GLAUCOMA THERAPY, CILIARY MUSCLE CONTRACTION AND TRABECULAR OUTFLOW

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 中心，不一定是研究者的机构。 目的：研究产生一氧化氮（NO）的化合物对卡巴胆碱收缩或静息状态下的离体睫状肌（CM）的舒张作用。这是这类化合物是否可用于增强葡萄膜巩膜流出作为降低青光眼眼内压（IOP）的方法的指标。确定化合物对体外CM收缩/舒张和体内IOP的影响。利用猴眼前节器官培养，研究基因治疗和其他分子对小梁流出量的影响，这对青光眼的治疗也有重要意义。目的：探讨眼前节基因治疗的效果。改变房水流出的蛋白质的过表达可能导致眼内压降低，这对于青光眼治疗是重要的。 或者，眼内压升高可能导致新的青光眼模型。 进度：CM松弛反应，一氧化氮供体被阻断预处理与ODQ，鸟苷酸环化酶途径的抑制剂，然后给予一氧化氮供体。NO化合物在需要CM松弛或收缩的治疗领域中具有潜在价值，例如在青光眼的治疗中。 猴器官培养眼前节（MOCAS）系统被用来确定药物治疗和基因治疗对小梁流出的影响，随后可用于青光眼治疗，以降低眼内压。 仅在人青光眼中升高的蛋白质cochlin的过度表达增加眼内压（IOP）并减少小梁流出。 MOCAS中转化生长β 2（TGF β 2）治疗后的IOP升高反应通过在培养基中加入牛血清白蛋白而增强。 在TGF β 2治疗期间沉默cochlin表达的研究将确定cochlin诱导是否是TGF β 2诱导的IOP升高所必需的。 表达可能改变房水流出的基因的慢病毒载体已被注射到活猴眼睛的前段。 在一只眼睛中转导C3转移酶（能够改变肌动蛋白细胞骨架）不会改变IOP。 正在检查眼组织以验证存在表达。 另一个基因，前列腺素合成酶（预期增加葡萄膜巩膜流出），被输送到5只猴子的每一只眼睛的前段。 在注射后2周至5个月，在5只眼中的3只中检测到IOP一过性降低。 绿色荧光蛋白的共表达在体内可检测到近2年。将检查组织的PGF合酶表达。 维生素D局部应用于眼睛降低IOP。 目前正在研究这种减少的机制。 这项研究使用WNPRC研究服务。 出版物： Lee E，Gabelt BT，Faralli JA，Peters DM，Brandt CR，考夫曼PL，Bhattacharya SK：培养的人小梁网细胞中腺病毒的COCH转基因表达及其对猴器官培养前段流出功能的影响。 Invest Ophthalmol维斯科学，电子版，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.