Development of Microglia Knockout Mouse

小胶质细胞敲除小鼠的研制

• 批准号: 10040196
• 负责人: Bogoljub Ciric
• 金额: $ 15.6万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10040196
• 关键词: Acquired Immunodeficiency Syndrome; Address; Adrenoleukodystrophy; Adult; Age; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Animal Model; Biology; Blood - brain barrier anatomy; Bone Marrow; Brain Diseases; Cause of Death; Cells; Central Nervous System Infections; Communicable Diseases; Data Analyses; Dependence; Development; Discipline; Disease; Disease model; Encephalitis; Ensure; Enterobacteria phage P1 Cre recombinase; Experimental Autoimmune Encephalomyelitis; Experimental Models; Exposure to; Funding; Generations; Genes; Genetic Transcription; Health; Homeostasis; Huntington Disease; Immune; Inflammation; Intervention; Knock-out; Knockout Mice; Longevity; LoxP-flanked allele; Macrophage Colony-Stimulating Factor Receptor; Maintenance; Malignant Neoplasms; Malignant neoplasm of brain; Microglia; Modeling; Mononuclear; Multiple Sclerosis; Mus; Myeloid Cells; Neuraxis; Outcome; Parkinson Disease; Partner in relationship; Phagocytes; Phenotype; Physiological Processes; Play; Receptor Gene; Receptor Signaling; Research; Research Personnel; Rett Syndrome; Role; Speed; Tamoxifen; The Jackson Laboratory; Time; TimeLine; Transgenic Mice; Transgenic Organisms; astrogliosis; base; bone cell; experimental study; macrophage; mouse model; nervous system development; novel; painful neuropathy; progenitor; progenitor system; targeted treatment; tool; transcription factor; treatment duration

SUMMARY Microglia are central nervous system (CNS) resident macrophages involved in maintaining CNS homeostasis. Importantly, they are also involved in a number of diseases, including Alzheimer’s disease, Huntington’s disease, amyotrophic lateral sclerosis, Parkinson’s disease, multiple sclerosis, infectious diseases and cancer. Despite their importance in health and disease, researchers still lack an experimental model that would enable simple and permanent depletion of microglia to study their functions and their potential as targets for therapy. The principal difficulty in studying microglia is their extensive similarity to other macrophages, which precludes depletion of microglia without concomitant depletion of macrophages. Making use of recent advances in microglia biology, we propose to characterize a transgenic mouse model, termed here Mg-ipKO that enables inducible, permanent, and highly selective depletion of microglia only. The depletion is based on expression of Sall1, a transcription factor expressed in microglia but not in other macrophages/myeloid cells. Sall1-driven expression of Cre recombinase will lead to knockout of CSF-1 receptor (CSF-1R) in microglia. Given that microglia survival is absolutely dependent on CSF-1R, its knockout will lead to depletion of microglia. The depletion will be tamoxifen-inducible and permanent. A similar concept for short- term microglia depletion by Sall1-driven expression of Cre to knockout CSF-1R has recently been proven successful, supporting the feasibility of the approach that we propose. We have recently generated novel Sall1-FRT-Cre transgenic mouse line, and we are currently crossing these mice with two other transgenic mouse lines (obtained from The Jackson Laboratory) to generate final triple transgenic Mg-ipKO mice with the capacity for inducible microglia depletion. We anticipate that we will have generated Mg-ipKO mice by the time funding for this project may start, allowing for an immediate start of experiments with these mice. We propose the specific aim: To characterize microglia depletion and its effects in Mg-ipKO mice. To determine the usefulness of the model, we will induce microglia depletion in adult Mg-ipKO mice by tamoxifen treatment. We will then determine the extent of the depletion and optimize tamoxifen treatment if necessary. Furthermore, we will characterize potential consequences of the depletion on CNS homeostasis (blood-brain barrier integrity, CNS inflammation, astrogliosis) and determine if, over time, the CNS repopulates with microglia that originate from either CNS progenitors, or with macrophages of bone marrow origin. We expect that by the completion of this proposed research we will have developed and characterized an advanced transgenic mouse model for the depletion of microglia that overcomes the limitations of existing approaches and provides a highly useful research tool for multiple biomedical disciplines.

概括 小胶质细胞是中心神经系统（CNS）驻留巨噬细胞，涉及维持中枢神经系统稳态。 重要的是，他们还参与了多种疾病，包括阿尔茨海默氏病，亨廷顿氏病， 肌萎缩性侧面硬化症，帕金森氏病，多种扇形，传染病和癌症。 研究人员在健康和疾病中的重要性仍然缺乏实验模型模型模型模型模型，使简单地简单。 和小胶质细胞的永久部署，以研究功能和作为治疗靶标的潜力 研究小胶质细胞的主要差异是它们与其他巨噬细胞的广泛相似性，这排除了 小胶质细胞的部署而无需伴随巨噬细胞的部署。 利用小胶质细胞生物学的最新进展，我们建议表征转基因小鼠 模型，在这里称为MG-IPKO，仅对小胶质细胞诱导，永久和高度选择性剥夺。 部署基于SALL1的表达，Sall1是一种在小胶质细胞中表达的转录因子，但在其他 巨噬细胞/髓样细胞。 （CSF-1R）在小胶质细胞中。 小胶质细胞的部署。 通过SALL1驱动的CRE表达到术语小胶质细胞部署到 成功，支持我们提出的方法的可行性。 我们最近生成了新颖的SALL1-FRT-CRE转基因小鼠系，我们目前正在越过 这些带有另外两种转基因小鼠系（从杰克逊实验室获得的小鼠）生成最终三重的小鼠 转基因MG-IPKO小鼠具有诱导小胶质细胞的能力。 根据该项目的资金，生成的MG-IPKO小鼠可能会​​开始 对小鼠进行实验。 我们提出了具体目的：表征小胶质细胞耗竭，是MG-IPKO小鼠的作用。 为了确定模型的有用性，我们将通过他莫昔芬诱导小胶质细胞depko小鼠 治疗。 此外，我们将表征CNS稳态（血液）耗竭的潜在共识 屏障完整性，CNS炎症，星形胶质细胞增多症），并确定中枢神经系统是否与小胶质细胞重盘 这起源于中枢神经系统祖细胞或骨髓来源的巨噬细胞。 我们期望通过这项支撑研究的组成，我们将开发和表征 耗尽的高级转基因鼠标模型，该模型克服了现有的限制 方法并为多个生物医学学科提供了高度USOFUL研究工具。