title: A short DNA aptamer that recognizes TNFα and blocks its activity in vitro.
abstract: Tumor necrosis factor-alpha (TNFα) is a pivotal component of the cytokine network linked to inflammatory diseases. Protein-based, TNFα inhibitors have proven to be clinically valuable. Here, we report the identification of short, single-stranded DNA aptamers that bind specifically to human TNFα. One such 25-base long aptamer, termed VR11, was shown to inhibit TNFα signaling as measured using NF-κB luciferase reporter assays. This aptamer bound specifically to TNFα with a dissociation constant of 7.0 ± 2.1 nM as measured by surface plasmon resonance (SPR) and showed no binding to TNFβ. Aptamer VR11 was also able to prevent TNFα-induced apoptosis as well as reduce nitric oxide (NO) production in cultured cells for up to 24 h. As well, VR11, which contains a GC rich region, did not raise an immune response when injected intraperitoneally into C57BL/6 mice when compared to a CpG oligodeoxynucleotide (ODN) control, a known TLR9 ligand. These studies suggest that VR11 may represent a simpler, synthetic scaffold than antibodies or protein domains upon which to derive nonimmunogenic oligonucleotide-based inhibitors of TNFα.
authors.affiliation: Department of Pharmaceutical Sciences, Leslie Dan Faculty of Pharmacy, University of Toronto, 144 College Street, Toronto, Ontario M5S 3M2, Canada.
authors.forename: Erik W
authors.initials: EW
authors.lastname: Orava
authors.forename: Nick
authors.initials: N
authors.lastname: Jarvik
authors.forename: Yuen Lai
authors.initials: YL
authors.lastname: Shek
authors.forename: Sachdev S
authors.initials: SS
authors.lastname: Sidhu
authors.forename: Jean
authors.initials: J
authors.lastname: Gariépy
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide / Q000494:pharmacology*; D001483:Base Sequence; D002460:Cell Line; D002942:Circular Dichroism; D005347:Fibroblasts; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D020349:Surface Plasmon Resonance; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors*
chemical_list: D052157:Aptamers, Nucleotide; D014409:Tumor Necrosis Factor-alpha