The Role of CaMKII in Cell Death and Survival Responses of Retinal Ganglion Cells

CaMKII 在视网膜神经节细胞死亡和存活反应中的作用

• 批准号: 7625898
• 负责人: NIGEL G.F. COOPER
• 金额: $ 35.5万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7625898
• 关键词: Acute; Affect; Agonist; Animal Model; Anterior Ischemic Optic Neuropathy; Blindness; Brain-Derived Neurotrophic Factor; Calcium/calmodulin-dependent protein kinase; Caspase; Cell Death; Cell Line; Cell Survival; Cells; Cessation of life; Clinical; Data; Development; Diabetic Retinopathy; Disease; Equilibrium; Event; Eye diseases; Ganglion Cell Layer; Gene Targeting; Genes; Genetic Transcription; Glaucoma; Glutamate Receptor; Glutamates; Goals; In Vitro; Injection of therapeutic agent; Intraperitoneal Injections; Ischemia; Knowledge; Laboratories; Link; Mediating; Methods; Modeling; Nuclear; Pathway interactions; Patients; Peptides; Phosphotransferases; Play; Process; Protein Isoforms; RNA Interference; RNA Splicing; Rattus; Regulator Genes; Reperfusion Therapy; Research; Retina; Retinal; Retinal Diseases; Retinal Ganglion Cells; Role; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Site; Testing; Therapeutic Intervention; Time; Transcript; Transduction Gene; Visual impairment; autocrine; calmodulin-dependent protein kinase II; caspase-3; inhibitor/antagonist; insight; neuroprotection; paracrine; prospective; public health relevance; response; retinal ischemia; retinal neuron; transcription factor; Acute; Affect; Agonist; Animal Model; Anterior Ischemic Optic Neuropathy; Blindness; Brain-Derived Neurotrophic Factor; Calcium/calmodulin-dependent protein kinase; Caspase; Cell Death; Cell Line; Cell Survival; Cells; Cessation of life; Clinical; Data; Development; Diabetic Retinopathy; Disease; Equilibrium; Event; Eye diseases; Ganglion Cell Layer; Gene Targeting; Genes; Genetic Transcription; Glaucoma; Glutamate Receptor; Glutamates; Goals; In Vitro; Injection of therapeutic agent; Intraperitoneal Injections; Ischemia; Knowledge; Laboratories; Link; Mediating; Methods; Modeling; Nuclear; Pathway interactions; Patients; Peptides; Phosphotransferases; Play; Process; Protein Isoforms; RNA Interference; RNA Splicing; Rattus; Regulator Genes; Reperfusion Therapy; Research; Retina; Retinal; Retinal Diseases; Retinal Ganglion Cells; Role; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Site; Testing; Therapeutic Intervention; Time; Transcript; Transduction Gene; Visual impairment; autocrine; calmodulin-dependent protein kinase II; caspase-3; inhibitor/antagonist; insight; neuroprotection; paracrine; prospective; public health relevance; response; retinal ischemia; retinal neuron; transcription factor

