Correction of laminin-5 deficiency in human epidermal stem cells by transcriptionally targeted lentiviral vectors.

Francesca Di Nunzio (1) , Giulietta Maruggi (1) , Stefano Ferrari (2) , Enzo Di Iorio (2) , Valentina Poletti (1) , Marta Garcia (3) , Marcela del Rio (3) , Michele de Luca (1, 2) , Fernando Larcher (3) , Graziella Pellegrini (1, 2) , Fulvio Mavilio (1)

1 UNIMORE - Università degli Studi di Modena e Reggio Emilia = University of Modena and Reggio Emilia
2 The Veneto Eye Bank Foundation, Venice
3 Centre for Biomedical Network Research on Rare Diseases (CIBERER)

Deficiency of the basement membrane component laminin-5 (LAM5) causes junctional epidermolysis bullosa (JEB), a severe and often fatal skin adhesion defect. Autologous transplantation of epidermal stem cells genetically corrected with a Moloney leukemia virus (MLV)-derived retroviral vector reconstitutes LAM5 synthesis, and corrects the adhesion defect in JEB patients. However, MLV-derived vectors have genotoxic characteristics, and are unable to reproduce the physiological, basal layer-restricted expression of LAM5 chains. We have developed an alternative gene transfer strategy based on self-inactivating (SIN) or long terminal repeat (LTR)-modified lentiviral vectors, in which transgene expression is under the control of different combinations of promoter-enhancer elements derived from the keratin-14 (K14) gene. Analysis in human keratinocyte cultures and in fully differentiated skin regenerated onto immunodeficient mice showed that gene expression directed by K14 enhancers is tissue-specific and restricted to the basal layer of the epidermis. Transcriptionally targeted lentiviral vectors efficiently transduced clonogenic stem/progenitor cells derived from a skin biopsy of a JEB patient, restored normal synthesis of LAM5 in cultured keratinocytes, and reconstituted normal adhesion properties in human skin equivalents transplanted onto immunodeficient mice. These vectors are therefore an effective, and potentially more safe, alternative to MLV-based retroviral vectors in gene therapy of JEB.Molecular Therapy (2008) 16 12, 1977-1985 doi:10.1038/mt.2008.204.

Domains: Life Sciences [q-bio]; Biochemistry, Molecular Biology

Francesca Di Nunzio, Giulietta Maruggi, Stefano Ferrari, Enzo Di Iorio, Valentina Poletti, et al.. Correction of laminin-5 deficiency in human epidermal stem cells by transcriptionally targeted lentiviral vectors.. Molecular Therapy, 2008, 16 (12), pp.1977-85. ⟨10.1038/mt.2008.204⟩. ⟨pasteur-01536184⟩