title: Preclinical corrective gene transfer in xeroderma pigmentosum human skin stem cells.
abstract: Xeroderma pigmentosum (XP) is a devastating disease associated with dramatic skin cancer proneness. XP cells are deficient in nucleotide excision repair (NER) of bulky DNA adducts including ultraviolet (UV)-induced mutagenic lesions. Approaches of corrective gene transfer in NER-deficient keratinocyte stem cells hold great hope for the long-term treatment of XP patients. To face this challenge, we developed a retrovirus-based strategy to safely transduce the wild-type XPC gene into clonogenic human primary XP-C keratinocytes. De novo expression of XPC was maintained in both mass population and derived independent candidate stem cells (holoclones) after more than 130 population doublings (PD) in culture upon serial propagation (>10(40) cells). Analyses of retrovirus integration sequences in isolated keratinocyte stem cells suggested the absence of adverse effects such as oncogenic activation or clonal expansion. Furthermore, corrected XP-C keratinocytes exhibited full NER capacity as well as normal features of epidermal differentiation in both organotypic skin cultures and in a preclinical murine model of human skin regeneration in vivo. The achievement of a long-term genetic correction of XP-C epidermal stem cells constitutes the first preclinical model of ex vivo gene therapy for XP-C patients.
authors: Laboratory of genomes biology and pathologies, CNRS UMR/INSERM, Nice, France.
authors: Emilie
authors: E
authors: Warrick
authors: Marta
authors: M
authors: Garcia
authors: Corinne
authors: C
authors: Chagnoleau
authors: Odile
authors: O
authors: Chevallier
authors: Valérie
authors: V
authors: Bergoglio
authors: Daniela
authors: D
authors: Sartori
authors: Fulvio
authors: F
authors: Mavilio
authors: Jaime F
authors: JF
authors: Angulo
authors: Marie-Françoise
authors: MF
authors: Avril
authors: Alain
authors: A
authors: Sarasin
authors: Fernando
authors: F
authors: Larcher
authors: Marcela
authors: M
authors: Del Rio
authors: Françoise
authors: F
authors: Bernerd
authors: Thierry
authors: T
authors: Magnaldo
mesh_terms: D015139:Blotting, Southern; D015153:Blotting, Western; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism; D005434:Flow Cytometry; D015316:Genetic Therapy; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012867:Skin / Q000166:cytology* / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*; D014983:Xeroderma Pigmentosum / Q000378:metabolism / Q000628:therapy*
chemical_list: D004268:DNA-Binding Proteins; C086984:XPC protein, human