Protective effect of lentivirus-mediated siRNA targeting ADAMTS-5 on cartilage degradation in a rat model of osteoarthritis

Abstract

The etiology of osteoarthritis (OA) is complex and multifaceted. Osteoarthritis is a chronic and progressive disease of the joints that is characterized by the degradation of articular cartilage. A disintegrin and metalloproteinase with thrombospondin motifs-5 (ADAMTS-5) is the major aggrecanase in cartilage. The aim of this study was to evaluate the effect of ADAMTS-5 knockdown on cartilage degradation. Rat articular chondrocytes were transfected with lentivirus‑mediated ADAMTS-5 small interfering RNA (siRNA) or with empty vector control plasmid DNA (as the control). The suppression efficiency was measured using real-time polymerase chain reaction (RT-PCR) and western blot analysis. We then selected the most effective siRNA (siRNA1) and constructed the lentivirus-mediated siRNA targeting ADAMTS-5 for stable transfection. An animal model of OA was created using male Sprague-Dawley rats. OA was induced by performing anterior cruciate ligament transection (ACL-T) and partial medial meniscectomy (PM). The animals (n=80, weight 250‑300 g) received an intra‑articular injection of the empty vector control plasmid DNA or lentivirus‑mediated ADAMTS-5 siRNA1 (20 µl, 1x108 TU/ml). The progression of OA was analyzed using Osteoarthritis Research Society International (OARSI) scores. Compared with the control, ADAMTS-5 gene expression was decreased by approximately 80% by siRNA1 in a monolayer culture of chondrocytes. The intra‑articular injection of lentivirus‑mediated ADAMTS-5 siRNA1 in vivo resulted in the downregulation of ADAMTS-5 protein expression and improved OARSI scores (p<0.05). A single injection of lentivirus‑mediated ADAMTS‑5 siRNA prevented the degradation of articular cartilage. This method may provide a novel therapeutic strategy for the treatment of human OA.

Pterostilbene inhibits inflammation and ROS production in chondrocytes by activating Nrf2 pathway

“…The osteoarthritis Sprague-Dawley (SD) rat model was established as previously described [38]. Briefly, 18 male 200–300 g SD (2 months old) rats in the conventional housing were anesthetized by the10% (w/v) chloral hydrate (0.2 mL/100 g) by intraperitoneal injection and randomly divided into 3 groups including normal, anterior cruciate ligament (ACL) transection and partial medial meniscectomy (ACLX), and ACLX+PTE groups.…”

rAAV‐mediated combined gene transfer and overexpression of TGF‐β and SOX9 remodels human osteoarthritic articular cartilage

“…In this regard, gene therapy offers powerful tools to counteract the progressive degradation of the cartilage in OA as it permits a sustained production of therapeutic factors that otherwise display short pharmacological half‐lives. Therapeutic gene transfer has been successfully tested to either protect the OA joint from cartilage breakdown by applying sequences coding for protective or inhibitory factors (IL‐1 receptor antagonist—IL‐1Ra, soluble IL‐1, and TNF receptors—sIL‐1R and sTNFR, siRNAs/shRNAs against IL‐1 or ADAMTS‐5, NF‐κB inhibitors, kallistatin—KBP, thrombospontin‐1—TSP‐1, Dickkopf‐1—DKK‐1, pro‐opiomelanocortin—POMC, the transcriptional repressor Bagpipe homeobox homolog 1/NK3 homeobox 2—BAPX1/NKX3.2, miRNAs) or enhance anabolic processes by gene transfer of proteoglycan 4, glucuronosyltransferase‐I, the insulin‐like growth factor I (IGF‐I), fibroblast growth factor 2 (FGF‐2), and the bone morphogenetic proteins 4 and 7 (BMP‐4, ‐7) . However, none of such attempts thus far were capable of re‐establishing a natural balance in OA cartilage to restore the full integrity of the damaged tissue, and multiple treatments instead of single options might be necessary to reproduce an original cartilage architecture in OA as already evaluated using dual gene transfer of IL‐1Ra/IGF‐I or/FGF‐2 or of FGF‐2/SOX9 .…”

Small non-coding RNAs-based bone regulation and targeting therapeutic strategies

“…It was corroborated that overexpression of ADAMTS-5 mediated by IL-1b induced aggravated degradation of cartilage matrix in OA and significantly inhibited miR-30a expression levels (Ji et al, 2016a). siRNA-mediated blockage of ADAMTS-5 demonstrated protective effects on cartilage degradation (Chu et al, 2013). Moreover, intra-articular delivery of hypoxia-inducible factor-2a (HIF-2a) siRNA by chondrocyte-homing nanoparticles resulted in considerably decreased expression of some catabolic proteins involved in the progression of OA, such as MMP-13, VEGF, and ADAMTS-5, thus providing better prevention of cartilage degeneration in OA patients (Pi et al, 2015).…”