A macrophage-specific synthetic promoter for therapeutic application of adiponectin

Abstract

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 p47phox 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 p47phox promoter element, was successfully developed to target macrophage, and macrophage-specific introduction of APN under SP146-C1 was shown to ameliorate the atherosclerotic pathology.

MDM2 E3 ligase-mediated ubiquitination and degradation of HDAC1 in vascular calcification

“…The atherosclerosis model was generated 56 . Subtotal occlusion of the left carotid artery was carried out in male ApoE KO mice 57 .…”

PKA regulatory IIα subunit is essential for PGD2-mediated resolution of inflammation

“…DP1 flox/flox littermates (WT) served as experimental controls. Macrophage-specific DP1 transgenic (Mac-DP1 Tg) mice were generated by Caygen Biosciences on the C57BL/6 genetic background using macrophage-specific synthetic promoter 146 (SP146; Kang et al, 2014 ). PRKAR2A −/− mice were maintained on C57BL/6 genetic background.…”

Toll-Like Receptor 8 Agonist Strengthens the Protective Efficacy of ESAT-6 Immunization to Mycobacterium tuberculosis Infection

“…All mice were housed under SPF-level conditions. TLR8 human cDNA ORF (NM_138636) (OriGene) with a macrophage-specific synthetic promoter SP146+ P47 ( 19 , 20 ) was transferred into C57BL/6 mice to acquire the transgenic mice. Ifnar1 −/− mice ( 21 ) were crossed with TLR8 transgenic mice.…”