DESCRIPTION (provided by applicant): The signal transduction and gene regulatory events which occur downstream of the precipitating causes of retinal diseases is largely unknown. Yet these downstream sites would be excellent targets for interventional therapies. The long term goal of the proposed research is to define the role of calmodulin dependent protein kinase II (CaMKII) which is likely downstream of the initiating events in eye diseases like retinal ischemia. CaMKII is upstream of the cell death and survival pathways in retinal ganglion cells (RGCs). We know that when challenged with glutamate receptor agonists: 1) retinal cytoplasmic CaMKII1 becomes autophosphorylated; 2) there is an increase in the alternatively spliced nuclear transcript, CaMKII1B; 3) caspase-3 becomes activated and 4) cells in the inner nuclear and ganglion cell layers die. We have demonstrated that the specific inhibitor of CaMKII, autocamtide inhibitory peptide (AIP), blocks these changes (1, 3 and 4) and protects all cells from cell death. Pilot data with the aid of a cell line indicates that the AIP promotes the secretion of brain derived neurotrophic factor (BDNF) which may aid the survival of challenged cells. We also know that an elevated level of CaMKII1B (2) is associated with enhanced transcription of BDNF. Since glutamate is implicated in retinal ischemia and CaMKII1 is also activated in this condition, we hypothesize that CaMKII1 plays an important role in pathways involved in the death and survival responses of RGCs during retinal ischemia. This will be tested with methods, which include RNA- interference for the CaMKII1B transcript. We will use a rat retinal ischemia-reperfusion model, as well as primary cultured rat RGCs, and ex vivo retinal explants. Aim 1 will determine the time frame in which AIP protects RGCs during/after ischemia. Aim 2 will explore the role of cytoplasmic CaMKII in regulating caspase-3 activation, NF:B translocation and BDNF release in RGC/retinas. Aim 3 will explore the role of nuclear CaMKII1 in cell death/survival and the underlying mechanisms. The results of these studies will provide important insights into the signal transduction pathways and gene regulatory networks that control death and survival responses of RGCs which will be of relevance to retinal ischemia. PUBLIC HEALTH RELEVANCE: Retinal ischemia, a common clinical entity that has been widely studied because of its proposed role in, for example, retinal and choroidal vessel occlusion, anterior ischemic optic neuropathy, glaucoma, and diabetic retinopathy, etc, remains a common cause of blindness in the developed world due to relatively ineffective treatment. We propose to explore the signal transduction pathways linked to gene regulatory networks which control cell death and survival processes in rat retinal ganglion cells, whose death is a major contributing factor for visual impairment in these diseases. Knowledge of such pathways and networks is important for the development of practical therapeutic interventions.

描述（由申请人提供）：在视网膜疾病的降水原因下游发生的信号转导和基因调节事件在很大程度上尚不清楚。然而，这些下游部位将是介入疗法的绝佳目标。拟议的研究的长期目标是定义钙调蛋白依赖性蛋白激酶II（CAMKII）的作用，这很可能是视网膜缺血等眼部疾病中引发事件的下游。 CAMKII是视网膜神经节细胞（RGC）中细胞死亡和存活途径上游的上游。我们知道，当受到谷氨酸受体激动剂的挑战时：1）视网膜胞质Camkii1被自磷酸化； 2）剪接的核转录本camkii1b有所增加； 3）caspase-3被激活，4）内部核和神经节细胞层中的细胞死亡。我们已经证明，CAMKII，自X型抑制肽（AIP）的特异性抑制剂阻止了这些变化（1、3和4），并保护所有细胞免受细胞死亡的影响。借助细胞系的飞行员数据表明​​，AIP促进了脑衍生的神经营养因子（BDNF）的分泌，这可能有助于挑战细胞的存活。我们还知道，CAMKII1b（2）的水平升高与BDNF的转录增强有关。由于谷氨酸与视网膜缺血有关，并且在这种情况下也激活了CAMKII1，因此我们假设CaMKII1在视网膜缺血期间RGC的死亡和生存反应涉及的途径中起重要作用。这将通过方法进行测试，其中包括对CAMKII1B转录本的RNA干扰。我们将使用大鼠视网膜缺血再灌注模型以及原代培养的大鼠RGC和离体视网膜外植体。 AIM 1将确定AIP在缺血期间/之后保护RGC的时间范围。 AIM 2将探讨细胞质CAMKII在调节caspase-3激活，NF：B易位和BDNF释放中的作用。 AIM 3将探讨核CAMKII1在细胞死亡/生存和潜在机制中的作用。这些研究的结果将提供对控制与视网膜缺血有关的RGC的死亡和生存反应的信号转导途径和基因调节网络的重要见解。公共卫生相关性：视网膜缺血，这是一个普通的临床实体，由于其在例如视网膜和脉络膜血管闭塞，前缺血性神经性神经病，血糖瘤和糖尿病视网膜病等方面的作用而受到广泛研究，这仍然是发展世界中盲目的常见原因。我们建议探索与基因调节网络相关的信号转导途径，这些途径控制大鼠视网膜神经节细胞中的细胞死亡和存活过程，后者的死亡是这些疾病视觉障碍的主要因素。对这种途径和网络的了解对于开发实际治疗干预措施很重要。