Preclinical corrective gene transfer in Xeroderma pigmentosum human skin stem cells Articles uri icon

publication date

  • April 2012

start page

  • 798

end page

  • 807

issue

  • 4

volume

  • 20

international standard serial number (ISSN)

  • 1525-0016

electronic international standard serial number (EISSN)

  • 1525-0024

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.

keywords

  • dystrioguc epidermolysis-bullosa; human epidermal-keratinocytes; nucleotide excision-repair; long-term engraftement; in-vivo assessment; group-c protein; dna-repair; retroviral vectors ; ; therapy ; ; transduction