Long-term engraftment of single genetically modified human epidermal holoclones enables safety pre-assessment of cutaneous gene therapy

Fernando Larcher, Elena Dellambra, Laura Rico, Sergio Bondanza, Rodolfo Murillas, Claudia Cattoglio, Fulvio Mavilio, José L. Jorcano, Giovanna Zambruno, Marcela Del Rio

Research output: Contribution to journalArticlepeer-review

Abstract

Predicting the risks of permanent gene therapy approaches involving the use of integrative gene-targeting vectors has become a critical issue after the unfortunate episode of a clinical trial in children with X-linked severe combined immunodeficiency (X-SCID). Safety pre-assessment of single isolated gene-targeted stem cells or their derivative clones able to regenerate their tissue of origin would be a major asset in addressing untoward gene therapy effects in advance. Human epidermal stem cells, which have extensive proliferative potential in vitro, theoretically offer such a possibility as a method of assessment. By means of optimized organotypic culture and grafting methods, we demonstrate the long-term in vivo regenerative capacity of single gene-targeted human epidermal stem cell clones (holoclones). Both histopathological analysis of holoclone-derived grafts in immunodeficient mice and retroviral insertion site mapping performed in the holoclone in vitro and after grafting provide proof of the feasibility of pre-assessing genotoxicity risks in isolated stem cells before transplantation into patients. Our results provide an experimental basis for previously untested assumptions about the in vivo behavior of epidermal stem cells prospectively isolated in vitro and pave the way for a safer approach to cutaneous gene therapy.

Original languageEnglish
Pages (from-to)1670-1676
Number of pages7
JournalMolecular Therapy
Volume15
Issue number9
DOIs
Publication statusPublished - Sep 2007

ASJC Scopus subject areas

  • Molecular Biology

Fingerprint Dive into the research topics of 'Long-term engraftment of single genetically modified human epidermal holoclones enables safety pre-assessment of cutaneous gene therapy'. Together they form a unique fingerprint.

Cite this