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Specific gene transfer to neurons, endothelial cells and hematopoietic progenitors with lentiviral vectors

Nature Methods volume 7, pages 929–935 (2010)Cite this article

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Abstract

We present a flexible and highly specific targeting method for lentiviral vectors based on single-chain antibodies recognizing cell-surface antigens. We generated lentiviral vectors specific for human CD105+ endothelial cells, human CD133+ hematopoietic progenitors and mouse GluA-expressing neurons. Lentiviral vectors specific for CD105 or for CD20 transduced their target cells as efficiently as VSV-G pseudotyped vectors but discriminated between endothelial cells and lymphocytes in mixed cultures. CD133-targeted vectors transduced CD133+ cultured hematopoietic progenitor cells more efficiently than VSV-G pseudotyped vectors, resulting in stable long-term transduction. Lentiviral vectors targeted to the glutamate receptor subunits GluA2 and GluA4 exhibited more than 94% specificity for neurons in cerebellar cultures and when injected into the adult mouse brain. We observed neuron-specific gene modification upon transfer of the Cre recombinase gene into the hippocampus of reporter mice. This approach allowed targeted gene transfer to many cell types of interest with an unprecedented degree of specificity.

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Figure 1: Characterization of targeted lentiviral vectors.
Figure 2: Selective and efficient transduction of primary endothelial cells by CD105-LV.
Figure 3: Transduction of primary human HPCs via CD133.
Figure 4: Specific gene transfer to primary neurons using GluA2/4-LV.
Figure 5: In vivo application of GluA2/4-LV.

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Acknowledgements

This work was supported by grants from the Priority Programme “Mechanisms of gene vector entry and persistence” of the Deutsche Forschungsgemeinschaft to C.J.B. and K.C. and from the 7th European Community programme project Persisting Transgenesis (Persist) to C.J.B. U.C.M. was supported by grants of the Deutsche Forschungsgemeinschaft (MU 1457/8-1) and the “Nationales Genomforschungsnetz” (01GS08128). T.A. is supported by the graduate study program GK1172 Biologicals of the Goethe University Frankfurt AM.

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Author notes

  1. Brigitte Anliker, Tobias Abel and Sabrina Kneissl: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Medical Biotechnology, Paul-Ehrlich-Institut, Langen, Germany

    Brigitte Anliker, Tobias Abel, Sabrina Kneissl, Julia Brynza, Irene C Schneider, Robert C Münch, Klaus Cichutek & Christian J Buchholz

  2. Department of Pathobiology, Institute of Virology, University of Veterinary Medicine, Vienna, Austria

    Juraj Hlavaty, Helga Petznek & Christine Hohenadl

  3. Institute for Clinical Neurobiology, University Hospital of Neurology, Heidelberg, Germany

    Antonio Caputi & Hannah Monyer

  4. Institute of Cell Biology and Immunology, University of Stuttgart, Stuttgart, Germany

    Roland E Kontermann

  5. Pediatric Hematology and Oncology, University Hospital, Frankfurt, Germany

    Ulrike Koehl

  6. Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany

    Ian C D Johnston

  7. Department of Biosciences, Division of Biochemistry, University of Helsinki, Finland

    Kari Keinänen

  8. Institute for Pharmacy and Molecular Biotechnology, University of Heidelberg, Germany

    Ulrike C Müller

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  1. Brigitte Anliker
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Contributions

B.A., T.A. and S.K. designed and performed experiments and contributed to writing of the manuscript. J.H., A.C., J.B., I.C.S., R.C.M. and H.P. performed experiments. R.E.K., U.K., I.C.D.J. and K.K. contributed protocols and reagents. C.H. and H.M. supervised work. U.C.M. supervised work and contributed to writing of the manuscript. K.C. acquired grants. C.J.B. conceived and designed the study, acquired grants, supervised work and wrote the manuscript.

Corresponding author

Correspondence to Christian J Buchholz.

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Competing interests

I.C.D.J. is an employee of Miltenyi Biotec GmbH. S.K., K.C. and C.J.B. are listed as inventors in an international Patent Cooperation Treaty European patent application (PCT/EP2007/008384) assigned to the Paul Ehrlich Institut, which includes as claims the generation of targeted lentiviral vectors.

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Anliker, B., Abel, T., Kneissl, S. et al. Specific gene transfer to neurons, endothelial cells and hematopoietic progenitors with lentiviral vectors. Nat Methods 7, 929–935 (2010). https://doi.org/10.1038/nmeth.1514

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