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In vivo selection using a cell-growth switch

Abstract

A major obstacle to stem-cell gene therapy rests in the inability to deliver a gene into a therapeutically relevant fraction of stem cells1. One way to circumvent this obstacle is to use selection. Vectors containing two linked genes serve as the basis for selection, with one gene encoding a selectable product and the other, a therapeutic protein. Applying selection in vivo has the potential to bring a minor population of genetically corrected cells into the therapeutic range. But strategies for achieving in vivo selection have traditionally relied on genes that confer resistance to cytotoxic drugs2,3 and are encumbered by toxicity. Here we describe a new system for in vivo selection that uses a ‘cell-growth switch’, allowing a minor population of genetically modified cells to be inducibly amplified, thereby averting the risks associated with cytotoxic drugs. This system provides a general platform for conditionally expanding genetically modified cell populations in vivo, and may have widespread applications in gene and cell therapy.

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Figure 1: Dose-finding study for AP20187. Results for marrow (a) and spleen (b) are shown.
Figure 2: Southern-blot analysis of spleen DNA.
Figure 3: Kinetics of the haematological response to CID treatment.
Figure 4: CID-treatment does not exhaust genetically modified stem cells.

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Acknowledgements

We thank T. Clackson, D. Thomis, F. Cerasoli and M. Gilman for supplying AP20187; K. Humphries for providing the MGIN producer cell line; R. Hawley for providing the MSCVneo and MGIN constructs; D. Persons and J. Ieremia for technical advice and assistance; and T. Clackson, T. Papayannopoulou, J. Abkowitz, K. Kaushansky, P. Martin, D. Russell and G. Stamatoyannopoulos for comments and suggestions regarding the manuscript. This work was supported by grants 5R01DK52997, 1R01DK57525, 2P01HL53750 and 2P01DK47754 from the National Institutes of Health, an American Society of Hematology Junior Faculty Scholar Award and an award from the Fanconi Anemia Research Fund.

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Correspondence to C. Anthony Blau.

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Jin, L., Zeng, H., Chien, S. et al. In vivo selection using a cell-growth switch. Nat Genet 26, 64–66 (2000). https://doi.org/10.1038/79194

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