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: 8358194
• 负责人: PAUL L KAUFMAN
• 金额: $ 5.32万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8358194
• 关键词: Adenovirus Vector; Affect; Animals; Anterior; Aqueous Humor; Cells; Ciliary Muscle; Collagen; Endocytosis; Eye; Fibronectins; Funding; Glaucoma; Grant; Heparin; Human; Injection of therapeutic agent; Integrins; Investigation; Lead; Mediating; Modeling; Monkeys; Morphology; Muscle Contraction; National Center for Research Resources; Organ Culture Techniques; Pathology; Physiologic Intraocular Pressure; Primates; Principal Investigator; Production; Research; Research Infrastructure; Resources; Role; Services; Signal Pathway; Source; Structure of sinus venosus of sclera; Testing; Trabecular meshwork structure; United States National Institutes of Health; Vesicle; Viral Vector; Vitamin D; Wisconsin; caldesmon; cost; gene therapy; in vivo; protein expression; research study; response; small hairpin RNA; vector

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Objective: To determine the effects of vitamin D on intraocular pressure (IOP) in monkeys in vivo. To utilize the monkey anterior segment in organ culture (MOCAS) to investigate the effects of gene therapy and other molecules on trabecular outflow which may also be important for glaucoma therapy. Over expression 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. DESCRIPTION: Monkey eyes in vivo were treated topically with Vitamin D and the effects on IOP were determined. Over-expression of the protein cochlin, which is elevated only in human glaucoma, increases intraocular pressure (IOP) and decreases trabecular outflow. Treatment of MOCAS with TGFb2 induces cochlin production. The role of cochlin in the IOP elevation induced by TGFb2 will be determined by silencing cochlin expression with shRNA for cochlin during TGFb2 treatment. Adeno viral vectors expressing caldesmon or C3 have been shown to decrease IOP in MOCAS. F I V vectors expressing caldesmon or C3 are being tested in MOCAS before injection into monkey eyes in vivo. MOCAS were treated with the HepII domain of fibronectin in order to evaluate the morphology of the trabecular meshwork during IOP reduction. PROGRESS: Topical treatment of monkey eyes in vivo with vitamin D decreases IOP. The mechanism for this effect is under investigation. There does not seem to be any effect on uveoscleral outflow or aqueous humor formation. Treatment of MOCAS with VitD did not have any effect on IOP suggesting trabecular outflow is not affected. MOCAS were treated with H I V vectors expressing shRNA cochlin or the negative control. There was no effect out intraocular pressure or outflow facility of the vectors alone. TGFb2 induced IOP elevation in MOCAS was not consistent enough between paired MOCAS to determine the effects of silencing cochlin expression on TGFb2 induced IOP elevation. Additional experiments will be conducted with adeno vectors over-expressing cochlin. Treatment of MOCAS with F I V vectors expressing caldesmon or C3 did not lower IOP. The possibility that previous results with adeno viral vectors may, in part, be due to augmentation of the caldesmon or C3 response by adeno vector interaction with integrins is being investigated. Cell soluble C3 also did not decrease IOP in MOCAS. The TAT sequence on commercially available C3 may have caused endocytosis into vesicles sequestering the C3 from its target. HepII treatment of MOCAS resulted in expansion of the space between the inner wall of Schlemm's canal and the trabecular collagen beams compared to control. This research used WNPRC Animal and Pathology Services. PUBLICATION: *Schwinn MK, Gonzalez JM Jr, Gabelt BT, Sheibani N, Kaufman PL, Peters DM: Heparin II domain of fibronectin mediates contractility through an alpha4beta1co-signaling pathway. Exp Eye Res, 316:1500-1512, 2010. PMID: 20302860, PMCID2871963.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 目的：确定维生素 D 对眼内压 (IOP) 的影响 猴子体内。 利用猴眼前段器官培养（MOCAS）研究 基因治疗和其他分子对小梁流出的影响，这也可能是 对于青光眼治疗很重要。过度表达改变水性的蛋白质 体液流出可能导致眼压降低，这对于 青光眼治疗。 或者，眼压升高可能会导致新的 青光眼模型。 描述： 体内猴眼局部用维生素 D 治疗及其对眼压的影响 被确定。 耳蜗蛋白的过度表达仅在人类青光眼中升高， 增加眼压（IOP）并减少小梁流出。 用 TGFb2 处理 MOCAS 会诱导耳蜗蛋白的产生。 耳蜗素的作用 TGFb2 诱导的 IOP 升高将通过沉默 cochlin 表达来确定 在 TGFb2 治疗期间使用 cochlin 的 shRNA。 表达 caldesmon 或 C3 的腺病毒载体已被证明可以降低眼压 莫卡斯。 表达 caldesmon 或 C3 的 F I V 载体之前正在 MOCAS 中进行测试 注射入猴眼体内。 MOCAS 用纤连蛋白的 HepII 结构域进行处理，以评估 眼压降低过程中小梁网的形态。 进步： 用维生素 D 局部治疗猴眼可降低眼压。 这 这种效应的机制正在研究中。 好像没有 对葡萄膜巩膜流出或房水形成的影响。 MOCAS 的治疗 VitD 对眼压没有任何影响，表明小梁流出不受影响。 MOCAS 用表达 shRNA cochlin 或阴性的 HIV 载体进行处理 控制。 眼压或眼部流出设施没有影响 单独的向量。 MOCAS 中 TGFb2 诱导的 IOP 升高不够一致 配对 MOCAS 之间以确定沉默 cochlin 表达对 TGFb2 诱导 IOP 升高。 将使用腺进行额外的实验 过度表达 cochlin 的载体。 用表达caldesmon或C3的FIV载体处理MOCAS并没有降低IOP。 先前的腺病毒载体结果可能部分归因于 通过腺载体相互作用增强 caldesmon 或 C3 反应 整合素正在研究中。 细胞可溶性 C3 也不会降低 MOCAS 中的 IOP。 市售 C3 上的 TAT 序列可能导致内吞作用 囊泡将 C3 与其靶点隔离。 MOCAS 的 HepII 处理导致内壁之间的空间扩大 与对照相比，施累姆氏管和小梁胶原束的变化。 这项研究使用了 WNPRC 动物和病理学服务。 发布： *Schwinn MK、Gonzalez JM Jr、Gabelt BT、Sheibani N、Kaufman PL、Peters DM：肝素 纤连蛋白 II 结构域通过 alpha4beta1 协同信号介导收缩性 途径。实验眼研究，316：1500-1512，2010。PMID：20302860，PMCID2871963。