title: A two-step stimulus-response cell-SELEX method to generate a DNA aptamer to recognize inflamed human aortic endothelial cells as a potential in vivo molecular probe for atherosclerosis plaque detection.
abstract: Aptamers are single-stranded oligonucleotides that are capable of binding wide classes of targets with high affinity and specificity. Their unique three-dimensional structures present numerous possibilities for recognizing virtually any class of target molecules, making them a promising alternative to antibodies used as molecular probes in biomedical analysis and clinical diagnosis. In recent years, cell-systematic evolution of ligands by exponential enrichment (SELEX) has been used extensively to select aptamers for various cell targets. However, aptamers that have evolved from cell-SELEX to distinguish the "stimulus-response cell" have not previously been reported. Moreover, a number of cumbersome and time-consuming steps involved in conventional cell-SELEX reduce the efficiency and efficacy of the aptamer selection. Here, we report a "two-step" methodology of cell-SELEX that successfully selected DNA aptamers specifically against "inflamed" endothelial cells. This has been termed as stimulus-response cell-SELEX (SRC-SELEX). The SRC-SELEX enables the selection of aptamers to distinguish the cells activated by stimulus of healthy cells or cells isolated from diseased tissue. We report a promising aptamer, N55, selected by SRC-SELEX, which can bind specifically to inflamed endothelial cells both in cell culture and atherosclerotic plaque tissue. This aptamer probe was demonstrated as a potential molecular probe for magnetic resonance imaging to target inflamed endothelial cells and atherosclerotic plaque detection.
authors.affiliation: NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, Singapore, 119077, Singapore.
authors.forename: Kaili
authors.initials: K
authors.lastname: Ji
authors.forename: Wee Siang
authors.initials: WS
authors.lastname: Lim
authors.forename: Sam Fong Yau
authors.initials: SF
authors.lastname: Li
authors.forename: Kishore
authors.initials: K
authors.lastname: Bhakoo
mesh_terms: D001025:Aortitis / Q000378:metabolism* / Q000473:pathology*; D052157:Aptamers, Nucleotide / Q000138:chemical synthesis / Q000378:metabolism*; D002478:Cells, Cultured; D003324:Coronary Artery Disease / Q000175:diagnosis* / Q000378:metabolism*; D042783:Endothelial Cells / Q000378:metabolism* / Q000473:pathology; D005240:Feasibility Studies; D006801:Humans; D015336:Molecular Probe Techniques; D052156:SELEX Aptamer Technique
chemical_list: D052157:Aptamers, Nucleotide