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: 9131865
• 负责人: BRIDGET SHAFIT-ZAGARDO
• 金额: $ 1.5万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE, INC
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9131865
• 关键词: Generation; floxed; AKT3; mice; study

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 AKT3floxed 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 will be evaluated and once assured that the conditional KO is obtained we will apply for additional funding to 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）：多发性硬化症（MS）治疗的核心问题是如何促进中枢神经系统中细胞的存活，因为该疾病的进行性会导致轴突损伤、神经元细胞丢失、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。AKT3是在大脑中表达的主要形式，存在于神经元和少突胶质细胞中，但也在免疫系统中表达。我们确定AKT3-/-小鼠对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）致敏，这是一种具有MS方面的动物模型，导致比野生型（WT）小鼠更严重的疾病。病理分析显示，与患有慢性疾病的WT小鼠相比，AKT3-/-小鼠轴突损伤和脱髓鞘的发生率增加。骨髓移植实验表明，免疫系统和中枢神经系统中的细胞都参与了这种作用。本提案的目的是确定在缺乏AKT3的小鼠中观察到的导致更严重表型的细胞。为此，将产生AKT3-floxed小鼠的种系传播。然后将小鼠与FLP小鼠杂交，删除FRT位点，产生AKT3floxed条件小鼠。然后将每个-Cre菌株与AKT3floxed条件小鼠杂交产生F1。将产生AKT3条件敲除（KO）小鼠，在选定的免疫细胞、神经元和少突胶质细胞中缺失AKT3。这些小鼠将被评估，一旦确定获得条件KO，我们将申请额外的资金来检查这些小鼠在mog诱导的EAE期间。这些研究将扩大我们对AKT3对免疫活性细胞功能的贡献与AKT3在中枢神经系统中的作用的理解。我们的长期目标是表征AKT3在中枢神经系统和免疫细胞中的作用，以便治疗方法可能集中在受AKT3途径影响的信号分子上，并为MS中发现的中枢神经系统炎症和组织破坏提供保护。