Gene Expression Analysis in Microcaptured Retinal Cells

描述(申请人提供)：视网膜退行性疾病是全球范围内导致视力残疾和失明的主要原因。例如，老年性黄斑变性(AMD)是西方世界老年人失明的主要原因。目前的治疗方法对改变视网膜退化造成的不可阻挡的视力丧失几乎没有什么作用。一些研究表明，眼内注射脑源性神经营养因子(BDNF)、睫状神经营养因子(CNTF)或碱性成纤维细胞生长因子-2(FGF2)等因子，可以减缓特定突变或持续光照引起的光感受器细胞死亡。然而，这些发现的临床用途可能是有限的，因为救援效果是部分和短暂的，而且一些因素似乎有不想要的副作用。为了最大化益处和最小化副作用，阐明生存因子延缓视网膜退化的机制似乎是必要的。我们实验室最近的研究表明，CNTF、BDNF和FGF2并不直接作用于光感受器；相反，它们似乎是通过其他细胞间接作用的，很可能是M勒氏细胞。在这些观察的基础上，我们建议研究神经营养因子在小鼠体内引发的分子变化。勒氏细胞。这些研究涉及两种互补且要求苛刻的最先进技术的组合使用：从单个细胞产生cDNA，以及使用定制设计的视网膜cDNA微阵列进行分析。然后，我们将确定这些变化中的哪些对感光细胞的存活是重要的。识别这些分子的潜在影响是显而易见的，因为它们可能为这些毁灭性疾病的治疗提供新的途径。

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.