EGFP-EGF1-conjugated poly (lactic-co-glycolic acid) nanoparticles as a carrier for the delivery of CCR2− shRNA to atherosclerotic macrophage in vitro

• Published in: Scientific reports 2020-11, Vol.10 (1), p.19636, Article 19636
• Main Authors: Wu, Zhilin, Chen, Chen, Luo, Jiajia, Davis, Jacques R. J., Zhang, Bo, Tang, Liang, Shi, Wei, Liao, Danying
• Format: Article
• Language: English
• Subjects: 631/61/54/152; 692/699; 692/699/75/593/2100; Animals; Arteriosclerosis; Atherosclerosis; Atherosclerosis - drug therapy; Atherosclerosis - metabolism; Biocompatible Materials - chemistry; CC chemokine receptors; CCR2 protein; Cell Line; Coumarin; Cytotoxicity; Drug Carriers - administration & dosage; Drug delivery; Drug Delivery Systems; Feasibility studies; Flow cytometry; Fluorescence microscopy; Glycolic acid; Green Fluorescent Proteins - chemistry; Humanities and Social Sciences; Inflammation; Macrophages; Macrophages - drug effects; Macrophages - metabolism; Mice; Monocyte chemoattractant protein 1; Monocytes; mRNA; multidisciplinary; Nanoparticles; Nanoparticles - administration & dosage; Nanoparticles - chemistry; Peptide Elongation Factor G - chemistry; Polylactic Acid-Polyglycolic Acid Copolymer - chemistry; Polylactide-co-glycolide; Receptors, CCR2 - antagonists & inhibitors; Receptors, CCR2 - genetics; Recombinant Fusion Proteins - chemistry; RNA, Small Interfering - administration & dosage; RNA, Small Interfering - chemistry; RNA, Small Interfering - pharmacology; Science; Science (multidisciplinary); siRNA; Tissue factor; Transfection
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-020-76416-4

Reducing macrophage recruitment by silencing chemokine (C–C motif) receptor 2 (CCR2) expression is a promising therapeutic approach against atherosclerosis. However the transfection of macrophages with siRNA is often technically challenging. EGFP-EGF1-conjugated poly (lactic-co-glycolic acid) (PLGA) nanoparticles (ENPs) have a specific affinity to tissue factor (TF). In this study, the feasibility of ENPs as a carrier for target delivery of CCR2-shRNA to atherosclerotic cellular models of macrophages was investigated. Coumarin-6 loaded ENPs were synthesized using a double-emulsion method. Fluorescence microscopy and flow cytometry assay were taken to examine the uptake of Coumarin-6 loaded ENPs in the cellular model. Then a sequence of shRNA specific to CCR2 mRNA was constructed and encapsulated into ENPs. Target delivery of CCR2-shRNA to atherosclerotic cellular models of macrophages in vitro were evaluated. Results showed more uptake of ENPs by the cellular model than common PLGA nanoparticles. CCR2-shRNA loaded ENPs effectively silenced CCR2 gene in the atherosclerotic macrophages and exhibited a favorable cytotoxic profile to cultured cells. With their low cytotoxicity and efficient drug delivery, ENP could be a useful carrier for target delivery of CCR2-shRNA to inflammatory monocytes/macrophages for the therapy against atherosclerosis.