Gene transfer into human cord blood−derived CD34+ cells by adeno-associated viral vectors

Schuhmann, Natascha K.; Pozzoli, Ombretta; Sallach, Jessica; Huber, Anke; Avitabile, Daniele; Perabo, Luca; Rappl, Gunter; Capogrossi, Maurizio C.; Hallek, Michael; Pesce, Maurizio; Büning, Hildegard

doi:10.1016/j.exphem.2010.04.016

« Previous Next »Experimental Hematology
Volume 38, Issue 9 , Pages 707-717, September 2010

Gene transfer into human cord blood−derived CD34⁺ cells by adeno-associated viral vectors

Natascha K. Schuhmann
- Department I of Internal Medicine, University of Cologne, Cologne, Germany
- Drs. Schuhmann and Pozzoli contributed equally to this work, as did Drs. Pesce and Büning.
Ombretta Pozzoli
- Laboratorio di Biologia Vascolare e Medicina Rigenerativa, Centro Cardiologico Monzino−IRCCS, Milan, Italy
- Drs. Schuhmann and Pozzoli contributed equally to this work, as did Drs. Pesce and Büning.
Jessica Sallach
- Department I of Internal Medicine, University of Cologne, Cologne, Germany
- Center for Molecular Medicine Cologne (ZMMK), University of Cologne, Cologne, Germany
Anke Huber
- Department I of Internal Medicine, University of Cologne, Cologne, Germany
- Center for Molecular Medicine Cologne (ZMMK), University of Cologne, Cologne, Germany
Daniele Avitabile
- Laboratorio di Patologia Vascolare, Istituto Dermopatico dell' Immacolata−IRCCS, Rome, Italy
Luca Perabo
- Department I of Internal Medicine, University of Cologne, Cologne, Germany
Gunter Rappl
- Department I of Internal Medicine, University of Cologne, Cologne, Germany
- Center for Molecular Medicine Cologne (ZMMK), University of Cologne, Cologne, Germany
Maurizio C. Capogrossi
- Laboratorio di Patologia Vascolare, Istituto Dermopatico dell' Immacolata−IRCCS, Rome, Italy
Michael Hallek
- Department I of Internal Medicine, University of Cologne, Cologne, Germany
- Center for Molecular Medicine Cologne (ZMMK), University of Cologne, Cologne, Germany
Maurizio Pesce
- Laboratorio di Biologia Vascolare e Medicina Rigenerativa, Centro Cardiologico Monzino−IRCCS, Milan, Italy
- Drs. Schuhmann and Pozzoli contributed equally to this work, as did Drs. Pesce and Büning.
Hildegard Büning
- Department I of Internal Medicine, University of Cologne, Cologne, Germany
- Center for Molecular Medicine Cologne (ZMMK), University of Cologne, Cologne, Germany
- Drs. Schuhmann and Pozzoli contributed equally to this work, as did Drs. Pesce and Büning.
- Offprint requests to: Hildegard Büning, Ph.D., University of Cologne, Department I of Internal Medicine, Center for Molecular Medicine Cologne (ZMMK), ZMMK Research Building, Robert-Koch-Str. 21, Cologne 50937, Germany

Received 21 January 2009; received in revised form 8 April 2010; accepted 27 April 2010. published online 05 May 2010.

Objective

Bone marrow−derived CD34⁺ cells are currently used in clinical trials in patients with ischemic heart disease. An option to enhance activity of injected progenitors may be offered by genetic engineering of progenitor cells with angiogenic growth factors. Recombinant adeno-associated viral vectors (rAAV) have emerged as a leading gene transfer systems. In contrast to other vector systems in use for genetic engineering of CD34⁺ cells, rAAV-mediated gene expression does not depend on vector integration. This is relevant for application in regenerative medicine of ischemic tissues, where transient transgene expression is likely sufficient to achieve therapeutic benefits.

Materials and Methods

We compared three different human AAV serotypes, packaged as pseudotypes by a helper virus-free production method, for their transduction efficiency in human cord blood−derived CD34⁺ cells. We further assessed the impact of vector genome conformation, of α_vβ₅ and α₅β₁ integrin availability and of the transcription-modulating drugs retinoic acid and Trichostatin A on rAAV-mediated human CD34⁺ cell transduction.

Results

We provide, for the first time, evidence that hCD34⁺ cells can be reproducibly transduced with high efficiency by self-complementary rAAV2 without inducing cytotoxicity or interfering with their differentiation potential. We further show the involvement of α₅β₁ integrin as a crucial AAV2 internalization receptor and a function for transcription-modulating drugs in enhancing rAAV-mediated transgene expression.