ANTISENSE INHIBITION OF RETINAL GLUTAMATE METABOLISM

这个K 08项目的目的是测试目前关于谷氨酸在青光眼视网膜神经节细胞（RGC）死亡中的作用的两个主要假设之一。根据该假设，IOP升高的一个结果是RGC中谷氨酸代谢循环的紊乱。这种紊乱导致谷氨酸兴奋性毒性，其在青光眼的发病机制中具有主要作用。本项目的目标1和目标2测试该假设的第二部分;目标3测试第一部分。目的1是确定视网膜谷氨酸代谢周期紊乱对RGC功能和存活的影响。谷氨酸代谢酶、谷氨酸脱氢酶（GDH）和谷氨酰胺合成酶（GS）以及谷氨酸转运蛋白（GLAST）的反义寡核苷酸将用于抑制RGC中的谷氨酸代谢和摄取。将在组织培养中测定它们的作用，然后在玻璃体内给药大鼠后测定它们的作用，以评估RGC存活、活性和GDH的量。GS和GLAST、玻璃体内谷氨酸浓度和视网膜电图检查的视功能。目的2：检测谷氨酸拮抗剂和抗凋亡剂是否可以逆转反义寡核苷酸引起的视网膜谷氨酸循环紊乱的作用。目的3是确定在大鼠青光眼的慢性高眼压模型中是否存在类似于通过使用GDH、GS和GLAST的反义寡核苷酸诱导的谷氨酸代谢周期的变化。K 08候选人最近完成了青光眼领域的临床研究，并开始了在山终身教职的学术生涯。西奈医学院。虽然他之前确实有一些基础研究经验，但为了着手进行拟议的研究，他必须首先加强他在反义设计和药代动力学领域的技能，以及将用于测试动物和评估结果的一些特定方法。指导将由这些领域的资深科学家和专家提供。

The objective of this K08 project is to test one of the two major current hypotheses about the role of glutamate in glaucomatous retinal ganglion cell (RGC) death. According to this hypothesis one result of elevated IOP is a derangement of the glutamate metabolic cycle in RGCs. This derangement results in glutamate excitotoxicity that has a primary role in the pathogenesis of glaucoma. Aim 1 and 2 of this project test the second component of this hypothesis; aim 3 tests the first component. Aim 1 is to determine the effects of a deranged glutamate metabolic cycle in the retina on the RGC function and survival. Antisense oligonucleotides to the glutamate metabolizing enzymes, glutamate dehydrogenase (GDH) and glutamine synthetase (GS) and to glutamate transporters (GLAST) will be used to inhibit glutamate metabolism and uptake in RGCs. Their effects will be determined in tissue culture and then following intra-vitreal administration rats to assess RGC survival, activities and amounts of GDH. GS and GLAST, intra-vitreal glutamate concentrations, and visual function by electroretinography. Aim 2 tests whether glutamate antagonists and anti-apoptosis agents can reverse the effects of a deranged glutamate cycle in the retina caused by the antisense oligonucleotides. Aim 3 is to determine whether changes in the glutamate metabolic cycle similar to those induced by the use of antisense oligonucleotides to GDH, GS and GLAST are present in a chronically elevated IOP model of glaucoma in rats. The K08 candidate has recently completed a clinical fellowship in the field of glaucoma and has begun a tenure track academic career in Mt. Sinai School of Medicine. While he does have some prior fundamental research experience, in order to embark upon the proposed studies he must first strengthen his skills in the areas of antisense design and pharmacokinetics and in some specific methods that will be used in testing the animals and assessing the outcomes. Mentoring will be provided by senior scientists, experts in these areas.