Dual-active hyaluronidase D016 reduces established atherosclerotic lesion area in LDLR⁻/⁻ mice

Pathological accumulation of extracellular-matrix components, including hyaluronan and sulfated glycosaminoglycans (sGAGs), has been suggested to sustain advanced atherosclerotic plaques by altering tissue architecture and transport. D016 is a recombinant dual-active hyaluronidase designed to cleave both substrates, aiming to modulate matrix-associated components in the plaque microenvironment rather than lipid metabolism. Male LDLR−/− mice with established diet-induced atherosclerosis received intravenous vehicle or D016 once daily for 10 days (7,500 or 750,000 IU kg − 1), followed by a 19-day post-treatment interval. Aortic-root lesion area was quantified histologically on day 29. High-dose D016 was associated with an ~ 19% reduction in aortic-root plaque area versus vehicle (P < 0.05, one-way ANOVA with Dunnett’s test; n = 12 per group); the low dose showed no detectable effect. Protease-inhibitor co-treatment did not further reduce lesion area. This study did not quantify plaque ECM composition and therefore does not establish demonstrable ECM remodelling. An effect on lesion progression cannot be excluded under continued Western diet. Clinical condition, body weight, and hepatic and renal serum parameters were similar between groups. These results support further mechanistic evaluation of enzymatic modulation of matrix-associated pathways.