Abstract

Mechanobiological response plays an important role in modulating cell identity. A multitude of studies have shown that different biophysical aspects of the microenvironments have profound impact on the cell both in physiological as well as in pathological conditions like in cancer, where it contributes in determining the prognosis of the disease. Recent publications have shown that mechanobiological stimulation may affect chromatin structure and gene regulation but the mechanism behind this response is still largely unknown. Pancreatic ductal adenocarcinoma (PDAC) is extremely aggressive and its bad prognosis strongly correlates with the high stiffness of the extracellular matrix. However, the lack of studies addressing the contribution of other biophysical properties to PDAC progression represents a big gap in the field. Here we present preliminary data in which we characterized the response of PDAC cells that were grown on engineered nanostructured surfaces. The results show that the nanotopography affects cell mechanics and in turn induces morphological changes as well as their migratory parameters. Strikingly, nanotopographical features also trigger functional response in PDAC cells by changing their sensitivity to specific chemotherapy drugs. Determining the interplay between the physical environment with the single-cell transcriptional profiling is key for the understanding of how mechanotransduction regulates gene expression in cancer progression. FluidFM live-seq is a novel method that allows gene expression profiling of living single cells. Using FluidFM Nano-Syringes, biopsies can be directly retrieved from a cell and then analyzed with a highly-sensitive, low-input RNA-seq protocol. The gene expression profiles obtained from Live-seq are faithful representations of single cell transcriptomes, thus enabling to take transcriptome snapshots of a cell without lysing it. By Using FluidFM we aim to reveal key insights about the dynamic response of PDAC cells to the different substrates, but also emphasize the phenotypic traits based on their morphology that may be too infrequent or transitory to be captured by NGS approaches.