Gene Expression Analysis in Microcaptured Retinal Cells

描述（由申请人提供）：视网膜退行性疾病是世界范围内视力残疾和失明的主要原因。例如，年龄相关性黄斑变性（AMD）是西方世界老年人失明的主要原因。目前的治疗方法几乎无法改变由于视网膜变性而导致的不可避免的视力丧失。几项研究表明，眼内注射脑源性神经营养因子（BDNF）、睫状体神经营养因子（CNTF）或碱性成纤维细胞生长因子-2 （FGF2）等因子，可减缓由特定突变或暴露于恒定光引起的光受体细胞死亡。然而，这些发现的临床用途可能是有限的，因为抢救效果是局部的和短暂的，而且一些因素似乎有不想要的副作用。阐明存活因子延缓视网膜变性的机制似乎是必要的，以便最大限度地提高获益和减少副作用。我们实验室最近的研究表明，CNTF、BDNF和FGF2不直接作用于光感受器；相反，它们似乎通过其他细胞间接起作用，最有可能的是M
ller细胞。基于这些观察结果，我们建议研究神经营养因子在M
ller细胞中引发的分子变化。这些研究涉及两种互补且要求最高的技术的结合使用：从单个细胞中产生cDNA，以及使用定制设计的视网膜cDNA微阵列进行分析。然后，我们将确定哪些变化对光感受器的存活是重要的。识别这些分子的潜在影响是显而易见的，因为它们可以为治疗这些毁灭性疾病提供新的途径。

DESCRIPTION (provided by applicant): Retinal degenerative diseases are a major cause of visual disability and blindness worldwide. Age-related macular degeneration (AMD), for example, is the leading cause of blindness in the elderly in the Western world. Current treatments do little to alter the inexorable loss of vision due to retinal degenerations. Several studies have shown that intraocular injection of factors such as brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), or basic fibroblast growth factor-2 (FGF2), slows photoreceptor cell death caused by specific mutations or exposure to constant light. However, the clinical usefulness of these findings may be limited, because rescue effects are partial and transient, and some factors appear to have unwanted side effects. Elucidation of the mechanism by which survival factors delay retinal degenerations appears necessary in order to maximize benefits and minimize side effects. Recent studies from our laboratories have suggested that CNTF, BDNF and FGF2 do not act directly on photoreceptors; rather, they appear to act indirectly through other cells, most likely M
ller cells. Based on these observations, we propose to investigate the molecular changes triggered by neurotrophic factors in M
ller cells. The studies involve the combined use of two complementary and demanding state-of-the-art techniques: the generation of cDNA from individual cells, and their analysis using custom designed retinal cDNA microarrays. We will then establish which of these changes are important for photoreceptor survival. The potential impact of the identification of these molecules is clear, since they could offer new avenues for the treatment of these devastating diseases.