Lentiviral and adeno-associated vectors efficiently transduce mouse T lymphocytes when targeted to murine CD8

Molecular Therapy: Methods & Clinical Development

Mehryad Mataei¹  Christian J. Buchholz²  Jessica Hartmann³  Dirk Grimm³  Annika M. Frank⁴  Dorothee M. Günther⁵  Kathleen Börner⁵  Alexander Michels⁵ 

[1] Fries Lab, Ernst Strüngmann Institute for Neuroscience, 60528 Frankfurt, Germany;German Center for Infection Research (DZIF);Department of Infectious Diseases, Medical Faculty, University of Heidelberg, 69120 Heidelberg, Germany;Division of Medical Biotechnology, Paul-Ehrlich-Institut, 63225 Langen, Germany;Molecular Biotechnology and Gene Therapy, Paul-Ehrlich-Institut, 63225 Langen, Germany;

关键词: splenocytes;    gene therapy;    viral vectors;    chimeric antigen receptor;    CAR;    AAV capsid engineering;   

DOI  :  

来源: DOAJ

【 摘 要 】

Preclinical studies on gene delivery into mouse lymphocytes are often hampered by insufficient activity of lentiviral (LV) and adeno-associated vectors (AAVs) as well as missing tools for cell type selectivity when considering in vivo gene therapy. Here, we selected designed ankyrin repeat proteins (DARPins) binding to murine CD8. The top-performing DARPin was displayed as targeting ligand on both vector systems. When used on engineered measles virus (MV) glycoproteins, the resulting mCD8-LV transduced CD8+ mouse lymphocytes with near-absolute (>99%) selectivity. Despite its lower functional titer, mCD8-LV achieved 4-fold higher gene delivery to CD8+ cells than conventional VSV-LV when added to whole mouse blood. Addition of mCD8-LV encoding a chimeric antigen receptor (CAR) specific for mouse CD19 to splenocytes resulted in elimination of B lymphocytes and lymphoma cells. For display on AAV, the DARPin was inserted into the GH2-GH3 loop of the AAV2 capsid protein VP1, resulting in a DARPin-targeted AAV we termed DART-AAV. Stocks of mCD8-AAV contained similar genome copies as AAV2 but were >20-fold more active in gene delivery in mouse splenocytes, while exhibiting >99% specificity for CD8+ cells. These results suggest that receptor targeting can overcome blocks in transduction of mouse splenocytes.

【 授权许可】

Unknown