IL-17RA aptamer-mediated repression of IL-6 inhibits synovium inflammation in a murine model of osteoarthritis

Abstract

Generate DNA aptamers to inhibit IL-17RA-mediated synovial inflammation in an experimental mouse model of osteoarthritis (OA). A novel cell-SELEX method was applied to obtain DNA aptamers specific for IL-17RA. A single-stranded (ss) DNA library with four(30) probes was synthesised. By incubating this library with NIH3T3 cells, we collected DNA ligands that could bind the cell surface. The collected ligands were incubated with IL-17RA-deficient NIH3T3 cells, and unbound ssDNA was harvested from the supernatant for the next round of selection. After 12 cycles, specific aptamers against IL-17RA were generated. For animal experiments, a meniscectomy was performed on Balb/C mice to generate an animal model of OA. Mice received weekly intra-articular (i.a.) injections of aptamers or control treatments for 6 weeks. Synovial membranes were evaluated by histomorphology and the mRNAs of critical inflammatory cytokines were measured by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). An aptamer termed RA10-6 was obtained that could efficiently block IL-17 binding to IL-17RA in a dose-dependent manner in vitro. Histological examination and quantitative RT-PCR results showed that OA mice that injected with RA10-6, especially in combination with celecoxib demonstrated inhibition of synovial thickening and reduction in IL-6 levels in the synovial tissue. Our results suggest that RA10-6 can inhibit synovial inflammation by blocking IL-17/IL-17RA-mediated IL-6 expression. RA10-6 acted synergistically with celecoxib to inhibit IL-6 expression in synovial tissues. Thus, aptamers targeting IL-17RA might serve as potent adjunctive agents for the early treatment of OA.