A macrophage-specific synthetic promoter for therapeutic application of adiponectin.
Foam cell formation from macrophage is a major cause of atherosclerosis. An efficient macrophage-specific promoter is required for the targeting to macrophages. In this study, we develop a macrophage-specific synthetic promoter for the therapeutic application of adiponectin (APN), an antiatherogenic gene. Synthetic promoter-146 (SP146), registered on the NCBI website (http://www.ncbi.nlm.nih.gov/nuccore/DQ107383), was tested for promoter activities in two non-macrophage cell lines (293 T, HeLa) and a macrophage cell line (RAW264.7, bone marrow-derived macrophages). To enforce macrophage specificity, partial elements of p47(phox) including the PU.1 site with various lengths (-C1, -C2 and -C3) were inserted next to the synthetic promoters. SP146-C1 showed the highest specificity and efficacy in RAW264.7 cells and was selected for development of an APN-carrying macrophage-specific promoter. Green fluorescent protein (GFP)- or APN-expressing lentivirus under SP146-C1 (Lenti-SP-GFP or Lenti-SP-APN, respectively) showed the highest expression efficacy in RAW264.7 cells compared with the non-macrophage cell lines. APN overexpression in RAW264.7 cells successfully inhibited intracellular lipid accumulation, and atherosclerotic lesions and lipid accumulation were significantly reduced by Lenti-SP-APN in ApoE-/- atherosclerosis mice. In conclusion, the synthetic promoter SP146-C1, combined with a p47(phox) promoter element, was successfully developed to target macrophage, and macrophage-specific introduction of APN under SP146-C1 was shown to ameliorate the atherosclerotic pathology.
1] Research Laboratory of Cardiovascular Regeneration, Chonnam National University Hospital, Gwangju, Korea [2] Center of Molecular Medicine, Graduate School, Chonnam National University, Gwangju, Korea.
W S
WS
Kang
1] Research Laboratory of Cardiovascular Regeneration, Chonnam National University Hospital, Gwangju, Korea [2] Heart Research Center, Chonnam National University Hospital, Gwangju, Korea.
J S
JS
Kwon
Research Laboratory of Cardiovascular Regeneration, Chonnam National University Hospital, Gwangju, Korea.
H B
HB
Kim
Research Laboratory of Cardiovascular Regeneration, Chonnam National University Hospital, Gwangju, Korea.
H-Y
HY
Jeong
Research Laboratory of Cardiovascular Regeneration, Chonnam National University Hospital, Gwangju, Korea.
H J
HJ
Kang
1] Heart Research Center, Chonnam National University Hospital, Gwangju, Korea [2] Department of Cardiology, Chonnam National University Hospital, Gwangju, Korea.
M H
MH
Jeong
Department of Cardiology, Chonnam National University Hospital, Gwangju, Korea.
J G
JG
Cho
Department of Cardiology, Chonnam National University Hospital, Gwangju, Korea.
J C
JC
Park
1] Research Laboratory of Cardiovascular Regeneration, Chonnam National University Hospital, Gwangju, Korea [2] Heart Research Center, Chonnam National University Hospital, Gwangju, Korea.
Y S
YS
Kim
1] Research Laboratory of Cardiovascular Regeneration, Chonnam National University Hospital, Gwangju, Korea [2] Heart Research Center, Chonnam National University Hospital, Gwangju, Korea [3] Department of Cardiology, Chonnam National University Hospital, Gwangju, Korea.
Y
Y
Ahn
D052242:Adiponectin / Q000235:genetics* / Q000627:therapeutic use; D000818:Animals; D050197:Atherosclerosis / Q000235:genetics* / Q000473:pathology / Q000628:therapy; D005487:Foam Cells / Q000378:metabolism / Q000473:pathology; D015316:Genetic Therapy; D006367:HeLa Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic
D052242:Adiponectin