Preclinical Corrective Gene Transfer in Xeroderma Pigmentosum Human Skin Stem Cells

Emilie Warrick (1) , Marta Garcia (2) , Corinne Chagnoleau , Odile Chevallier (3, 4) , Valérie Bergoglio (5) , Daniela Sartori , Fulvio Mavilio (6) , Jaime Angulo (7) , Marie-Françoise Avril (8) , Alain Sarasin (9) , Fernando Larcher (10) , Marcela del Rio (10) , Françoise Bernerd (1) , Thierry Magnaldo (9)

1 L'Oréal Recherche - L'Oréal Recherche France
2 CEREGE - Centre européen de recherche et d'enseignement des géosciences de l'environnement
3 ERIGC - Etude des relations instabilité génétique et cancer
4 UMR 8200 - Stabilité Génétique et Oncogenèse
5 IPBS - Institut de pharmacologie et de biologie structurale
6 Généthon
7 Laboratoire de Génétique de la Radiosensibilité, Département de Radiobiologie et Radiopathologie
8 Hôpital Cochin [AP-HP]
9 GC (FRE2939) - Génomes et cancer
10 Epithelial Biomedicine Division [CIEMAT Madrid]

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 (>1040 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.

Domaines: Sciences du Vivant [q-bio]

Emilie Warrick, Marta Garcia, Corinne Chagnoleau, Odile Chevallier, Valérie Bergoglio, et al.. Preclinical Corrective Gene Transfer in Xeroderma Pigmentosum Human Skin Stem Cells. Molecular Therapy, 2012, 20 (4), pp.798-807. ⟨10.1038/mt.2011.233⟩. ⟨hal-02325743⟩