GLAUCOMA THERAPY, CILIARY MUSCLE CONTRACTION AND TRABECULAR OUTFLOW

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 目的：研究产生一氧化氮(NO)的化合物对体外收缩或静息的睫状肌(CM)是否具有松弛作用。这是一种指标，表明这类化合物作为降低青光眼眼内压(IOP)的一种方法，是否有助于增强葡萄膜巩膜流出。研究化合物对体外心肌收缩/松弛及体内眼压的影响。利用器官培养中的猴眼前节细胞，研究基因治疗和其他分子对小梁流出的影响，这可能对青光眼的治疗也有重要意义。目的：探讨活体基因治疗对眼前节的影响。改变房水流出的蛋白质的过度表达可能导致眼压降低，这对青光眼治疗很重要。或者，眼压升高可能导致一种新的青光眼模型。 进展：在给予一氧化氮供体之前，使用鸟苷环化酶途径的抑制剂ODQ可阻断心肌对一氧化氮供体的松弛反应。没有化合物在CM需要松弛或收缩的治疗领域具有潜在价值，例如在青光眼治疗中。 猴子器官培养眼前段(MOCAS)系统正被用来确定药物治疗和基因治疗对小梁流出的影响，这可能随后被用于青光眼治疗以降低眼压。Cochlin蛋白的过度表达只在人类青光眼中升高，会增加眼压(IOP)并减少小梁流出。在培养液中加入牛血清白蛋白可增强转化生长β2(TGF？2)治疗后MOCAS患者的眼压升高反应。在转化生长因子2治疗过程中抑制Cochlin表达的研究将确定转化生长因子2诱导的眼压升高是否需要Cochlin诱导。 表达可能改变房水流出的基因的慢病毒载体已经被注射到活着的猴子眼睛的前段。转导C3转移酶(能够改变肌动蛋白细胞骨架)的一只眼不会改变眼压。眼部组织正在接受检查，以确认是否存在表达。另一种名为前列腺素合成酶的基因(预计会促进葡萄膜巩膜流出)被传递到5只猴子各一只眼睛的前段。注射后2周至5个月，5只眼中有3只眼的眼压有一过性下降。近2年来，在体内检测到绿色荧光蛋白的共表达。将检查组织中PGF合成酶的表达。 眼部局部涂抹维生素D可降低眼压。这种减少的机制正在调查中。 这项研究使用了WNPRC Research Services。 出版物： Lee E，Gabelt BT，Faralli JA，Peters DM，Brandt CR，Kaufman PL，Bhattacharya SK：Coch腺病毒在培养的人眼小梁细胞中的转基因表达及其对猴子器官培养的前节流出能力的影响。投资眼科VS科学，Epub出版前，2009年11月20日。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Objective: To determine whether compounds that generate nitric oxide (NO) can relax carbachol contracted or resting ciliary muscle (CM) in vitro. This is an indicator of whether this class of compounds may be useful in enhancing uveoscleral outflow as an approach for lowering intraocular pressure (IOP) in glaucoma. To determine the effects of compounds on in vitro CM contraction/relaxation, and on IOP in vivo. To utilize the monkey anterior segment in organ culture to investigate the effects of gene therapy and other molecules on trabecular outflow which may also be important for glaucoma therapy. To determine the effects of in vivo gene therapy to the anterior segment. Overexpression of proteins that alter aqueous humor outflow may lead to intraocular pressure reduction that is important for glaucoma therapy. Alternatively, elevation of intraocular pressure may lead to a new glaucoma model. Progress: The CM relaxation response to nitric oxide donors was blocked by pretreatment with ODQ, an inhibitor of the guanylate cyclase pathway, prior to administering a nitric oxide donor. NO compounds have potential value in therapeutic areas where relaxation or contraction of the CM is desirable, such as in the treatment of glaucoma. The monkey organ-cultured anterior segment (MOCAS) system is being utilized to determine the effects of pharmacotherapy and gene therapy on trabecular outflow which may subsequently be utilized for glaucoma therapy to decrease intraocular pressure. Overexpression of the protein cochlin, which is elevated only in human glaucoma, increases intraocular pressure (IOP) and decreases trabecular outflow. The IOP elevation response in MOCAS following treatment with transforming growth beta-2 (TGF¿2) is enhanced by including bovine serum albumin in the media. Studies to silence cochlin expression during TGF¿2 treatment will determine if cochlin induction is necessary for the TGF¿2-induced IOP elevation. Lentiviral vectors expressing genes that can potentially alter aqueous humor outflow have been injected into the anterior segment of living monkey eyes. Transduction of C3 transferase (capable of altering the actin cytoskeleton) in one eye did not alter the IOP. Ocular tissue is being examined to verify expression was present. Another gene, prostaglandin synthase (expected to enhance uveoscleral outflow), was delivered to the anterior segments of one eye of each of 5 monkeys. A transient reduction in IOP was detected in 3 of 5 eyes from 2 weeks to 5 months post-injection. Coexpression of green fluorescent protein was detectable in vivo for nearly 2 years. Tissues will be examined for PGFsynthase expression. Vitamin D applied topically to the eye lowers IOP. The mechanisms for this reduction are being investigated. This research used WNPRC Research Services. PUBLICATION: Lee E, Gabelt BT, Faralli JA, Peters DM, Brandt CR, Kaufman PL, Bhattacharya SK: COCH transgene expression by adenovirus in cultured human trabecular meshwork cells and its effect on outflow facility in monkey organ-cultured anterior segments. Invest Ophthalmol Vis Sci, Epub ahead of print, Nov 20, 2009.