Evaluation of macrophage-specific promoters using lentiviral delivery in mice

• Published in: Gene therapy Vol. 19; no. 11; pp. 1041 - 1047
• Main Authors: Levin, M C, Lidberg, U, Jirholt, P, Adiels, M, Wramstedt, A, Gustafsson, K, Greaves, D R, Li, S, Fazio, S, Linton, M F, Olofsson, S-O, Borén, J, Gjertsson, I
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01-11-2012; Nature Publishing Group
• Subjects: 631/326/596/2561; 631/61/2300; 692/698/690/292; 692/700/565/201; Analysis; Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Animals; Applied cell therapy and gene therapy; Biological and medical sciences; Biomedical and Life Sciences; Biomedicine; Biotechnology; Cell Biology; Cell Line; Cell lines; Clinical Medicine; Expression vectors; Flow cytometry; Fundamental and applied biological sciences. Psychology; Gene Dosage; Gene Expression; Gene Order; Gene Therapy; Gene transfer; Genetic Therapy; Genetic Vectors - genetics; Green fluorescent protein; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Health aspects; Health. Pharmaceutical industry; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hematopoietic Stem Cells - metabolism; Human Genetics; Humans; Industrial applications and implications. Economical aspects; Klinisk medicin; Lentivirus; Lentivirus - genetics; Macrophages; Macrophages - metabolism; Medical research; Medical sciences; Mice; MSP protein; Nanotechnology; Organ Specificity - genetics; original-article; Physiological aspects; Promoter Regions, Genetic; Promoters; Promoters (Genetics); Stem cell transplantation; Stem cells; Transcription factors; Transduction, Genetic; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; Transgenes; Viruses; United States; Sweden; macrophage; promoter; lentivirus; Virus; Vertebrata; Mammalia; Mouse; Rodentia; Retroviridae; Gene therapy; Lentivirus; Macrophage
• ISSN: 0969-7128; 1476-5462; 1476-5462
• DOI: 10.1038/gt.2011.195

In gene therapy, tissue-specific promoters are useful tools to direct transgene expression and improve efficiency and safety. Macrophage-specific promoters (MSPs) have previously been published using different delivery systems. In this study, we evaluated five different MSPs fused with green fluorescent protein (GFP) to delineate the one with highest specificity using lentiviral delivery. We compared three variants of the CD68 promoter (full length, the 343-bp proximal part and the 150-bp proximal part) and two variants (in forward and reverse orientation) of a previously characterized synthetic promoter derived from elements of transcription factor genes. We transduced a number of cell lines and primary cells
in vitro
. In addition, hematopoietic stem cells were transduced with MSPs and transferred into lethally irradiated recipient mice. Fluorescence activated cell sorting analysis was performed to determine the GFP expression in different cell populations both
in vitro
and
in vivo
. We showed that MSPs can efficiently be used for lentiviral gene delivery and that the 150-bp proximal part of the CD68 promoter provides primarily macrophage-specific expression of GFP. We propose that this is the best currently available MSP to use for directing transgene expression to macrophage populations
in vivo
using lentiviral vectors.