BPS2026 – The role of matrix viscoelasticity on mediating collective invasion in liver cancer

Hepatocellular carcinoma (HCC) is the most common liver cancer and has a poor prognosis due to its invasive nature. The current paradigm is that increased stiffness during cirrhosis is a critical driver of HCC progression, with cells sensing the collagen-induced stiffness, and becoming more invasive. While the bulk stiffness increases during cirrhosis, the architecture of the cirrhotic liver is very heterogenous. Here, we examine how the heterogenous matrix mechanics mediate the process of collective invasion by the HCC cells. One component of the heterogenous matrix is the regenerative areas which is where HCC arise from hepatocytes in the dysplastic nodules. We have found that regenerative areas in patients with metabolic dysfunction associated steatohepatitis (MASH)-related cirrhosis and in animal models are more viscoelastic than the healthy liver (faster stress relaxation). We use collagen-alginate interpenetrating networks of varying stiffness and stress relaxation for 3D cell culture studies to probe how changes in matrix mechanics impact HCC collective invasion in various cell types. We found that differentiated, epithelial HCC (Huh7) and de-differentiated HCC (SNU449) cells invade collectively in faster stress relaxing CAiPNs compared to their slower stress relaxing counterparts at the same stiffness (∼2.5 kPa). The Huh7 spheroids invade in a traditional collective manner—with a multicellular protrusion whereas the SNU449 spheroids invade in a cell streaming manner. While MMP inhibition has minimal effect on invasion, inhibition of integrin β1 potently diminishes invasion, suggesting the role integrin β1-mediated mechanotransduction and force transmission. Our ongoing work is examining the role of cytoskeletal proteins and metabolism (glycolysis vs. oxidative phosphorylation) in mediating the impact of viscoelasticity on invasion. Together, these results point to changes in matrix viscoelasticity playing a key role in mediating early collective HCC invasion.