Plaque-targeted, proteolysis-resistant, activatable and MRI-visible nano-GLP-1 receptor agonist targets smooth muscle cell differentiation in atherosclerosis

• Published in: Theranostics 2022-01, Vol.12 (6), p.2741-2757
• Main Authors: Maiseyeu, Andrei, Di, Lin, Ravodina, Anastasia, Barajas-Espinosa, Alma, Sakamoto, Atsushi, Chaplin, Alice, Zhong, Jixin, Gao, Huiyun, Mignery, Matthew, Narula, Navneet, Finn, Aloke V, Rajagopalan, Sanjay
• Format: Article
• Language: English
• Subjects: Animals; Atherosclerosis; Atherosclerosis - diagnostic imaging; Atherosclerosis - drug therapy; Cardiovascular system; Cell Differentiation; Diet; Euthanasia; Experiments; GLP-1 receptor agonists; Glucagon-Like Peptide-1 Receptor - agonists; Hybridization; Inflammation - metabolism; Laboratories; Liraglutide - pharmacology; Magnetic Resonance Imaging; Membranes; Mice; Mice, Inbred C57BL; Mice, Knockout, ApoE; Muscle, Smooth - metabolism; Nanoparticles; Plaque, Atherosclerotic - diagnostic imaging; Plaque, Atherosclerotic - drug therapy; Proteolysis; Research Paper; RNA, Messenger - metabolism; Smooth muscle
• ISSN: 1838-7640; 1838-7640
• DOI: 10.7150/thno.66456

Glucagon-like peptide-1 receptor (GLP-1R) agonists are powerful glycemia-lowering agents, which have systematically been shown to lower cardiovascular events and mortality. These beneficial effects were difficult to pinpoint within atherosclerotic plaque due to lack of particular specificity of such agonists to the vascular cells and an inadequate understanding of the GLP-1R expression in atherosclerosis. Here, we hypothesized that the direct engagement of the GLP-1R in atherosclerosis by targeted agonists will alleviate vascular inflammation and plaque burden, even at a very low dose. The expression of GLP-1 receptor (GLP-1R, mRNA) in human lesions with pathologic intimal thickening, Apoe mouse atheroma and cultured immune/non-immune cells was investigated using genetic lineage tracing, Southern blotting and validated antisera against human GLP-1R. Protease-resistant and "activatable" nanoparticles (NPs) carrying GLP-1R agonist liraglutide (GlpNP) were engineered and synthesized. Inclusion of gadolinium chelates into GlpNP allowed for imaging by MRI. Atherosclerotic Apoe mice were treated intravenously with a single dose (30 µg/kg of liraglutide) or chronically (1 µg/kg, 6 weeks, 2x/week) with GlpNP, liraglutide or control NPs, followed by assessment of metabolic parameters, atheroma burden, inflammation and vascular function. Humal plaque specimens expressed high levels of GLP-1R within the locus of de-differentiated smooth muscle cells that also expressed myeloid marker CD68. However, innate immune cells under a variety of conditions expressed very low levels of , as seen in lineage tracing and Southern blotting experiments examining full-length open reading frame mRNA transcripts. Importantly, de-differentiated vascular smooth muscle cells demonstrated significant expression levels, suggesting that these could represent the cells with predominant positivity in atherosclerosis. GlpNP resisted proteolysis and demonstrated biological activity including glycemia lowering at 30 µg/kg and cholesterol efflux. Activatable properties of GlpNP were confirmed by imaging cytometry and using whole organ imaging. GlpNP targeted CD11b /CD11c cells in circulation and smooth muscle cells in aortic plaque in Apoe mice when assessed by MRI and fluorescence imaging. At a very low dose of 1 µg/kg, previously known to have little effect on glycemia and weight loss, GlpNP delivered i.v. for six weeks reduced triglyceride-rich lipoproteins in plasma, plaque burden and plaque