Regeneration of mammalian cochlear and vestibular hair cells through Hes1/Hes5 modulation with siRNA

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• 作者：Xiaoping Du ; Wei Li ; Xinsheng Gao ; Matthew B. West ; W. Mark Saltzman ; Christopher J. Cheng ; Charles Stewart ; Jie Zheng ; Weihua Cheng ; Richard D. Kopke
• 关键词：Hair cell regeneration ; Inner ear ; Notch pathway ; siRNA ; Nanoparticle ; Mouse
• 刊名：Hearing Research
• 出版年：2013
• 出版时间：October, 2013
• 年：2013
• 卷：304
• 期：Complete
• 页码：91-110
• 全文大小：8465 K

文摘

The Notch pathway is a cell signaling pathway determining initial specification and subsequent cell fate in the inner ear. Previous studies have suggested that new hair cells (HCs) can be regenerated in the inner ear by manipulating the Notch pathway. In the present study, delivery of siRNA to m>Hes1m> and m>Hes5m> using a transfection reagent or siRNA to m>Hes1m> encapsulated within poly(lactide-m>com>-glycolide acid) (PLGA) nanoparticles increased HC numbers in non-toxin treated organotypic cultures of cochleae and maculae of postnatal day 3 mouse pups. An increase in HCs was also observed in cultured cochleae and maculae of mouse pups pre-conditioned with a HC toxin (4-hydroxy-2-nonenal or neomycin) and then treated with the various siRNA formulations. Treating cochleae with siRNA to m>Hes1m> associated with a transfection reagent or siRNA to m>Hes1m> delivered by PLGA nanoparticles decreased m>Hes1m> mRNA and up-regulated m>Atoh1m> mRNA expression allowing supporting cells (SCs) to acquire a HC fate. Experiments using cochleae and maculae of p27^kip1/-GFP transgenic mouse pups demonstrated that newly generated HCs trans-differentiated from SCs. Furthermore, PLGA nanoparticles are non-toxic to inner ear tissue, readily taken up by cells within the tissue of interest, and present a synthetic delivery system that is a safe alternative to viral vectors. These results indicate that when delivered using a suitable vehicle, m>Hesm> siRNAs are potential therapeutic molecules that may have the capacity to regenerate new HCs in the inner ear and possibly restore human hearing and balance function.