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.

该副本是使用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这是调查员的机构。 目的：确定产生一氧化氮（NO）的化合物是否可以在体外放松卡尔巴乔尔收缩或静止的睫状肌（CM）。这表明该类别的化合物是否可能有助于增强葡萄球菌流出作为降低青光眼中眼内压（IOP）的方法。确定化合物对体外CM合同/放松的影响以及体内IOP。利用器官培养中的猴子前部段来研究基因疗法和其他分子对小梁输出流的影响，这对于青光眼治疗也可能很重要。确定体内基因疗法对前部段的影响。改变水性幽默输出流的蛋白质过表达可能导致眼内减压，这对于青光眼治疗很重要。或者，眼内压的升高可能导致新的青光眼模型。 进展：在给药一氧化氮供体之前，通过用ODQ（ODQ）预处理ODQ（ODQ）预处理CM松弛响应。在需要CM松弛或收缩的治疗区域，例如在青光眼治疗中，任何化合物都没有潜在的价值。 猴子器官培养的前段（MOCA）系统被用于确定药物治疗和基因治疗对小梁输出流的影响，该系统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的过表达会增加眼内压（IOP）并减少小梁输出流。通过在培养基中纳入牛血清白蛋白，可以增强使用转化生长Beta-2（TGF¿2）治疗后MOCAS中的IOP升高反应。在TGF¿2处理过程中，沉默的Cochlin表达的研究将确定Cochlin诱导是否需要TGF诱导的IOP升高。 表达可能改变水性幽默出口的基因的慢病毒载体已注入活着的猴子眼前部分。一只眼睛中C3转移酶（能够改变肌动蛋白细胞骨架）的转导不会改变IOP。正在检查眼组织以验证表达。另一种基因，前列腺素合酶（预计将增强葡萄球菌出口），被传递到5只猴子中每只眼睛的一只眼睛的前部。在注射后2周到5个月，在5眼中3眼中3眼中3眼中3眼中检测到IOP的短暂降低。在体内检测到绿色荧光蛋白的共表达近2年。将检查组织的PGFSENTHase表达。 维生素D局部应用于眼睛降低IOP。正在研究这种减少的机制。 这项研究使用了WNPRC研究服务。 发布： Lee E，Gabelt BT，Faralli JA，Peters DM，Brandt CR，Kaufman PL，Bhattacharya SK：腺病毒在培养的人类小梁网细胞中腺病毒的COCH转化表达及其对猴子器官培养前段的产量设施的影响。 2009年11月20日，Invest Ophthalmol Vis Sci，Epub发行。