title: High-level transgene expression in human hematopoietic progenitors and differentiated blood lineages after transduction with improved lentiviral vectors.
abstract: Recent experiments point to the great value of lentiviral vectors for the transduction of human hematopoietic stem cells (hHSCs). Vectors used so far, however, have been poorly satisfying in terms of either biosafety or efficiency of transgene expression. Herein is described the results obtained with human immunodeficiency virus-based vectors optimized in both of these aspects. It is thus shown that vectors containing the EF1alpha and, to a lesser extent, the phosphoglycerate kinase (PGK) promoter, govern high-level gene expression in human hematopoietic progenitors as well as derived hematopoietic lineages of therapeutic relevance, such as erythrocytes, granulocytes, monocytes, dendritic cells, and megakaryocytes. EF1alpha promoter-containing lentiviral vectors can also induce strong transgene expression in primary T lymphocytes isolated from peripheral blood. A self-inactivating design did not affect the performance of EF1alpha promoter-based vectors but significantly reduced expression from the PGK promoter. This negative effect could nevertheless be largely rescued by inserting the post-transcriptional regulatory element of woodchuck hepatitis virus upstream of the vector 3' long terminal repeat. These results have important practical implications for the genetic treatment of lymphohematologic disorders as well as for the study of hematopoiesis via the lentivector-mediated modification of hHSCs.
authors.affiliation: Department of Genetics and Microbiology and Division of Hematology, Faculty of Medicine, Geneva, Switzerland.
authors.forename: P
authors.initials: P
authors.lastname: Salmon
authors.forename: V
authors.initials: V
authors.lastname: Kindler
authors.forename: O
authors.initials: O
authors.lastname: Ducrey
authors.forename: B
authors.initials: B
authors.lastname: Chapuis
authors.forename: R H
authors.initials: RH
authors.lastname: Zubler
authors.forename: D
authors.initials: D
authors.lastname: Trono
mesh_terms: D018952:Antigens, CD34 / Q000097:blood; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D005312:Fetal Blood / Q000166:cytology / Q000378:metabolism; D005434:Flow Cytometry; D015967:Gene Expression Regulation, Viral; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000592:standards*; D006678:HIV / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D017919:Hepatitis B Virus, Woodchuck / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008979:Moloney murine leukemia virus / Q000235:genetics; D020648:Peptide Elongation Factor 1 / Q000235:genetics; D010735:Phosphoglycerate Kinase / Q000235:genetics; D011401:Promoter Regions, Genetic; D012323:RNA Processing, Post-Transcriptional; D013601:T-Lymphocytes / Q000378:metabolism; D014161:Transduction, Genetic; D019076:Transgenes / Q000235:genetics*
chemical_list: D018952:Antigens, CD34; D004279:DNA, Viral; D020648:Peptide Elongation Factor 1; D010735:Phosphoglycerate Kinase