Cho In Hwan, Park Seong Jun, Lee Si Hyung, Nah Seung Kwan, Park Ha Yan, Yang Jin Young, Madrakhimov Sanjar Batirovich, Lyu Jungmook, Park Tae Kwann
Department of Ophthalmology, Soonchunhyang University Hospital Cheonan, Cheonan, Republic of Korea.
College of Medicine, Soonchunhyang University, Cheonan, Republic of Korea.
Lasers Med Sci. 2019 Apr;34(3):571-581. doi: 10.1007/s10103-018-2631-5. Epub 2018 Sep 22.
To investigate the role of Wnt/β-catenin signaling pathway in the restoration of induced pluripotent stem cell-derived retinal pigment epithelium (hiPSC-RPE) after laser photocoagulation. After differentiation of RPE cells from hiPSCs, laser photocoagulation was performed. Activation of Wnt/β-catenin signaling at days 1 and 5 after laser photocoagulation was evaluated by expression of β-catenin. Cell proliferation and alteration in cell-to-cell contact at day 5 after laser photocoagulation with or without Dickkopf-1 (Dkk-1) treatment were studied using ethynyl-2'-deoxyuridine (EdU) assay and zonula occludens-1 (ZO-1) expression analysis, respectively. The mRNA levels of Wnt genes at day 5 after laser photocoagulation were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). Activation of Wnt/β-catenin signaling at days 1 and 5 after laser photocoagulation was confirmed by β-catenin accumulation in the cytoplasm and nucleus of hiPSC-RPE. Many EdU-positive cells also expressed β-catenin, and the number of EdU-positive cells was decreased at day 5 after laser photocoagulation after Dkk-1 treatment, indicating that Wnt/β-catenin signaling mediated hiPSC-RPE proliferation. ZO-1 expression was not decreased with Dkk-1 treatment at day 5 after laser photocoagulation, indicating that Wnt/β-catenin signaling mediated hiPSC-RPE restoration. At day 5, after laser photocoagulation, mRNA levels of Wnt2b, Wnt3, Wnt5a, Wnt7a, and Wnt10b were increased. Wnt/β-catenin signaling has a crucial role in restoration of hiPSC-RPE proliferation after laser photocoagulation. Manipulation of Wnt/β-catenin signaling while elucidating the underlying mechanisms of RPE restoration might have a therapeutic potential in retinal degenerative diseases.
研究Wnt/β-连环蛋白信号通路在激光光凝后诱导多能干细胞来源的视网膜色素上皮细胞（hiPSC-RPE）修复中的作用。从hiPSC分化出RPE细胞后，进行激光光凝。通过β-连环蛋白的表达评估激光光凝后第1天和第5天Wnt/β-连环蛋白信号的激活情况。分别使用乙炔基-2'-脱氧尿苷（EdU）检测和紧密连接蛋白-1（ZO-1）表达分析，研究激光光凝后第5天给予或不给予Dickkopf-1（Dkk-1）处理时的细胞增殖和细胞间接触的改变。通过定量实时聚合酶链反应（qRT-PCR）评估激光光凝后第5天Wnt基因的mRNA水平。激光光凝后第1天和第5天Wnt/β-连环蛋白信号的激活通过hiPSC-RPE细胞质和细胞核中β-连环蛋白的积累得以证实。许多EdU阳性细胞也表达β-连环蛋白，并且在Dkk-1处理后激光光凝第5天EdU阳性细胞数量减少，表明Wnt/β-连环蛋白信号介导hiPSC-RPE增殖。激光光凝后第5天给予Dkk-1处理时ZO-1表达未降低，表明Wnt/β-连环蛋白信号介导hiPSC-RPE修复。在激光光凝后第5天，Wnt2b、Wnt3、Wnt5a、Wnt7a和Wnt10b的mRNA水平升高。Wnt/β-连环蛋白信号在激光光凝后hiPSC-RPE增殖的修复中起关键作用。在阐明RPE修复的潜在机制的同时操纵Wnt/β-连环蛋白信号可能对视网膜退行性疾病具有治疗潜力。
