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Local gene silencing of monocyte chemoattractant protein-1 prevents vulnerable plaque disruption in apolipoprotein
Xiao Ling Liu1, Peng Fei Zhang, Shi Fang Ding
1Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Qilu Hospital, Shandong University, Jinan, Shandong, China.
Abstract:
Monocyte chemoattractant protein-1 (MCP-1), a CC chemokine (CCL2), has been demonstrated to play important roles in atherosclerosis and becoming an important therapeutic target for atherosclerosis. The present study was undertaken to test the hypothesis that local RNAi of MCP-1 by site-specific delivery of adenovirus-mediated small hairpin RNA (shRNA) may enhance plaque stability and prevent plaque disruption in ApoE-/- mice. We designed an adenovirus-mediated shRNA against mouse MCP-1 (rAd5-MCP-1-shRNA). Male apolipoprotein E-knockout (ApoE-/-) mice (n = 120) were fed a high-fat diet and vulnerable plaques were induced by perivascular placement of constrictive collars around the carotid artery, intraperitoneal injection of lipopolysaccharide and stress stimulation. Mice were randomly divided into RNA interference (Ad-MCP-1i) group receiving local treatment of rAd5-MCP-1-shRNA suspension, Ad-EGFP group receiving treatment of rAd5-mediated negative shRNA and mock group receiving treatment of saline. Two weeks after treatment, plaque disruption rates were significantly lower in the Ad-MCP-1i group than in the Ad-EGFP group (13.3% vs. 60.0%, P = 0.01), and local MCP-1 expression was significantly inhibited in the Ad-MCP-1i group confirmed by immunostaining, qRT-PCR and western blot (P<0.001). Compared with the Ad-EGFP group, carotid plaques in the Ad-MCP-1i group showed increased levels of collagen and smooth muscle cells, and decreased levels of lipid and macrophages. The expression of inflammatory cytokines and activities of matrix metalloproteinases (MMPs) were lower in the Ad-MCP-1i group than in the Ad-EGFP group. In conclusion, site-specific delivery of adenoviral-mediated shRNA targeting mouse MCP-1 downregulated MCP-1 expression, turned a vulnerable plaque into a more stable plaque phenotype and prevented plaque disruption. A marked suppression of the local inflammatory cytokine expression may be the central mechanism involved.
Insights
Local RNA interference targeting monocyte chemoattractant protein-1 (MCP-1) stabilized atherosclerotic plaques in mice. This approach reduced plaque disruption and inflammation, offering a potential therapeutic strategy for atherosclerosis.
Area of Science:
- Cardiovascular Research
- Molecular Biology
- Gene Therapy
Background:
- Monocyte chemoattractant protein-1 (MCP-1), a CC chemokine (CCL2), plays a critical role in atherosclerosis development.
- MCP-1 is recognized as a significant therapeutic target for managing atherosclerosis.
Purpose of the Study:
- To investigate the efficacy of local RNA interference (RNAi) of MCP-1 in enhancing plaque stability.
- To assess the prevention of plaque disruption using adenovirus-mediated small hairpin RNA (shRNA) targeting MCP-1 in ApoE-/- mice.
Main Methods:
- Adenovirus-mediated shRNA targeting mouse MCP-1 (rAd5-MCP-1-shRNA) was designed and delivered site-specifically.
- Atherosclerotic plaques were induced in ApoE-/- mice fed a high-fat diet using constrictive collars, LPS, and stress.
- Mice were treated with rAd5-MCP-1-shRNA (Ad-MCP-1i), a negative control shRNA (Ad-EGFP), or saline (mock).
Main Results:
- Local MCP-1 expression was significantly inhibited in the Ad-MCP-1i group compared to controls (P<0.001).
- Plaque disruption rates were significantly lower in the Ad-MCP-1i group (13.3%) versus the Ad-EGFP group (60.0%, P = 0.01).
- Carotid plaques in the Ad-MCP-1i group exhibited increased collagen and smooth muscle cells, with decreased lipid and macrophage content, alongside reduced inflammatory cytokines and MMP activity.
Conclusions:
- Site-specific delivery of adenoviral-mediated shRNA targeting MCP-1 effectively downregulates MCP-1 expression.
- This intervention transforms vulnerable plaques into a more stable phenotype, preventing plaque disruption.
- Suppression of local inflammatory cytokine expression appears to be the primary mechanism underlying these effects.
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