Canine IL4-10 fusion protein provides disease modifying activity in a canine model of OA; an exploratory study

Abstract

Objective An ideal disease modifying osteoarthritis drug (DMOAD) has chondroprotective, anti-inflammatory, and analgesic effects. This study describes the production and characterization of a canine IL4-10 fusion protein (IL4-10 FP) and evaluates its in vivo DMOAD activity in a canine model of osteoarthritis (OA). Design The canine Groove model was used as an in vivo model of degenerative knee OA. Six weeks after OA induction dogs were intra-articularly injected weekly, for ten weeks, with either IL4-10 FP or phosphate buffered saline (PBS). In addition to the use of human IL4-10 FP, canine IL4-10 FP was developed and characterized in vitro , and tested in vivo . Force plate analysis (FPA) was performed to analyze joint loading as a proxy measure for pain. After ten weeks dogs were euthanized and cartilage and synovial tissue samples were analyzed by histochemistry (OARSI scores) and biochemistry (cartilage proteoglycan turnover). Results Repetitive intra-articular injections with human IL4-10 FP led to antibody formation, that blocked its functional activity. Therefore, a canine IL4-10 FP was developed, which completely inhibited LPS-induced TNFα production by canine blood cells, and increased proteoglycan synthesis of canine cartilage in vitro (p = 0.043). In vivo , canine IL4-10 FP restored the, by OA impaired, joint loading (p = 0.002) and increased cartilage proteoglycan content (p = 0.029). Conclusions This first study on the potential DMOAD activity upon prolonged repeated treatment with IL4-10 FP demonstrates that a species-specific variant has anti-inflammatory and chondroprotective effects in vitro and chondroprotective and analgesic effects in vivo . These data warrant further research on the DMOAD potential of the IL4-10 FP.

Experimental Therapeutics for the Treatment of Osteoarthritis

“…28,[100][101][102] Hence, an agonistic IL-4/IL-10 fusion antibody seems to be promising for OA therapy. 103 This therapeutical approach based on human IL-10 reduced pain in the dogs, as reported by the veterinarians and dog owners observations, without any detectable adverse effects. These results might provide a starting point for clinical trials to confirm it in future as effective.…”

IL4-10 Fusion Protein Shows DMOAD Activity in a Rat Osteoarthritis Model

“…Pain was assessed in a limited number of rats (6 per group), nevertheless, the results are in line with previous results, found in an in vivo canine model of OA. 19 In vitro, the activity of rIL4-10 FP is 10-fold lower compared to the activity of solely IL10 (Fig. 2D).…”

“…Production and characterization of rIL4-10 FP are largely done using the same procedures as published previously by this group for cIL4-10 FP. 19 transfection and Cell Culture. rIL4-10 FP was produced by transient transfection of HEK293F cells with a pcDNA3.1neo expression vector (Invitrogen, Carlsbad, CA, USA) containing a dual CMV (cytomegalovirus) promoter.…”

“…18 The chondroprotective and analgesic effects of IL4-10 FP were previously confirmed with intra-articular injections of canine IL4-10 FP in the canine OA Groove model. 19 To test the IL4-10 FP in a rodent model of OA, the rat Groove model was used. 20 This model of mechanically induced cartilage damage is mild and allows for tissue repair as there is no permanent trigger for joint damage (as, e.g., in instability or chemical induced models).…”

Novel Analgesics with Peripheral Targets

“…IL-4 receptors are found in both mouse and human sensory neurons and elicit functional responses [432]. A new concept is a fusion protein of two anti-inflammatory interleukins; the IL4-10 fusion protein has shown an analgesic effect in vivo in a model of osteoarthritis [433,434]. Related to IL-4 is IL-13, which is secreted by immune cells and changes the phenotype of local macrophages towards an anti-inflammatory phenotype.…”