Extracellular Matrix Viscoelasticity Promotes Liver Cancer Progression in Pre-Cirrhotic NASH

Type 2 diabetes mellitus (T2DM) is a major risk factor for hepatocellular carcinoma (HCC). Changes in extracellular matrix (ECM) mechanics contribute to cancer development, and increased stiffness is known to promote HCC progression in cirrhotic conditions. T2DM is characterized by an accumulation of advanced glycation end products (AGEs) in the ECM; however, how this affects HCC in non-cirrhotic conditions is unclear. Here, we find that in patients and animal models AGEs promote changes in collagen architecture and enhance ECM viscoelasticity, with greater viscous dissipation and faster stress relaxation, but not changes in stiffness. High AGEs and viscoelasticity combined with oncogenic b-catenin signaling promote HCC induction, while inhibiting AGEs production, reconstituting the clearance receptor AGER1, or breaking AGE-mediated collagen crosslinks reduce viscoelasticity and HCC growth. Matrix analysis and computational modeling demonstrate that lower interconnectivity of AGEs-bundled collagen matrix, marked by shorter fiber length and greater heterogeneity, enhance viscoelasticity. Mechanistically, animal studies and 3D cell cultures show that enhanced viscoelasticity promotes HCC cell proliferation and invasion through an integrin ß1–Tensin 1–YAP mechanotransduction pathway. These results reveal for the first time that AGEs-mediated structural changes enhance ECM viscoelasticity, and that viscoelasticity can drive cancer progression in vivo, independent of stiffness. Overall design: Ragefl/fl (WT) mice were subjected to chow, FFD, or HiAD for 14 weeks. A group of HiAD-fed mice was injected i.p. daily with PM. A cohort of RageHepKO mice was placed on HiAD for 14 weeks as published10. RNA was prepared from three mice per each group, and RNA sequencing was performed at Novogene (USA). Gencode gene annotations version M18 and the mouse reference genome major release GRCm38 was derived from https://www.gencodegenes.org/. Dropseq tools v1.1249 were used for mapping the raw sequencing data to the reference genome.