Engineered protein nanoclusters reduce liver fibrosis and hepatocellular carcinoma in mice models.

Long-term liver injury leads to acute hepatitis and chronic liver diseases, including fibrosis and hepatocellular carcinoma (HCC), which frequently necessitate liver transplantation and are associated with poor median survival. Transforming growth factor-β (TGF-β) signalling pathway is a key driver of fibrogenesis and tumor progression. Therapeutic approaches targeting this pathway-via TGF-β receptor blockade or inhibition of its chaperone protein Hsp90- are under pre-clinical and clinical trials, and despite some promising results none of them has been adopted as a definitive therapy. Engineered protein scaffolds have emerged as an attractive alternative owing to their high engineerability and ease of production. In particular, consensus tetratricopeptide repeat (CTPR) proteins are strong candidates because of their modularity, robustness, design flexibility, and amenability to functionalization. Here, we investigate the protein-nanocluster hybrid formulation comprising C390 and nanocluster-stabilizing domain (C390-AuNC) as a Hsp90 inhibitory platform to assess its potential dual anti-fibrotic and anti-tumor activity in two chemically-induced murine liver disease models: acute liver fibrosis and chronic HCC. In the fibrosis model, C390-AuNC markedly suppress the expression of profibrotic markers and promote degradation of collagen fibers in the liver, indicating effective attenuation of fibrogenesis. In the HCC model, C390-AuNC inhibit Hsp90, leading to reduced expression of oncogenic proteins that drive cancer cell proliferation and metastasis, and consequently diminish tumor burden. Collectively, these findings support C390-AuNC as a promising next-generation biotherapeutic platform with low anticipated immunogenicity and high therapeutic potential.