microRNA-181a-5p antisense oligonucleotides attenuate osteoarthritis in facet and knee joints

Abstract

ObjectivesWe recently identified microRNA-181a-5p (miR-181a-5p) as a critical mediator involved in the destruction of lumbar facet joint (FJ) cartilage. In this study, we tested if locked nucleic acid (LNA) miR-181a-5p antisense oligonucleotides (ASO) could be used as a therapeutic to limit articular cartilage degeneration.MethodsWe used a variety of experimental models consisting of both human samples and animal models of FJ and knee osteoarthritis (OA) to test the effects of LNA-miR-181a-5p ASO on articular cartilage degeneration. Histopathological analysis including immunohistochemistry and in situ hybridisation were used to detect key OA catabolic markers and microRNA, respectively. Apoptotic/cell death markers were evaluated by flow cytometry. qPCR and immunoblotting were applied to quantify gene and protein expression.ResultsmiR-181a-5p expression was increased in human FJ OA and knee OA cartilage as well as injury-induced FJ OA (rat) and trauma-induced knee OA (mouse) cartilage compared with control cartilage, correlating with classical OA catabolic markers in human, rat and mouse cartilage. We demonstrated that LNA-miR-181a-5p ASO in rat and mouse chondrocytes reduced the expression of cartilage catabolic and chondrocyte apoptotic/cell death markers in vitro. Treatment of OA-induced rat FJ or mouse knee joints with intra-articular injections of in vivo grade LNA-miR-181a-5p ASO attenuated cartilage destruction, and the expression of catabolic, hypertrophic, apoptotic/cell death and type II collagen breakdown markers. Finally, treatment of LNA-miR-181a-5p ASO in cultures of human knee OA chondrocytes (in vitro) and cartilage explants (ex vivo) further demonstrated its cartilage protective effects.ConclusionsOur data demonstrate, for the first time, that LNA-miR-181a-5p ASO exhibit cartilage-protective effects in FJ and knee OA.

Osteoarthritis year in review 2019: genetics, genomics and epigenetics

“…A second study focused on miR-181a-5p, which is increased in degenerated human facet joints and OA knee cartilage, and during mouse OA development 64 . Injection of antisense oligonucleotides, which attenuate miR-181a-5p activity, reduced cartilage damage in knee joints with concomitant reduction in catabolic gene expression and markers of cartilage damage.…”

Response to: ‘Regarding microRNA-181a-5p antisense oligonucleotides attenuate osteoarthritis in facet and knee joints’ by Liebling

“…We wish to thank Dr Liebling for the relevant comments1 regarding our recent article in Annals of the Rheumatic Diseases entitled ‘MicroRNA-181a-5p antisense oligonucleotides attenuate osteoarthritis in facet and knee joints’ 2. We are delighted that our article has gathered such interest in the scientific community and are happy to provide additional comments.…”

“…This effect is not observed in mouse or rat cells (online supplementary figures 3 and 4). Thus, this human chondrocyte response could be a result of species-selective ‘off-target effects’ of the in vitro grade LNA-miR-181a-5p ASO, which we concede as a limitation in the discussion section of our published work 2. However, LNA-miR-181a-5p ASO is effective at reducing chondrocyte cell death in vitro and in vivo under disease-mimicking conditions (with either IL-1β-treatment or surgical induction).…”

“…We are delighted that our article has gathered such interest in the scientific community and are happy to provide additional comments. In the aforementioned publication, we determined the potential of locked nucleic acid, antisense oligonucleotides against microRNA-181a-5p (LNA-miR-181a-5p ASO) as a therapy for the treatment of osteoarthritis (OA) 2. The chemical modification of LNA is one of the most advanced ASO delivery systems.…”

Regarding micro-RNA-181a-5p antisense oligonucleotides attenuate osteoarthritis in facet and knee joints

“…I read with considerable interest the paper by Nakamura and colleagues dealing with the potential therapeutic use of a locked nucleic acid, antisense oligonucleotide (LNA ASO) against a micro-RNA in osteoarthritis (OA) 1. I was particularly impressed with the Osteoarthritis Research Society International and cellularity scores as well as the data on degenerative markers and chondrocyte apoptosis/death from the in vivo animal models.…”