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.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 目的：确定维生素D对眼内压（IOP）的影响 猴子在体内。 利用器官培养（MOCA）中的猴子前部 基因疗法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 幽默流出可能导致眼内降低，这对于 青光眼治疗。 或者，眼内高海拔可能导致新的 青光眼模型。 描述： 在体内用维生素D局部处理猴子的眼睛，对IOP的影响 确定。 蛋白质Cochlin的过表达，仅在人类青光眼中升高， 增加眼内压（IOP）并减少小梁流出。 用TGFB2处理MOCA会诱导Cochlin产生。 科克林在 TGFB2诱导的IOP升高将通过沉默的Cochlin表达来确定 在TGFB2处理过程中，用SHRNA用于Cochlin。 表达Caldesmon或C3的Adeno病毒载体已显示可减少IOP mocas。 在MOCAS中测试了表达Caldesmon或C3的F I V vector 在体内注射到猴子眼中。 用纤连蛋白的HEPII结构域处理MOCAS，以评估 IOP还原过程中小梁网的形态。 进步： 维生素D的体内猴子眼睛的局部治疗降低了IOP。 这 这种作用的机制正在研究中。 似乎没有 对葡萄球菌流出或水性幽默形成的影响。 用mocas处理 VITD对IOP没有任何影响，表明小梁流出不受影响。 用表达shRNA Cochlin或负的H I V量处理MOCAS 控制。 没有影响的眼内压或流出设施 仅向量。 TGFB2在MOCAS中诱导的IOP升高不够一致 在配对的MOCA之间确定沉默Cochlin表达对 TGFB2诱导IOP升高。 将使用Adeno进行其他实验 载体过表达的Cochlin。 用表达Caldesmon或C3的F I V向量对MOCA进行处理不会降低IOP。 先前使用Adeno病毒载体结果的可能性部分可能是由于 通过与Adeno矢量相互作用的Caldesmon或C3响应的增强 整联蛋白正在研究。 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 囊泡从其靶标隔离C3。 HEPII治疗MOCA导致内壁之间的空间扩展 与对照相比，Schlemm的运河和小梁胶原蛋白束相比。 这项研究使用了WNPRC动物和病理服务。 发布： *Schwinn MK，Gonzalez JM JR，Gabelt BT，Sheibani N，Kaufman PL，Peters DM：肝素 纤连蛋白的II结构域通过α4Beta1co-Signaling介导收缩性 路径。 Exp Eye Res，316：1500-1512，2010年。PMID：20302860，PMCID2871963。