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

[junctional epidermolysis bullosa]

Deficiency of the basement membrane component laminin-5 (LAM 5 ) 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 LAM 5 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 LAM 5 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 (K 14 ) gene . Analysis in human keratinocyte cultures and in fully differentiated skin regenerated onto immunodeficient mice showed that gene expression directed by K 14 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 LAM 5 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 .

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Diseases presenting "long terminal repeat" symptom

junctional epidermolysis bullosa

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