Pressure Regulation of Human Trabecular Meshwork Genes

描述(申请人提供)：眼内的生理压力由小梁网(商标)组织提供的阻力机制维持。在大多数情况下，长期的压力升高会导致视网膜神经节细胞变性、视神经损伤和失明(青光眼)。我们的长期目标是寻找在调节眼压(LOP)中起重要作用的TM基因/机制。在这个项目的第一个周期中，我们使用宏阵列和基因芯片技术来获得压力升高的眼睛和它们的正常眼压对照之间的差异表达谱。一些已识别的基因编码新的、以前未描述的TM功能，这些功能似乎是从骨骼和血管系统招募来的。与其继续识别更多的基因，我们建议利用所获得的信息来研究一些新的机制和LOP参数。 我们的工作假设是LOP的调节在一定程度上是由TM基因的协调表达决定的。根据我们的发现，我们现在假设，阻止TM的细胞外基质(ECM)钙化可能是调节LOP的关键机制。我们进一步假设，差异表达的具有高倍数变化值的基因可能是识别压力反应的启动子元件的理想候选。最后，我们假设每个个体通过诱导不同程度的压力反应基因，从而改变它们的相对丰度，从而对压力做出不同的反应。由于压力反应基因编码参与不同功能的蛋白质，这种相对丰度可能会影响个体的压力阈值，并导致他/她患上这种疾病的风险。我们将通过使用人类TM细胞和灌流器官培养、重组正义和siRNA腺病毒、构象抗体、报告基因和定量PCR来验证这些假设。

DESCRIPTION (provided by applicant): Physiological pressure inside the eye is maintained by a resistance mechanism provided by the trabecular meshwork (TM) tissue. In most cases, prolonged elevated pressure leads to retinal ganglion cell degeneration, optic nerve damage and blindness (Glaucoma). Our long-term goal is to search for TM genes/mechanisms playing an essential role in regulation of intraocular pressure (lOP). During the first cycle of this project we used macroarray and GeneChip technology to obtain differential expression profiles between eyes subjected to elevated pressure and their paired normotense controls. Some of the identified genes encode new, previously undescribed TM functions, which appear recruited from bone and vascular systems. Rather than continue identifying more genes, with the information obtained we propose to investigate some of the new mechanisms and lOP parameters. Our working hypothesis is that regulation of lOP is governed in part by the coordinated expression of TM genes. Based on our findings, we now hypothesize that preventing calcification of the extracellular matrix (ECM) of the TM might be a key mechanism contributing to the regulation of lOP. We further hypothesize that genes differentially expressed with high fold change values might serve as ideal candidates to identify promoter elements responding to pressure. Finally, we hypothesize that each individual responds differently to pressure by inducing to different extent pressure-responding genes, thus changing their relative abundance. Because pressure-responding genes encode proteins involved in different functions, this relative abundance could influence an individual pressure threshold and contribute to his/her risk of developing of the disease. We will test these hypotheses by using human TM cell and perfused organ cultures, recombinant sense and siRNA adenoviruses, conformational antibodies, reporter genes, and quantitative PCR.