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Reduced Uptake of Oxidized Low-Density Lipoprotein by Macrophages Using Multiple Aptamer Combinations
Soemwit Khongwichit1,2, Piyawut Swangphon1, Aekkaraj Nualla-Ong2,3,4
1Faculty of Medical Technology, Prince of Songkla University, Songkhla 90110, Thailand.
Abstract:
The accumulation of oxidized low-density lipoprotein (oxLDL) in macrophages leads to the formation of foam cells and atherosclerosis development. Reducing the uptake of oxLDL in macrophages decreases the incidence and progression of atherosclerosis. Four distinct single-strand DNA sequences, namely, AP07, AP11, AP25, and AP29, were selected that demonstrated specific binding to distinct regions of oxidized apolipoprotein B100 (apoB100; the protein component of oxLDL) with low HDOCK scores. These four DNA sequences were combined to generate aptamers that selectively bound to labeled Dil-oxLDL, and were subsequently added to murine RAW 264.7 macrophages to test their inhibitory effects using fluorescence spectrometry. The four combined aptamers at 10 μM reduced oxLDL uptake by 79 ± 4% compared to that of the untreated aptamer group. Flow cytometry data demonstrated that macrophages treated with aptamers reached only 32.6% of the Dil-oxLDL signal, a 50% reduction in fluorescence emission relative to that of the untreated group (64.4% Dil-oxLDL signal). Binding the four combined aptamers to the oxLDL surface disrupted the interaction between oxLDL and CD36 via cyclic voltammetry, effectively decreasing the level of uptake of oxLDL by macrophages. Results suggested that these aptamers could be used as alternative compounds to prevent the formation of foam cells, hence providing antiatherosclerosis activity.
Insights
New DNA aptamers significantly inhibit oxidized LDL uptake by macrophages, reducing foam cell formation and offering potential anti-atherosclerosis activity. This discovery could lead to novel treatments for preventing atherosclerosis progression.
Area of Science:
- Biochemistry
- Molecular Biology
- Cardiovascular Research
Background:
- Oxidized low-density lipoprotein (oxLDL) accumulation in macrophages drives foam cell formation and atherosclerosis.
- Reducing oxLDL uptake by macrophages is a key strategy to decrease atherosclerosis incidence and progression.
Purpose of the Study:
- To develop and evaluate novel DNA aptamers for inhibiting oxLDL uptake in macrophages.
- To assess the potential of these aptamers as anti-atherosclerosis therapeutic agents.
Main Methods:
- Selection of four DNA aptamers (AP07, AP11, AP25, AP29) with specific binding to oxidized apolipoprotein B100 (apoB100).
- Combination of aptamers to form a complex that selectively binds to Dil-oxLDL.
- Assessment of aptamer efficacy in reducing oxLDL uptake in murine RAW 264.7 macrophages using fluorescence spectrometry and flow cytometry.
- Investigation of the mechanism of action, including disruption of oxLDL-CD36 interaction via cyclic voltammetry.
Main Results:
- The combined aptamers at 10 μM reduced oxLDL uptake by 79 ± 4% in macrophages.
- Flow cytometry showed a 50% reduction in Dil-oxLDL signal in aptamer-treated macrophages compared to controls.
- Cyclic voltammetry confirmed that aptamers disrupt the interaction between oxLDL and the CD36 scavenger receptor.
Conclusions:
- The developed DNA aptamers effectively inhibit oxLDL uptake by macrophages.
- These aptamers demonstrate potential as therapeutic compounds to prevent foam cell formation and exert anti-atherosclerosis activity.
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