Institute of Clinical Medicine, National Yang-Ming University, Taipei.
Department of Ophthalmology, Taipei Veterans General Hospital, Taipei.
Cell Transplant. 2019 Nov;28(11):1345-1357. doi: 10.1177/0963689719860130. Epub 2019 Jul 17.
Best dystrophy (BD), also termed best vitelliform macular dystrophy (BVMD), is a juvenile-onset form of macular degeneration and can cause central visual loss. Unfortunately, there is no clear definite therapy for BD or improving the visual function on this progressive disease. The human induced pluripotent stem cell (iPSC) system has been recently applied as an effective tool for genetic consultation and chemical drug screening. In this study, we developed patient-specific induced pluripotent stem cells (BD-iPSCs) from BD patient-derived dental pulp stromal cells and then differentiated BD-iPSCs into retinal pigment epithelial cells (BD-RPEs). BD-RPEs were used as an expandable platform for in vitro candidate drug screening. Compared with unaffected sibling-derived iPSC-derived RPE cells (Ctrl-RPEs), BD-RPEs exhibited typical RPE-specific markers with a lower expression of the tight junction protein ZO-1 and Bestrophin-1 (BEST1), as well as reduced phagocytic capabilities. Notably, among all candidate drugs, curcumin was the most effective for upregulating both the BEST1 and ZO-1 genes in BD-RPEs. Using the iPSC-based drug-screening platform, we further found that curcumin can significantly improve the mRNA expression levels of Best gene in BD-iPSC-derived RPEs. Importantly, we demonstrated that curcumin-loaded PLGA nanoparticles (Cur-NPs) were efficiently internalized by BD-RPEs. The Cur-NPs-based controlled release formulation further increased the expression of ZO-1 and Bestrophin-1, and promoted the function of phagocytosis and voltage-dependent calcium channels in BD-iPSC-derived RPEs. We further demonstrated that Cur-NPs enhanced the expression of antioxidant enzymes with a decrease in intracellular ROS production and hydrogen peroxide-induced oxidative stress. Collectively, these data supported that Cur-NPs provide a potential cytoprotective effect by regulating the anti-oxidative abilities of degenerated RPEs. In addition, the application of patient-specific iPSCs provides an effective platform for drug screening and personalized medicine in incurable diseases.
最佳营养不良症（BD），也称为最佳类玻璃体黄斑营养不良症（BVMD），是一种青少年发病的黄斑变性形式，可导致中心视力丧失。不幸的是，对于 BD 或改善这种进行性疾病的视力功能，目前尚无明确的治疗方法。人类诱导多能干细胞（iPSC）系统最近已被应用于遗传咨询和化学药物筛选的有效工具。在这项研究中，我们从 BD 患者来源的牙髓基质细胞中开发了患者特异性诱导多能干细胞（BD-iPSC），然后将 BD-iPSC 分化为视网膜色素上皮细胞（BD-RPE）。BD-RPE 被用作体外候选药物筛选的可扩展平台。与未受影响的兄弟姐妹来源的 iPSC 衍生的 RPE 细胞（Ctrl-RPE）相比，BD-RPE 表现出典型的 RPE 特异性标记，其紧密连接蛋白 ZO-1 和 Bestrophin-1（BEST1）的表达降低，吞噬能力降低。值得注意的是，在所有候选药物中，姜黄素对 BD-RPE 中 BEST1 和 ZO-1 基因的上调最有效。使用基于 iPSC 的药物筛选平台，我们进一步发现姜黄素可以显著提高 BD-iPSC 衍生的 RPE 中 Best 基因的 mRNA 表达水平。重要的是，我们证明姜黄素负载的 PLGA 纳米颗粒（Cur-NPs）可被 BD-RPE 有效内化。Cur-NPs 为基础的控释制剂进一步增加了 ZO-1 和 Bestrophin-1 的表达，并促进了 BD-iPSC 衍生的 RPE 中的吞噬作用和电压依赖性钙通道的功能。我们进一步证明 Cur-NPs 通过调节变性 RPE 的抗氧化能力来增强抗氧化酶的表达，同时减少细胞内 ROS 产生和过氧化氢诱导的氧化应激。总的来说，这些数据表明 Cur-NPs 通过调节变性 RPE 的抗氧化能力提供了潜在的细胞保护作用。此外，患者特异性 iPSC 的应用为不可治愈疾病的药物筛选和个性化医疗提供了有效的平台。
