Comparative Molecular Analysis of Cancer Behavior Cultured In Vitro, In Vivo, and Ex Vivo

Hum, Nicholas R.; Sebastian, Aimy; Gilmore, Sean F.; He, Wei; Martin, Kelly A.; Hinckley, Aubree; Dubbin, Karen R.; Moya, Monica L.; Wheeler, Elizabeth K.; Coleman, Matthew A.; Loots, Gabriela G.

doi:10.3390/cancers12030690

Comparative Molecular Analysis of Cancer Behavior Cultured In Vitro, In Vivo, and Ex Vivo

Journal Article · Sat Mar 14 00:00:00 EDT 2020 · Cancers (Basel)

DOI:https://doi.org/10.3390/cancers12030690· OSTI ID:1605484

; ; Gilmore, Sean F.; ; ; Hinckley, Aubree; ; ; Wheeler, Elizabeth K.; Coleman, Matthew A.;

Current pre-clinical models of cancer fail to recapitulate the cancer cell behavior in primary tumors primarily because of the lack of a deeper understanding of the effects that the microenvironment has on cancer cell phenotype. Transcriptomic profiling of 4T1 murine mammary carcinoma cells from 2D and 3D cultures, subcutaneous or orthotopic allografts (from immunocompetent or immunodeficient mice), as well as ex vivo tumoroids, revealed differences in molecular signatures including altered expression of genes involved in cell cycle progression, cell signaling and extracellular matrix remodeling. The 3D culture platforms had more in vivo-like transcriptional profiles than 2D cultures. In vivo tumors had more cells undergoing epithelial-to-mesenchymal transition (EMT) while in vitro cultures had cells residing primarily in an epithelial or mesenchymal state. Ex vivo tumoroids incorporated aspects of in vivo and in vitro culturing, retaining higher abundance of cells undergoing EMT while shifting cancer cell fate towards a more mesenchymal state. Cellular heterogeneity surveyed by scRNA-seq revealed that ex vivo tumoroids, while rapidly expanding cancer and fibroblast populations, lose a significant proportion of immune components. This study emphasizes the need to improve in vitro culture systems and preserve syngeneic-like tumor composition by maintaining similar EMT heterogeneity as well as inclusion of stromal subpopulations.

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Research Organization:: Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)

Sponsoring Organization:: USDOE; USDOE National Nuclear Security Administration (NNSA); USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division

Grant/Contract Number:: AC52-07NA27344

OSTI ID:: 1605484

Alternate ID(s):: OSTI ID: 1629048
OSTI ID: 1841860

Report Number(s):: LLNL-JRNL--782408; PII: cancers12030690

Journal Information:: Cancers (Basel), Journal Name: Cancers (Basel) Journal Issue: 3 Vol. 12; ISSN CANCCT; ISSN 2072-6694

Publisher:: MDPI AGCopyright Statement

Country of Publication:: Switzerland

Language:: English

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