A capsid-modified adenovirus vector devoid of all viral genes: assessment of transduction and toxicity in human hematopoietic cells.
Inefficient gene transfer has limited the success of gene therapy in the hematopoietic system. Here we develop a novel chimeric adenovirus (Ad) vector containing Ad serotype 11 fiber-modified capsids and E1/E3 deleted viral genomes (Ad5/11) or genomes devoid of all viral genes (DeltaAd5/11). The capsid-modified vectors transduced human hematopoietic cells more efficiently than the unmodified Ad5-based vector. The absence of viral genes from the DeltaAd5/11 vector allowed for transduction without the associated toxicity seen with the first-generation E1/E3 deleted vector. Chimeric vectors were used for transient expression of the ecotropic retrovirus receptor (ecoR) in Mo7e cells (a CD34-positive, c-Kit-positive, growth-factor-dependent human cell line) as a model for human hematopoietic progenitor cells. Expression of ecoR conferred susceptibility to subsequent retroviral transduction. The DeltaAd5/11 vector used to express ecoR allowed for expansion of retrovirally transduced cells, whereas transduction with the first-generation Ad5/11 vector resulted in cytotoxicity and, over time, loss of cells expressing the retrovirus-vector-derived transgene.
Stecher
H
H
Division of Medical Genetics, Department of Medicine, University of Washington, Seattle, WA 98195, USA.
Shayakhmetov
D M
DM
Stamatoyannopoulos
G
G
Lieber
A
A
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D018952:Antigens, CD34 / Q000096:biosynthesis; D017209:Apoptosis; D002213:Capsid / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D017353:Gene Deletion; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000633:toxicity; D016679:Genome, Viral; D006412:Hematopoietic Stem Cells; D006801:Humans; D051381:Rats; D014161:Transduction, Genetic; D019076:Transgenes
D018952:Antigens, CD34