GLAUCOMA THERAPY, CILIARY MUSCLE CONTRACTION AND TRABECULAR OUTFLOW

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 目的：探讨维生素D对老年人眼压的影响。 活体中的猴子。 利用猕猴前段器官培养技术(MOCAS)研究其在器官培养中的作用 基因治疗和其他分子对小梁流出的影响 对青光眼治疗很重要。改变房水的蛋白质的过度表达 体液流出可能导致眼压下降，这对 青光眼治疗。或者，眼压升高可能会导致新的 青光眼模型。 说明： 在体猴眼局部应用维生素D及其对眼压的影响 已经下定决心。 Cochlin蛋白的过度表达，这种蛋白只在人类青光眼中升高， 增加眼压(IOP)，减少小梁流出。 用TGFb2处理MOCAS可诱导椰子碱的产生。Cochlin在人类进化过程中的作用 TGFb2诱导的眼压升高将通过沉默Cochlin的表达来确定 在TGFb2治疗过程中用shRNA表达cochlin。 表达钙调蛋白或补体C3的腺病毒载体被证明能降低眼压 莫卡斯。表达caldesmon或C3的F I V载体之前在MOCAS中进行了测试 注射入猴眼体内。 用纤维连接蛋白的HepII结构域治疗MOCAS，以评价 降眼压过程中小梁网的形态变化。 进度： 体内用维生素D局部治疗猴眼可降低眼压。这个 这种效应的机制正在调查中。似乎没有任何 对葡萄膜巩膜流出或房水形成的影响。多器官功能不全综合征的治疗 维生素D对眼压无影响，提示小梁流出不受影响。 用表达shRNA cochlin或阴性的H I V载体处理MOCAS 控制力。对眼内压或流出设施没有影响。 仅载体一项。TGFb2诱导的MOCAS眼压升高不够一致 在配对的MOCAS之间确定沉默cochlin表达的影响 TGFb2诱导眼压升高。将对腺病毒进行更多的实验 载体过度表达Cochlin。 用表达钙调蛋白或补体C3的F I V载体治疗MOCAS不能降低眼压。 以前对腺病毒载体的结果可能部分归因于 腺病毒载体相互作用增强钙调蛋白或补体C3应答 整合素正在接受研究。细胞可溶性C3也不能降低MOCAS的眼压。 商业上可获得C3上的TAT序列可能导致内吞作用进入 囊泡将C3与其目标隔离。 HEII治疗MOCAS导致内壁间隙扩大 将Schlemm管和小梁胶原梁与对照组进行比较。 这项研究使用了WNPRC动物和病理服务中心。 出版物： *Schwinn MK，Gonzalez JM Jr，Gabelt BT，Sheibani N，Kaufman PL，彼得斯DM：肝素 纤维连接蛋白II结构域通过α4β1共信号调节收缩 路径。EXP Eye Res，316：1500-1512,2010。PMID：20302860，PMCID2871963。

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.