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

基本信息

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
项目类别：
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

项目摘要

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）：治疗多发性硬化症（MS）的一个核心问题是如何促进中枢神经系统细胞的存活，因为疾病的进行性会导致轴突损伤、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。 AKT3 是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-/- 小鼠会敏感地发展为髓磷脂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)，这是一种具有 MS 方面的动物模型，与野生型 (WT) 小鼠相比，会导致更严重的疾病。病理分析表明，AKT3-/-小鼠的轴突损伤和脱髓鞘发生率比患有慢性疾病的WT小鼠更高。骨髓移植实验表明，免疫系统和中枢神经系统内的细胞促成了这种效应。该提案的目的是确定导致在缺乏 AKT3 的小鼠中观察到的更严重表型的细胞。为此，将产生 AKT3-floxed 小鼠的种系传播。然后将小鼠与 FLP 小鼠杂交以删除 FRT 位点并生成 AKT3-floxed 条件小鼠。然后将每个-Cre品系与AKT3floxed条件小鼠杂交以产生F1。将产生在选定的免疫细胞、神经元和少突胶质细胞中删除 AKT3 的 AKT3 条件敲除 (KO) 小鼠。将评估这些小鼠和 floxed 小鼠。一旦我们确信获得了 KO，我们将在 MOG 诱导的 EAE 过程中检查这些小鼠。这些研究将扩大我们对 AKT3 对免疫活性细胞功能的贡献以及 AKT3 在 CNS 中的作用的理解。我们的长期目标是表征 AKT3 在 CNS 和免疫细胞中的作用，以便治疗方法可以集中于受 AKT3 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。