title: A Small Indel Mutant Mouse Model of Epidermolytic Palmoplantar Keratoderma and Its Application to Mutant-specific shRNA Therapy.
abstract: Epidermolytic palmoplantar keratoderma (EPPK) is a relatively common autosomal-dominant skin disorder caused by mutations in the keratin 9 gene (KRT9), with few therapeutic options for the affected so far. Here, we report a knock-in transgenic mouse model that carried a small insertion-deletion (indel) mutant of Krt9, c.434delAinsGGCT (p.Tyr144delinsTrpLeu), corresponding to the human mutation KRT9/c.500delAinsGGCT (p.Tyr167delinsTrpLeu), which resulted in a human EPPK-like phenotype in the weight-stress areas of the fore- and hind-paws of both Krt9(+/mut) and Krt9(mut/mut) mice. The phenotype confirmed that EPPK is a dominant-negative condition, such that mice heterozygotic for the K9-mutant allele (Krt9(+/mut)) showed a clear EPPK-like phenotype. Then, we developed a mutant-specific short hairpin RNA (shRNA) therapy for EPPK mice. Mutant-specific shRNAs were systematically identified in vitro using a luciferase reporter gene assay and delivered into Krt9(+/mut) mice. shRNA-mediated knockdown of mutant protein resulted in almost normal morphology and functions of the skin, whereas the same shRNA had a negligible effect in wild-type K9 mice. Our results suggest that EPPK can be treated by gene therapy, and this has significant implications for future clinical application.
authors.affiliation: Department of Cell Biology and Medical Genetics, Research Center for Molecular Medicine, National Education Base for Basic Medical Sciences, Institute of Cell Biology, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Ya-Su
authors.initials: YS
authors.lastname: Lyu
authors.affiliation: Key Laboratory of Model Animals for Disease Study of The Ministry of Education, Model Animal Research Center of Nanjing University, Nanjing, Jiangsu, China.
authors.forename: Pei-Liang
authors.initials: PL
authors.lastname: Shi
authors.affiliation: Department of Biological Chemistry, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, China.
authors.forename: Xiao-Ling
authors.initials: XL
authors.lastname: Chen
authors.affiliation: Department of Cell Biology and Medical Genetics, Research Center for Molecular Medicine, National Education Base for Basic Medical Sciences, Institute of Cell Biology, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Yue-Xiao
authors.initials: YX
authors.lastname: Tang
authors.affiliation: Department of Cell Biology and Medical Genetics, Research Center for Molecular Medicine, National Education Base for Basic Medical Sciences, Institute of Cell Biology, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Yan-Fang
authors.initials: YF
authors.lastname: Wang
authors.affiliation: Department of Cell Biology and Medical Genetics, Research Center for Molecular Medicine, National Education Base for Basic Medical Sciences, Institute of Cell Biology, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Rong-Rong
authors.initials: RR
authors.lastname: Liu
authors.affiliation: Department of Cell Biology and Medical Genetics, Research Center for Molecular Medicine, National Education Base for Basic Medical Sciences, Institute of Cell Biology, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Xiao-Rui
authors.initials: XR
authors.lastname: Luan
authors.affiliation: Experimental Teaching Centre for Basic Medical Sciences, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Fang
authors.affiliation: Experimental Teaching Centre for Basic Medical Sciences, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Ru-Huan
authors.initials: RH
authors.lastname: Mei
authors.affiliation: Department of Cell Biology and Medical Genetics, Research Center for Molecular Medicine, National Education Base for Basic Medical Sciences, Institute of Cell Biology, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Zhen-Fang
authors.initials: ZF
authors.lastname: Du
authors.affiliation: Department of Biology, Ningbo College of Health Sciences, Ningbo, Zhejiang, China.
authors.forename: Hai-Ping
authors.initials: HP
authors.lastname: Ke
authors.affiliation: Department of Medicine, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Erik
authors.initials: E
authors.lastname: Matro
authors.affiliation: Key Laboratory of Model Animals for Disease Study of The Ministry of Education, Model Animal Research Center of Nanjing University, Nanjing, Jiangsu, China.
authors.forename: Ling-En
authors.initials: LE
authors.lastname: Li
authors.affiliation: Key Laboratory of Model Animals for Disease Study of The Ministry of Education, Model Animal Research Center of Nanjing University, Nanjing, Jiangsu, China.
authors.forename: Zhao-Yu
authors.initials: ZY
authors.lastname: Lin
authors.affiliation: Key Laboratory of Model Animals for Disease Study of The Ministry of Education, Model Animal Research Center of Nanjing University, Nanjing, Jiangsu, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Model Animals for Disease Study of The Ministry of Education, Model Animal Research Center of Nanjing University, Nanjing, Jiangsu, China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Gao
authors.affiliation: Department of Cell Biology and Medical Genetics, Research Center for Molecular Medicine, National Education Base for Basic Medical Sciences, Institute of Cell Biology, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
authors.forename: Xian-Ning
authors.initials: XN
authors.lastname: Zhang