Investigating the impact of interstitial fluid flow on cancer biology

Pyne, Emily Seton

Abstract

Despite major advances in our understanding of cancer progression mechanisms, cancer remains a leading cause of death in the UK and globally. Solid tumours are complex, and heterogeneous, existing in a dynamic microenvironment subjected to a variety of biomechanical cues. Conventional cancer research methods have long relied on two-dimensional (2D) static cultures which neglect the dynamic, three-dimensional (3D) nature of the tumour microenvironment (TME), especially the role of interstitial fluid (ISF) and the impact of interstitial fluid flow (IFF). In order to address this, we have developed and optimised a spheroid-on-chip microfluidic platform which allows 3D cancer spheroids to be integrated into extracellular matrices (ECM)-like hydrogels and exposed to continuous perfusion, mimicking IFF in the TME. Our platform enables clear imaging of spheroids, analysis of effluent media, and harvesting of spheroids for further analyses. Where many studies have focused on perfusion as a tool mainly for high-throughput methods, we aim to identify how perfusion, mimicking IFF in the TME, alters the biology of cancer spheroids.
Spheroids exposed IFF-like or static conditions were harvested, and gene and protein expression analysis was performed. These data indicated that gene expression was altered by flow when compared to static conditions, but clear functional patterns were not identifiable. Subsequently, a large-scale transcriptomic analysis by RNA-sequencing was used to conduct an unbiased analysis, revealing robust gene expression changes in genome preservation pathways and leading to the identification of a potential biomarker of IFF in cancer.
Our spheroid-on-chip flow platform has revealed that exposure to IFF-like conditions in a 3D environment significantly altered the transcriptome of spheroids after just 24 hours, leading to changes in the expression of key factors involved in various aspects of tumour progression. These results will direct our future work on further elucidating the role of ISF flow in cancer biology and spread.

Citation

Pyne, E. S. (2022). Investigating the impact of interstitial fluid flow on cancer biology. (Thesis). University of Hull. Retrieved from https://hull-repository.worktribe.com/output/4251436

• Thesis Type: Thesis
• Deposit Date: Mar 27, 2023
• Publicly Available Date: Jan 31, 2024
• Keywords: Biomedical sciences
• Public URL: https://hull-repository.worktribe.com/output/4251436
• Additional Information: Centre for Biomedicine, The University of Hull
• Award Date: 2022-11