Abstract
Malignant mesothelioma (MM) is an aggressive, fatal tumor strongly associated with asbestos exposure. There is an urgent need to improve MM patient outcomes and this requires functionally validated pre-clinical models. Mesothelioma-derived cell lines provide an essential and relatively robust tool and remain among the most widely used systems for candidate drug evaluation. Although a number of cell lines are commercially available, a detailed comparison of these commercial lines with freshly derived primary tumor cells to validate their suitability as pre-clinical models is lacking. To address this, patient-derived primary mesothelioma cell lines were established and characterized using complementary multidisciplinary approaches and bioinformatic analysis. Clinical markers of mesothelioma, transcriptional and metabolic profiles, as well as the status of p53 and the tumor suppressor genes CDKN2A and NF2, were examined in primary cell lines and in two widely used commercial lines. Expression of MM-associated markers, as well as the status of CDKN2A, NF2, the ‘gatekeeper' in MM development, and their products demonstrated that primary cell lines are more representative of the tumor close to its native state and show a degree of molecular diversity, thus capturing the disease heterogeneity in a patient cohort. Molecular profiling revealed a significantly different transcriptome and marked metabolic shift towards a greater glycolytic phenotype in commercial compared with primary cell lines. Our results highlight that multiple, appropriately characterised, patient-derived tumor cell lines are required to enable concurrent evaluation of molecular profiles versus drug response. Furthermore, application of this approach to other difficult-to-treat tumors would generate improved cellular models for pre-clinical evaluation of novel targeted therapies.
Highlights
Received 28.5.15; revised 04.11.15; accepted 16.11.15; Edited by G Melino; published online 19.2.16 important as MM is associated with chromosomal loss, deletions in CDKN2A, CDKN2B, and NF213–15 genes and mutations in BAP1 and CUL1.16 genomic instability limits the long-term usefulness of commercially available MM cell lines
Our data demonstrate that there is a significant difference between commercial and primary cell lines at the molecular level, including the transcriptome, expression of mesothelial markers and proteins associated with MM pathogenesis including p53, metabolic profile, status of the tumor suppressor genes CDKN2A and NF2 and their products
We examined the status of p16INK4A/p14ARF genetic loci in both primary and commercial mesothelioma cell lines (Figure 4a and b)
Summary
Received 28.5.15; revised 04.11.15; accepted 16.11.15; Edited by G Melino; published online 19.2.16 important as MM is associated with chromosomal loss, deletions in CDKN2A, CDKN2B, and NF213–15 genes and mutations in BAP1 and CUL1.16 genomic instability limits the long-term usefulness of commercially available MM cell lines. To develop more relevant pre-clinical models of MM, we established and characterised eight primary mesothelioma cell lines and employed a number of complementary multidisciplinary approaches to examine differences between these cell lines and two widely used commercial cell lines. Our data demonstrate that there is a significant difference between commercial and primary cell lines at the molecular level, including the transcriptome, expression of mesothelial markers and proteins associated with MM pathogenesis including p53, metabolic profile, status of the tumor suppressor genes CDKN2A and NF2 and their products. The commercial cell lines lack many key molecular features known to be associated with MM, whereas the eight primary cell lines more accurately recapitulate human disease, providing a superior model for pre-clinical evaluation of novel targeted therapies
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