GLAUCOMA THERAPY, CILIARY MUSCLE CONTRACTION AND TRABECULAR OUTFLOW

青光眼治疗、睫状肌收缩和小梁流出

• 批准号: 8173066
• 负责人: PAUL L KAUFMAN
• 金额: $ 3.1万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8173066
• 关键词: Actins; Adenoviruses; Anterior; Aqueous Humor; Area; Bovine Serum Albumin; Carbachol; Cells; Ciliary Muscle; Computer Retrieval of Information on Scientific Projects Database; Contracts; Cytoskeleton; Eye; Funding; Genes; Glaucoma; Grant; Green Fluorescent Proteins; Growth; Guanylate Cyclase; Human; In Vitro; Injection of therapeutic agent; Institution; Lead; Lentivirus Vector; Life; Modeling; Monkeys; Muscle Contraction; Muscle relaxation phase; Nitric Oxide; Nitric Oxide Donors; Organ Culture Techniques; Pathway interactions; Pharmacotherapy; Physiologic Intraocular Pressure; Printing; Prostaglandin-Endoperoxide Synthase; Protein Overexpression; Publications; Relaxation; Research; Research Personnel; Resources; Rest; Services; Source; System; Therapeutic; Tissues; Topical application; Trabecular meshwork structure; Transferase; United States National Institutes of Health; Vitamin D; gene therapy; in vivo; inhibitor/antagonist; response; transgene expression

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.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以出现在其他 CRISP 条目中 列出的机构是。 对于中心来说，它不一定是研究者的机构。 目的：确定产生一氧化氮（NO）的化合物是否可以在体外松弛收缩的或静止的睫状肌（CM），这是此类化合物是否可用于增强葡萄膜巩膜流出作为降低眼内的方法的指标。青光眼眼压（IOP） 确定化合物对体外 CM 收缩/舒张以及体内 IOP 的影响 利用器官培养中的猴子眼前节来研究基因的影响。治疗和其他分子对小梁流出物的影响，这对青光眼治疗也很重要确定体内基因治疗对眼前节的影响，改变房水流出的蛋白质可能导致眼压降低，这对青光眼治疗很重要。或者，眼内压升高可能导致新的青光眼模型。 进展：在给予一氧化氮供体之前，用鸟苷酸环化酶途径抑制剂 ODQ 进行预处理，可阻断 CM 对一氧化氮供体的松弛反应，该化合物在需要 CM 松弛或收缩的治疗领域具有潜在价值。 ，例如青光眼的治疗。 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，耳蜗蛋白的过度表达仅在眼内升高。人青光眼，增加眼内压 (IOP) 并减少小梁流出。通过添加转化生长β-2 (TGF¿2) 治疗后，MOCAS 中的 IOP 升高反应得到增强。培养基中牛血清白蛋白抑制 TGF 期间 cochlin 表达的研究。 2 治疗将确定 TGF 是否需要 cochlin 诱导？ 2引起眼压升高。 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体猴子眼前节中，目前正在检查在一只眼睛中转导 C3 转移酶（能够改变肌动蛋白细胞骨架）不会改变眼组织。验证表达是否存在，前列腺素合酶（预计会增强葡萄膜巩膜流出），被传递到 5 只猴子 A 中每只的一只眼睛的眼前节。注射后 2 周至 5 个月，5 只眼睛中的 3 只检测到 IOP 短暂降低。在体内可检测到绿色荧光蛋白的共表达，并将检查组织的 PGF 合酶表达。 局部使用维生素 D 可降低眼压，目前正在研究这种降低的机制。 本研究使用了 WNPRC 研究服务。 发布： Lee E、Gabelt BT、Faralli JA、Peters DM、Brandt CR、Kaufman PL、Bhattacharya SK：腺病毒在培养的人小梁网细胞中的 COCH 转基因表达及其对猴子器官培养的前段流出设施的影响 投资眼科视觉科学。 ，印刷前的电子版，2009 年 11 月 20 日。