Evaluation of Matrix Metalloproteinase Inhibition by Peptide Microarray-Based Fluorescence Assay on Polymer Brush Substrate and in Vivo Assessment

• Published in: ACS applied materials & interfaces 2017-12, Vol.9 (50), p.44241-44250
• Main Authors: Lei, Zhen, Chen, Hongda, Zhang, Hua, Wang, Yaoqi, Meng, Xianying, Wang, Zhenxin
• Format: Article
• Language: English
• Subjects: Animals; Cell Line, Tumor; Fluorescent Dyes; Humans; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Mice; Peptides; Polymers; Protein Array Analysis
• ISSN: 1944-8244; 1944-8252
• DOI: 10.1021/acsami.7b15445

Matrix metalloproteinases (MMPs) are important biomarkers and potential therapeutic targets of tumor. In this report, a peptide microarray-based fluorescence assay is developed for MMPs inhibitors evaluation through immobilization of biotin-modified peptides on the poly­(glycidyl methacrylate-co-2-hydroxyethyl methacrylate) (P­(GMA-HEMA)) brush-modified glass slides. After biotin is recognized with cyanine 3 (Cy3)-modified avidin (Cy3-avidin), the microarrays can produce strong fluorescence signal. The biotin moieties detach from microarray, when the biotin-modified peptide substrates are specially cleaved by a MMP, resulting in decreased fluorescence intensity of the microarray. The decreasing level of fluorescence intensity is correlated with the MMP inhibition. Nine known MMP inhibitors against MMP-2 and MMP-9 are evaluated by the assay, and the quantitative determination of inhibitory potencies (half maximal inhibitory concentration) are obtained, which are comparable with the literatures. Two biocompatible fluorogenic peptides containing MMP-specific recognition sequences and FAM/Dabcyl fluorophore-quencher pair are designed as activatable reporter probes for sensing MMP-2 and MMP-9 activities in cell and in vivo. The peptide microarray-based results are well verified by the cell inhibition assay and in vitro fluorescence imaging, and further confirmed by the in vivo imaging of HT-1080 tumor-bearing mice.