Proof of concept on the role of ex vivo lung cancer spheroids, cytokines expression and PBMCs profiling in monitoring disease history and response to treatments

Abstract

Background: In the era of precision medicine, cancer treatment strategies are aimed to be patient-tailored. Here, we report a translational study conducted on a NSCLC patient where the potentiality of ex vivo spheroidal cultures and peripheral blood biomarkers were investigated to analyze correspondence with treatment response. Methods: We developed a protocol to generate ex vivo spheroidal cultures from patient's surgical sample that has been used to assess drug sensitivity through cell proliferation MTT assay. Primary tumor tissue, patient-derived spheroids and patient's circulating tumor DNA (ctDNA) were characterized through immunohistochemistry (IHC), immunofluorescence and next generation sequencing (NSG) analysis. Tumor infiltrating lymphocytes (TILs) from surgical specimen were analyzed by FACS and the analysis of TCR repertoire was conducted using Spectratyping technique. Moreover, peripheral blood samples collected at different time points underwent qPCR analysis to assess cytokines expression and flow cytometry to study peripheral blood mononuclear cells (PBMCs). All these data were correlated with clinical and radiological evaluations. Results: Immunohistochemistry, immunofluorescence, NGS analysis and TCR repertoire assay showed elevated concordance among primary tumor tissue, spheroids and ctDNA. Cisplatin-based chemotherapy and anti-PD-1 treatment sensitivity assessed in spheroidal cultures allowed us to anticipate patient response to chemo- and immunotherapy. Furthermore, circulating cytokines expression levels and lymphocytes subpopulations profiling correlated with clinical and radiological response to first line anti-PD-1 therapy. Conclusions: This approach demonstrated the feasibility of using spheroidal cultures, cytokines expression levels and PBMCs profiling to follow patient's disease history and response to treatment in parallel with clinical and radiological evaluation. Legal entity responsible for the study: Università degli Studi della Campania Luigi Vanvitelli. Funding: Has not received any funding. Disclosure: F. Ciardiello: Advisory / Consultancy, Research grant / Funding (institution): Roche; Advisory / Consultancy, Research grant / Funding (institution): Bayer; Advisory / Consultancy, Research grant / Funding (institution): Amgen; Advisory / Consultancy: Merck; Advisory / Consultancy: Pfizer; Advisory / Consultancy: Sanofi; Advisory / Consultancy: Servier; Advisory / Consultancy: BMS; Advisory / Consultancy: Cellgene; Advisory / Consultancy: Lilly; Research grant / Funding (institution): AstraZeneca; Research grant / Funding (institution): Ipsen. F. Morgillo: Advisory / Consultancy: MSD; Advisory / Consultancy: Lilly; Research grant / Funding (institution): AstraZeneca. All other authors have declared no conflicts of interest.