Yang Yi-Ping, Chang Yuh-Lih, Lai Yun-Hsien, Tsai Ping-Hsing, Hsiao Yu-Jer, Nguyen Long Hoang, Lim Xue-Zhen, Weng Chang-Chi, Ko Yu-Ling, Yang Chang-Hao, Hwang De-Kuang, Chen Shih-Jen, Chiou Shih-Hwa, Chiou Guang-Yuh, Wang An-Guor, Chien Yueh
Department of Medical Research, Taipei Veteran General Hospital, Taipei 11217, Taiwan.
Institute of Food Safety and Health Risk Assessment, School of Pharmaceutical Sciences, National Yang Ming Chiao Tung University, Taipei 11217, Taiwan.
Biomedicines. 2022 Mar 28;10(4):788. doi: 10.3390/biomedicines10040788.
: Leber's hereditary optic neuropathy (LHON) is known as an inherited retinal disorder characterized by the bilateral central vision loss and degeneration of retinal ganglion cells (RGCs). Unaffected LHON carriers are generally asymptomatic, suggesting that certain factors may contribute to the disease manifestations between carriers and patients who carry the same mutated genotypes. Methods: We first aimed to establish the iPSC-differentiated RGCs from the normal healthy subject, the carrier, and the LHON patient and then compared the differential expression profile of circular RNAs (CircRNAs) among RGCs from these donors in vitro. We further overexpressed or knocked down the most upregulated circRNA to examine whether this circRNA contributes to the distinct phenotypic manifestations between the carrier- and patient-derived RGCs. : iPSCs were generated from the peripheral blood cells from the healthy subject, the carrier, and the LHON patient and successfully differentiated into RGCs. These RGCs carried equivalent intracellular reactive oxygen species, but only LHON-patient iPSC-derived RGCs exhibited remarkable apoptosis. Next-generation sequencing and quantitative real-time PCR revealed the circRNA hsa_circ_0087207 as the most upregulated circRNA in LHON-patient iPSC-derived RGCs. Overexpression of hsa_circ_0087207 increased the apoptosis in carrier iPSC-derived RGCs, while knockdown of hsa_circ_0087207 attenuated the apoptosis in LHON-patient iPSC-derived RGCs. Predicted by bioinformatics approaches, hsa_circ_0087207 acts as the sponge of miR-665 to induce the expression of a variety of apoptosis-related genes in LHON patient iPSC-derived RGCs. : Our data indicated that hsa_circ_0087207 upregulation distinguishes the disease phenotype manifestations between iPSC-derived RGCs generated from the LHON patient and carrier. Targeting the hsa_circ_0087207/miR-665 axis might hold therapeutic promises for the treatment of LHON.
莱伯遗传性视神经病变（LHON）是一种遗传性视网膜疾病，其特征为双侧中心视力丧失以及视网膜神经节细胞（RGCs）变性。未受影响的LHON携带者通常无症状，这表明某些因素可能导致携带相同突变基因型的携带者和患者之间出现疾病表现差异。方法：我们首先旨在从正常健康受试者、携带者和LHON患者中建立诱导多能干细胞（iPSC）分化的RGCs，然后在体外比较这些供体来源的RGCs中环状RNA（CircRNAs）的差异表达谱。我们进一步过表达或敲低上调最明显的环状RNA，以检查该环状RNA是否导致携带者和患者来源的RGCs之间出现明显的表型差异。从健康受试者、携带者和LHON患者的外周血细胞中生成了iPSCs，并成功分化为RGCs。这些RGCs具有等量的细胞内活性氧，但只有LHON患者iPSC来源的RGCs表现出明显的凋亡。下一代测序和定量实时PCR显示，环状RNA hsa_circ_0087207是LHON患者iPSC来源的RGCs中上调最明显的环状RNA。过表达hsa_circ_0087207会增加携带者iPSC来源的RGCs中的凋亡，而敲低hsa_circ_0087207则会减弱LHON患者iPSC来源的RGCs中的凋亡。通过生物信息学方法预测，hsa_circ_0087207作为miR-665的海绵，可诱导LHON患者iPSC来源的RGCs中多种凋亡相关基因的表达。我们的数据表明，hsa_circ_0087207的上调区分了LHON患者和携带者产生的iPSC来源的RGCs之间的疾病表型表现。靶向hsa_circ_0087207/miR-665轴可能为LHON的治疗带来治疗前景。
