title: Regeneration of mammalian cochlear and vestibular hair cells through Hes1/Hes5 modulation with siRNA.
abstract: 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 Hes1 and Hes5 using a transfection reagent or siRNA to Hes1 encapsulated within poly(lactide-co-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 Hes1 associated with a transfection reagent or siRNA to Hes1 delivered by PLGA nanoparticles decreased Hes1 mRNA and up-regulated Atoh1 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, Hes 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.
authors.affiliation: Hough Ear Institute, P.O. Box 23206, Oklahoma City, OK 73112, USA.
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Du
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.forename: Xinsheng
authors.initials: X
authors.lastname: Gao
authors.forename: Matthew B
authors.initials: MB
authors.lastname: West
authors.forename: W Mark
authors.initials: WM
authors.lastname: Saltzman
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Cheng
authors.forename: Charles
authors.initials: C
authors.lastname: Stewart
authors.forename: Jie
authors.initials: J
authors.lastname: Zheng
authors.forename: Weihua
authors.initials: W
authors.lastname: Cheng
authors.forename: Richard D
authors.initials: RD
authors.lastname: Kopke
mesh_terms: D000447:Aldehydes / Q000633:toxicity; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics*; D006198:Hair Cells, Auditory / Q000187:drug effects / Q000502:physiology*; D018069:Hair Cells, Vestibular / Q000187:drug effects / Q000502:physiology*; D018398:Homeodomain Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics*; D006801:Humans; D019344:Lactic Acid; D051379:Mice; D008822:Mice, Transgenic; D053758:Nanoparticles; D009355:Neomycin / Q000633:toxicity; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics; D051880:Receptors, Notch / Q000502:physiology; D012038:Regeneration / Q000235:genetics* / Q000502:physiology*; D012097:Repressor Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics*; D046509:Tissue Culture Techniques; D000072056:Transcription Factor HES-1
chemical_list: D000447:Aldehydes; D051792:Basic Helix-Loop-Helix Proteins; D018398:Homeodomain Proteins; D019344:Lactic Acid; D009355:Neomycin; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051880:Receptors, Notch; D012097:Repressor Proteins; D000072056:Transcription Factor HES-1; C500943:Atoh1 protein, mouse; C489734:Hes1 protein, mouse; C489685:Hes5 protein, mouse; C027576:4-hydroxy-2-nonenal
keywords: 4-HNE; 4-hydroxy-2-nonenal; GFP; HC; Hair cell regeneration; Hes1 and Hes5; Inner ear; LF; Mouse; NP; Nanoparticle; Notch pathway; OC; PLGA; SC; green fluorescent protein; hair cell; hairy and enhancer of split 1 and 5; lipofection; nanoparticle; poly(lactide-co-glycolide acid); qRT-PCR; quantitative real-time polymerase chain reaction; scRNA; scrambled siRNA; siRNA; small interfering RNA; supporting cell; the organ of Corti