Validation of the exosomal protein SERPINA11 as a potential atherosclerosis marker via bioprinted scaffold.

Abstract

Existing animal and human cell models have limitations in terms of heterogeneous differences or difficulties in sufficiently reproducing arterial structures and complex cell-cell interactions. The discovery of exosome-derived biomarkers using a three-dimensional (3D) bioprinted atherosclerosis model provides a noninvasive and stable detection method and is expected to contribute to the development of early diagnosis and personalized treatment. To contribute to the discovery of exosome-derived biomarkers related to the early diagnosis and prognosis of cardiovascular diseases using a 3D bioprinted atherosclerosis model, we reproduced an arterial environment using 3D bioprinting composed of a biocompatible extracellular matrix (bioink) and various human cells<i>in vitro</i>. The 3D bioprinted atherosclerosis model composed of inflammatory macrophages, coronary artery smooth muscle cells, coronary artery endothelial cells, and collagen methacryloyl (ColMA) hydrogel was treated with low-density lipoproteins to induce atherosclerosis, and the atherosclerosis model was classified into Baseline, early atherosclerosis (EA; Early Athero), and late atherosclerosis (LA; Late Athero) groups. The secreted exosomes were isolated according to the time period, and a characterization analysis was conducted to confirm the purity of the isolated exosomes. We evaluated the isolated exosomes qualitatively and quantitatively. Isolated exosomes were analyzed using proteomics and microRNA (miRNA) sequencing to verify whether the bioprinted atherosclerosis model induced atherosclerosis, and a novel EA biomarker, SERPINA11, was discovered. In conclusion, we verified that the bioprinted atherosclerosis model induced atherosclerosis and that the novel biomarker set of exosomal miRNAs (hsa-miR-143-5p and hsa-miR-6879-5p) expressed in EA and proteins (SERPINA11, AHSG, and F2) might be clinically useful in early diagnosis and prognosis.