Abstract 5034: Time-lapse analysis of drug response and invasive capacity using patient-derived CRC microfluidic 3D cancer models under oxygen-controlled conditions
Authors: Sabino Dora , Fixe Isabelle , Foucher Alexandra , Mennesson Eric , Normand Nadia 
Published: August 2018

The development of relevant, reliable, human-derived, and cost-effective cellular models is a present need to bring precision medicine to fruition and to accelerate drug discovery. 3D cultures show robust genomic stability and tissue-specific gene expression with high fidelity to their tissue of origin, making them ideal for preserving healthy and cancer tissue molecular and morphological traits. Our objective is to expand the number of relevant human cancer models available and implement methods that allow cellular response quantification of 3D cultures. We sought to establish and implement 3D cell-based assays suitable for detection of known molecular biomarkers in cancer using spherical cancer models (multicellular tumor spheroids and tissue-derived tumor spheres). We validate our procedures by detecting colorectal cancer (CRC) biomarkers both in patient-derived cultures and cell-lines (HCT116, HT29, LS174T). We expanded the study's scope to the analysis of specific CRC transcriptional and proteomic signatures in 3D. To this end, we employ primary cultures derived from CRC patients, and cell lines. Our methodology takes advantage of multiplex antibody microarrays to detect proteomic levels of 10 metalloproteinases of which MMP-13 and MMP- 9, as well as of surface proteins known to be involved in colorectal cancer invasion. Alterations in oxygen tension are milestones in tumor progression, and low oxygen tension (hypoxia) has been demonstrated in CRC. The Hypoxia Inducible Factor (HIF) transcription factor family is central to the response to hypoxic stress and is correlated with CRC carcinogenesis, invasion, VEGF expression, and poorer diagnosis. To increase the relevance of our model we use low oxygen conditions (3-5% physioxia conditions), and atmospheric oxygen levels (normoxia conditions) finding relevant differences at the proteomic, transcriptional and miRNA levels (also verified by qRT-PCR). We test the spherical cancer models on microfluidics chip and assess their vessel invasion capacity in normoxia and hypoxia, finding a correlation between hypoxia stress response and augmented invasiveness. Additionally, we test cellular responses to standard drugs and vasculature invasive capacity by time-lapse microscopy on-chip.

We design, develop and validate cellular assays for human cancer 3D culture generation, long-term maintenance and analysis. We analyze and quantify cellular responses to cytotoxic drugs, image and quantify invasion in fluidic chambers, determine levels of apoptosis, cell proliferation.

We demonstrate that a combination of controllable assays can successfully be used for identification of biomarker expression and compound screening in preclinical studies. 3D models are, therefore, a cost-effective in vitro model capable of recapitulating reliably physiological responses.

Citation Format: Dora Sabino, Isabelle Fixe, Alexandra Foucher, Eric Mennesson, Nadia Normand. Time-lapse analysis of drug response and invasive capacity using patient-derived CRC microfluidic 3D cancer models under oxygen-controlled conditions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5034.