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Persistent mismatch repair deficiency following targeted correction of hMLH1

Cancer Gene Therapy volume 14, pages 98–104 (2007)Cite this article

Abstract

The use of gene therapy to correct mutated or lost gene function for the treatment of human cancers has been an active, yet problematic area of biomedical research. Many technical difficulties, including efficient tissue-specific delivery, integration site specificity and general toxicity, are being addressed. Little is known, however, about the genetic and phenotypic stability that accompanies a successful gene-specific targeting event in a cancer cell. This question was addressed following the creation of a colon cancer cell line in which a mutated hMLH1 gene was corrected via targeted homologous recombination. This correction resulted in the expression of wild-type hMLH1 protein, restoration of the hPMS2 protein and mismatch repair (MMR) proficiency. One of two hMLH1-corrected clones, however, was found to retain defects in MMR activity. These cells continued to express the corrected hMLH1 protein, but had lost expression of another MMR protein, hMSH6. DNA sequence analysis of the hMSH6 gene revealed biallelic expansions of a cytosine repeat region in exon 5 that result in frameshifts leading to premature stop codons. These findings suggest that, similar to acquired drug resistance, the presence of genetically heterogeneous cancer cell populations or acquisition of compensatory mutations can result in ‘resistance’ to gene replacement therapy.

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Acknowledgements

We thank Drs Bert Vogelstein and Joseph Jiricny for helpful advice and the critical reading of the manuscript and Drs James Carney and Teresa Wilson for helpful discussions. This work was funded by the Maryland Cigarette Restitution Fund, The Flight Attendant's Medical Research Institute (FAMRI), NIH 1R01CA109274-01A2, The Department of Defense Breast Cancer Research Program (DAMD17-03-1-0241) and the Avon Foundation. BHP is an Avon Scholar for Breast Cancer Research and also receives support from The V Foundation for Cancer Research.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, University of Maryland Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, MD, USA

    M B Weiss, M I Vitolo & K E Bachman

  2. University of Maryland Graduate Program in Molecular Medicine, Baltimore, MD, USA

    M B Weiss & K E Bachman

  3. Institute of Molecular Cancer Research, University of Zurich, Zurich, Switzerland

    K Baerenfaller & G Marra

  4. Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, The Johns Hopkins University School of Medicine, Baltimore, MD, USA

    B H Park

  5. Department of Radiation Oncology, University of Maryland Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, MD, USA

    K E Bachman

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  1. M B Weiss
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Correspondence to K E Bachman.

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Weiss, M., Vitolo, M., Baerenfaller, K. et al. Persistent mismatch repair deficiency following targeted correction of hMLH1. Cancer Gene Ther 14, 98–104 (2007). https://doi.org/10.1038/sj.cgt.7700997

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