Therapeutic potential of AAV-mediated MMP-3 secretion from corneal endothelium in treating glaucoma.
Therapeutic potential of AAV-mediated MMP-3 secretion from corneal endothelium in treating glaucoma.
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DOI：
10.1093/hmg/ddx028
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发表时间：
2017-04-01
影响因子：
3.5
通讯作者：
Humphries P
中科院分区：
文献类型：
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作者：
O'Callaghan J;Crosbie DE;Cassidy PS;Sherwood JM;Flügel-Koch C;Lütjen-Drecoll E;Humphries MM;Reina-Torres E;Wallace D;Kiang AS;Campbell M;Stamer WD;Overby DR;O'Brien C;Tam LCS;Humphries P
眼内压(IOP)的维持是由于睫状突产生的房水(AH)与其流出的流体动力学阻力之间的平衡，流出的流出途径包括小梁网(TM)和Schlemm管(SC)。高眼压是开角型青光眼的主要危险因素，高眼压可由高眼压引起。基质金属蛋白酶(MMPs)对传统的房水流出动态平衡有重要作用，它可以重塑细胞外基质，直接影响房水流出阻力和眼压。我们观察到，与年龄匹配的正常血压对照组相比，青光眼急性发作组的基质金属蛋白酶-3活性降低。与对照组或外源性基质金属蛋白酶-3治疗组相比，青光眼治疗组的SC内皮细胞和TM细胞单层的跨内皮阻力显著增加，单层通透性降低。野生型小鼠体内接种含有CMV驱动的基质金属蛋白酶-3基因的AAV-2/9(AAV-MMP3)后，可有效地转导角膜内皮细胞，并增加基质金属蛋白酶-3的水浓度和活性。最重要的是，AAV介导的基质金属蛋白酶-3的表达增加了流出的便利性，降低了眼压，使用局部注射多西环素激活的可诱导启动子控制表达也达到了同样的效果。用透射电子显微镜对基质基质进行超微结构分析，发现核心细胞外基质成分发生重塑和降解。这些结果表明，通过使用眼药水，周期性地诱导AAV介导的MMP-3分泌到AH中，对于那些对传统降压药物疗效不佳的青光眼病例可能具有治疗潜力。
Intraocular pressure (IOP) is maintained as a result of the balance between production of aqueous humour (AH) by the ciliary processes and hydrodynamic resistance to its outflow through the conventional outflow pathway comprising the trabecular meshwork (TM) and Schlemm’s canal (SC). Elevated IOP, which can be caused by increased resistance to AH outflow, is a major risk factor for open-angle glaucoma. Matrix metalloproteinases (MMPs) contribute to conventional aqueous outflow homeostasis in their capacity to remodel extracellular matrices, which has a direct impact on aqueous outflow resistance and IOP. We observed decreased MMP-3 activity in human glaucomatous AH compared to age-matched normotensive control AH. Treatment with glaucomatous AH resulted in significantly increased transendothelial resistance of SC endothelial and TM cell monolayers and reduced monolayer permeability when compared to control AH, or supplemented treatment with exogenous MMP-3. Intracameral inoculation of AAV-2/9 containing a CMV-driven MMP-3 gene (AAV-MMP-3) into wild type mice resulted in efficient transduction of corneal endothelium and an increase in aqueous concentration and activity of MMP-3. Most importantly, AAV-mediated expression of MMP-3 increased outflow facility and decreased IOP, and controlled expression using an inducible promoter activated by topical administration of doxycycline achieved the same effect. Ultrastructural analysis of MMP-3 treated matrices by transmission electron microscopy revealed remodelling and degradation of core extracellular matrix components. These results indicate that periodic induction, via use of an eye drop, of AAV-mediated secretion of MMP-3 into AH could have therapeutic potential for those cases of glaucoma that are sub-optimally responsive to conventional pressure-reducing medications.