Preclinical correction of human Fanconi anemia complementation group A bone marrow cells using a safety-modified lentiviral vector

P. S. Becker, J. A. Taylor, G. D. Trobridge, X. Zhao, B. C. Beard, S. Chien, J. Adair, D. B. Kohn, J. E. Wagner, A. Shimamura, H. P. Kiem

Research output: Contribution to journalArticlepeer-review

34 Scopus citations


One of the major hurdles for the development of gene therapy for Fanconi anemia (FA) is the increased sensitivity of FA stem cells to free radical-induced DNA damage during ex vivo culture and manipulation. To minimize this damage, we have developed a brief transduction procedure for lentivirus vector-mediated transduction of hematopoietic progenitor cells from patients with Fanconi anemia complementation group A (FANCA). The lentiviral vector FancA-sW contains the phosphoglycerate kinase promoter, the FANCA cDNA, and a synthetic, safety-modified woodchuck post transcriptional regulatory element (sW). Bone marrow mononuclear cells or purified CD34+cells from patients with FANCA were transduced in an overnight culture on recombinant fibronectin peptide CH-296, in low (5%) oxygen, with the reducing agent, N-acetyl-L-cysteine (NAC), and a combination of growth factors, granulocyte colony-stimulating factor (G-CSF), Flt3 ligand, stem cell factor, and thrombopoietin. Transduced cells plated in methylcellulose in hypoxia with NAC showed increased colony formation compared with 21% oxygen without NAC (P<0.03), showed increased resistance to mitomycin C compared with green fluorescent protein (GFP) vector-transduced controls (P<0.007), and increased survival. Thus, combining short transduction and reducing oxidative stress may enhance the viability and engraftment of gene-corrected cells in patients with FANCA.

Original languageEnglish (US)
Pages (from-to)1244-1252
Number of pages9
JournalGene therapy
Issue number10
StatePublished - Oct 2010

Bibliographical note

Funding Information:
This work was supported by NIH Grants HL085693, DK56465 and DK47754 to HPK. We thank Helen Crawford and Bonnie Larson for help with the preparation of the article. HPK is a Molecular Medicine Investigator and the recipient of the José Carreras/ E. Donnall Thomas Endowed Chair for Cancer Research. We received grant support from National Institutes of Health.


  • Hhypoxia
  • gene therapy
  • mitomycin C
  • reducing agent


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