是 通过表达人类淋巴毒素β受体、趋化因子CXCL13和胸腺基质来实现 2)用CRISPR/Cas方法阻断NOD/SCID-Nfil3-/-株的Fah基因活性。 人类免疫系统繁殖的进展将满足发展改良动物的日益增长的需求 更可预测、更容易获得、更广泛适用于生物医学研究的模型。

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.