Reprogramming of endogenous Müller cells into photoreceptor-like cells induced by small-molecule compounds in mice
Reprogramming of endogenous Müller cells into photoreceptor-like cells induced by small-molecule compounds in mice
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DOI：
10.21203/rs.3.rs-890527/v1
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发表时间：
2021-09
期刊：
影响因子：
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通讯作者：
Yuya Fujii;Mitsuru Arima;Y. Murakami;Koh-hei Sonoda
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作者：
Yuya Fujii;Mitsuru Arima;Y. Murakami;Koh-hei Sonoda
终身视力障碍是由于哺乳动物的感光细胞不能再生而导致的视网膜疾病。我们证明了通过玻璃体内注射四种小分子化合物：肿瘤生长因子-β抑制剂、骨形态发生蛋白抑制剂、糖原合成酶激酶3抑制剂和γ-分泌酶抑制剂，小鼠内源性Müller细胞（MCs）可分化为光感受器样细胞。在体外，在MCs的视紫红质（Rho）的信使RNA增加30倍，25%的培养MCs表达Rho蛋白刺激后7天。在体内，Rho阳性细胞在玻璃体内注射四种化合物后第7天再生，伴随着Rho源性暗视功能的恢复。在感光细胞变性的疾病模型N-甲基-N-亚硝基脲治疗的小鼠中，谱系追踪显示再生的Rho阳性细胞来源于内源性MC。最后，Rho阳性细胞的再生也在rd 10小鼠的视网膜中诱导，rd 10小鼠是具有与人类相似的基因突变的模型。值得注意的是，玻璃体内注射显著减少了rd 10小鼠的视锥细胞死亡。这种治疗可能是视网膜再生医学的一种新策略，其中哺乳动物内源性MC被重新编程为感光细胞，而不依赖于移植或基因转移。
Lifelong visual impairment occurs from retinal diseases due to the inability of photoreceptor cells to regenerate in mammals. We demonstrated that endogenous Müller cells (MCs) in mice differentiate into photoreceptor-like cells by intravitreal injection of four small-molecule compounds: tumor growth factor-β inhibitor, bone morphogenetic protein inhibitor, glycogen synthase kinase 3 inhibitor, and γ-secretase inhibitor. In vitro, the messenger RNA of rhodopsin (Rho) in MCs increased 30-fold, and 25% of cultured MCs expressed Rho protein 7 days after stimulation with these compounds. In vivo, Rho-positive cells were regenerated on day 7 after the intravitreal injection of four compounds, accompanied with recovery of Rho-derived scotopic function. Lineage tracing in mice treated with N-methyl-N-nitrosourea, a disease model of photoreceptor degeneration, showed that the regenerated Rho-positive cells were originated from endogenous MCs. Finally, the regeneration of Rho-positive cells was also induced in the retina of rd10 mice, a model with similar genetic mutation as humans. Notably, the intravitreal injection significantly reduced cone cell death in rd10 mice. This treatment could be a new strategy in retinal regenerative medicine where mammalian endogenous MCs are reprogrammed into photoreceptor cells independent of transplantation or gene transfer.