title: Human COL7A1-corrected induced pluripotent stem cells for the treatment of recessive dystrophic epidermolysis bullosa.
abstract: Patients with recessive dystrophic epidermolysis bullosa (RDEB) lack functional type VII collagen owing to mutations in the gene COL7A1 and suffer severe blistering and chronic wounds that ultimately lead to infection and development of lethal squamous cell carcinoma. The discovery of induced pluripotent stem cells (iPSCs) and the ability to edit the genome bring the possibility to provide definitive genetic therapy through corrected autologous tissues. We generated patient-derived COL7A1-corrected epithelial keratinocyte sheets for autologous grafting. We demonstrate the utility of sequential reprogramming and adenovirus-associated viral genome editing to generate corrected iPSC banks. iPSC-derived keratinocytes were produced with minimal heterogeneity, and these cells secreted wild-type type VII collagen, resulting in stratified epidermis in vitro in organotypic cultures and in vivo in mice. Sequencing of corrected cell lines before tissue formation revealed heterogeneity of cancer-predisposing mutations, allowing us to select COL7A1-corrected banks with minimal mutational burden for downstream epidermis production. Our results provide a clinical platform to use iPSCs in the treatment of debilitating genodermatoses, such as RDEB.
authors.affiliation: Institute for Stem Cell Biology and Regenerative Medicine, and Department of Pathology, Stanford University, Stanford, CA 94305, USA. Department of Obstetrics and Gynecology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Vittorio
authors.initials: V
authors.lastname: Sebastiano
authors.affiliation: Program in Epithelial Biology, Department of Dermatology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Hanson Hui
authors.initials: HH
authors.lastname: Zhen
authors.forename: Bahareh
authors.initials: B
authors.lastname: Haddad
authors.affiliation: Institute for Stem Cell Biology and Regenerative Medicine, and Department of Pathology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Bahareh Haddad
authors.initials: BH
authors.lastname: Derafshi
authors.affiliation: Institute for Stem Cell Biology and Regenerative Medicine, and Department of Pathology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Elizaveta
authors.initials: E
authors.lastname: Bashkirova
authors.affiliation: Program in Epithelial Biology, Department of Dermatology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Sandra P
authors.initials: SP
authors.lastname: Melo
authors.affiliation: Department of Developmental Biology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Pei
authors.initials: P
authors.lastname: Wang
authors.affiliation: Department of Developmental Biology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Thomas L
authors.initials: TL
authors.lastname: Leung
authors.affiliation: Program in Epithelial Biology, Department of Dermatology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Zurab
authors.initials: Z
authors.lastname: Siprashvili
authors.affiliation: Program in Epithelial Biology, Department of Dermatology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Andrea
authors.initials: A
authors.lastname: Tichy
authors.affiliation: Program in Epithelial Biology, Department of Dermatology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Jiang
authors.initials: J
authors.lastname: Li
authors.affiliation: Program in Epithelial Biology, Department of Dermatology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Ameen
authors.affiliation: Institute for Stem Cell Biology and Regenerative Medicine, and Department of Pathology, Stanford University, Stanford, CA 94305, USA.
authors.forename: John
authors.initials: J
authors.lastname: Hawkins
authors.affiliation: Program in Epithelial Biology, Department of Dermatology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Susie
authors.initials: S
authors.lastname: Lee
authors.affiliation: Program in Epithelial Biology, Department of Dermatology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Lingjie
authors.initials: L
authors.lastname: Li
authors.affiliation: Institute for Regenerative Cures, School of Medicine, University of California, Davis, Sacramento, CA 95817, USA.
authors.forename: Aaron
authors.initials: A
authors.lastname: Schwertschkow
authors.affiliation: Institute for Regenerative Cures, School of Medicine, University of California, Davis, Sacramento, CA 95817, USA.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Bauer
authors.affiliation: Departments of Pediatrics and Genetics, Stanford University, Stanford, CA 94305, USA.
authors.forename: Leszek
authors.initials: L
authors.lastname: Lisowski
authors.affiliation: Departments of Pediatrics and Genetics, Stanford University, Stanford, CA 94305, USA.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Kay
authors.affiliation: Department of Developmental Biology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Seung K
authors.initials: SK
authors.lastname: Kim
authors.affiliation: Program in Epithelial Biology, Department of Dermatology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Alfred T
authors.initials: AT
authors.lastname: Lane
authors.affiliation: Institute for Stem Cell Biology and Regenerative Medicine, and Department of Pathology, Stanford University, Stanford, CA 94305, USA. wernig@stanford.edu oro@stanford.edu.
authors.forename: Marius
authors.initials: M
authors.lastname: Wernig
authors.affiliation: Program in Epithelial Biology, Department of Dermatology, Stanford University, Stanford, CA 94305, USA. wernig@stanford.edu oro@stanford.edu.
authors.forename: Anthony E
authors.initials: AE
authors.lastname: Oro
mesh_terms: D000818:Animals; D001483:Base Sequence; D024143:Collagen Type VII / Q000235:genetics* / Q000627:therapeutic use*; D016108:Epidermolysis Bullosa Dystrophica / Q000235:genetics / Q000628:therapy*; D005808:Genes, Recessive; D020022:Genetic Predisposition to Disease; D015316:Genetic Therapy; D015894:Genome, Human; D059765:Homologous Recombination / Q000235:genetics; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism* / Q000637:transplantation*; D015603:Keratinocytes / Q000473:pathology; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D017422:Sequence Analysis, DNA
chemical_list: C569053:COL7A1 protein, human; D024143:Collagen Type VII