Generation of floxed-AKT3 mice to study the role in neuroinflammatory diseases

生成 floxed-AKT3 小鼠以研究其在神经炎症疾病中的作用

• 批准号: 8695552
• 负责人: BRIDGET SHAFIT-ZAGARDO
• 金额: $ 6.85万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8695552
• 关键词: AKT3 gene; Abnormal Cell; Acute; Affect; Age; Animal Model; Biological Assay; Bone Marrow; Brain; Cell Death; Cell Survival; Cells; Cellular Morphology; Chronic Disease; Data; Defect; Demyelinating Diseases; Demyelinations; Disease; Disease model; Effector Cell; Experimental Autoimmune Encephalomyelitis; Family member; Future; Generations; Goals; Homeostasis; Human; ITGAM gene; Immune; Immune system; Immunocompetent; Individual; Inflammation; Inflammatory; Injury; Interferons; Interleukin-17; Interleukin-2; Knock-out; Knockout Mice; Maintenance; Messenger RNA; Modeling; Movement; Multiple Sclerosis; Mus; Nature; Neuraxis; Neurons; Oligodendroglia; Pathway interactions; Phenotype; Prevalence; Production; Protein-Serine-Threonine Kinases; Proto-Oncogene Proteins c-akt; Public Health; Regulatory T-Lymphocyte; Relative (related person); Research; Role; Severities; Signal Transduction; Signaling Molecule; Site; Spinal Cord; Synapsins; T-Lymphocyte; Technology; Therapeutic; Tissues; cell growth; cell type; cytokine; improved; insight; mouse model; myelination; oligodendrocyte-myelin glycoprotein; public health relevance; recombinase; remyelination; research study; transmission process

DESCRIPTION (provided by applicant): A central issue for the treatment of multiple sclerosis (MS) is how to promote the survival of cells that reside in the CNS since the progressive nature of the disease results in axonal damage, neuronal cell loss, oligodendrocyte cell death, and permanent demyelination. AKT serine-threonine kinase family members are important regulators of cell growth, proliferation, and cell survival. AKT3, the major form expressed in the brain, is found in neurons and oligodendrocytes, but is also expressed in the immune system. We determined that AKT3-/- mice sensitized to develop myelin oligodendrocyte glycoprotein (MOG)-induced experimental autoimmune encephalomyelitis (EAE), an animal model with aspects of MS, resulted in more severe disease than in wildtype (WT) mice. Pathological analysis showed that AKT3-/- mice had increased prevalence of axonal damage and demyelination than WT mice with chronic disease. Bone marrow transfer experiments indicated that cells within both the immune system and the CNS contributed to this effect. The objective of this proposal is to define the cells contributing to the more severe phenotype observed in mice lacking AKT3. To this end, germline transmission of AKT3-floxed mice will be generated. Mice will then be crossed with FLP mice to delete the FRT site and generate AKT3-floxed conditional mice. Then each -Cre strain will be crossed with the AKT3floxed conditional mice to generate F1. AKT3 conditional knock-out (KO) mice with deletion of AKT3 in select immune cells, in neurons, and in oligodendrocytes will be generated. These mice and the floxed mice will be evaluated. Once we are assured that the KO is obtained we will examine these mice during MOG-induced EAE. These studies will expand our understanding of the contribution of AKT3 to the function of immunocompetent cells versus AKT3's role within the CNS. Our long term goal is to characterize the role of AKT3 in the CNS and in immune cells so that therapeutic approaches might focus on signaling molecules affected by the AKT3 pathway, and provide protection against CNS inflammation and tissue destruction found in MS.

描述（由申请人提供）：治疗多发性硬化（MS）的中心问题是如何促进驻留在CNS中的细胞的存活，因为该疾病的进行性导致轴突损伤、神经元细胞损失、少突胶质细胞死亡和永久性脱髓鞘。AKT丝氨酸-苏氨酸激酶家族成员是细胞生长、增殖和细胞存活的重要调节因子。AKT 3是大脑中表达的主要形式，在神经元和少突胶质细胞中发现，但也在免疫系统中表达。我们确定，AKT 3-/-小鼠致敏发展髓鞘少突胶质细胞糖蛋白（MOG）诱导的实验性自身免疫性脑脊髓炎（EAE），MS方面的动物模型，导致更严重的疾病比野生型（WT）小鼠。病理学分析显示，AKT 3-/-小鼠的轴突损伤和脱髓鞘的患病率比患有慢性疾病的WT小鼠增加。骨髓移植实验表明，免疫系统和中枢神经系统内的细胞都有助于这种效果。本提案的目的是确定导致在缺乏AKT 3的小鼠中观察到的更严重表型的细胞。为此，将产生AKT 3-floxed小鼠的种系传递。然后将小鼠与FLP小鼠杂交以删除FRT位点并产生AKT 3-floxed条件小鼠。然后将每个-Cre品系与AKT 3floxed条件小鼠杂交以产生F1。将产生在选定免疫细胞、神经元和少突胶质细胞中AKT 3缺失的AKT 3条件性敲除（KO）小鼠。将对这些小鼠和floxed小鼠进行评价。一旦我们确信获得KO，我们将在MOG诱导的EAE期间检查这些小鼠。这些研究将扩大我们对AKT 3对免疫活性细胞功能的贡献与AKT 3在CNS中的作用的理解。我们的长期目标是表征AKT 3在CNS和免疫细胞中的作用，以便治疗方法可以集中于受AKT 3通路影响的信号分子，并提供针对MS中发现的CNS炎症和组织破坏的保护。