Stromal cells regulate mechanics of tumour spheroid

The remarkable contractility and force generation ability exhibited by cancer cells empower them to overcome the resistance and steric hindrance presented by a three-dimensional, interconnected matrix. Cancer cells disseminate by actively remodelling and deforming their extracellular matrix (ECM). The process of tumour growth and its ECM remodelling have been extensively studied, but the effect of the cellular tumour microenvironment (TME) has been ignored in most studies that investigated tumour-cell-mediated ECM deformations and realignment. This study reports the integration of stromal cells in spheroid contractility assays that impacts the ECM remodelling and invasion abilities of cancer spheroids. To investigate this, we developed a novel multilayer in vitro assay that incorporates stromal cells and quantifies the contractile deformations that tumour spheroids exert on the ECM. We observed a negative correlation between the spheroid invasion potential and the levels of collagen deformation. The presence of stromal cells significantly increased cancer cell invasiveness and altered the cancer cells' ability to deform and realign collagen gel, due to upregulation of proinflammatory cytokines. Interestingly, this was observed consistently in both metastatic and non-metastatic cancer cells. Our findings contribute to a better understanding of the vital role played by the cellular TME in regulating the invasive outgrowth of cancer cells and underscore the potential of utilising matrix deformation measurements as a biophysical marker for evaluating invasiveness and informing targeted therapeutic opportunities.Acknowledgements: We would like to thank all members of the EM and FC laboratory as well as L Ruban (UCL) for their help and support. AA, EM, and FC are grateful for the financial support by the Cancer Research UK Multidisciplinary Award (C57744/A22057). EM acknowledge financial support by Welcome Trust-MIT Fellowship (WT103883), Leverhulme Trust Research Project Grant (RPG-2018-443), BBSRC (BB/V001418/1). FC was funded by MCIN/AEI/10.13039/501100011033 (RYC-2016- 20352, RTI2018-096778-A-I00, PID2021-128107OB-I00); AECC (LAB-AE19044CALV, PRYCO211372RODR); BBVA Leonardo Awards (IN[19] _BBM_BAS_0076), and the ERC (Consolidator Grant 101045756)