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

Abstract

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 ofmore » stromal subpopulations.« less

Authors:

Hum, Nicholas R. ;

• Search DOE PAGES for author "Hum, Nicholas R."
• Search DOE PAGES for ORCID "0000-0003-1605-3193"

Sebastian, Aimy ;

• Search DOE PAGES for author "Sebastian, Aimy"
• Search DOE PAGES for ORCID "0000-0002-7822-7040"

Gilmore, Sean F.; He, Wei ;

• Search DOE PAGES for author "He, Wei"
• Search DOE PAGES for ORCID "0000-0003-2795-0324"

Martin, Kelly A. ;

• Search DOE PAGES for author "Martin, Kelly A."
• Search DOE PAGES for ORCID "0000-0002-0025-994X"

Hinckley, Aubree; Dubbin, Karen R. ;

• Search DOE PAGES for author "Dubbin, Karen R."
• Search DOE PAGES for ORCID "0000-0002-9495-4766"

Moya, Monica L. ;

• Search DOE PAGES for author "Moya, Monica L."
• Search DOE PAGES for ORCID "0000-0002-6714-1700"

Wheeler, Elizabeth K.; Coleman, Matthew A.; Loots, Gabriela G.

• Search DOE PAGES for author "Loots, Gabriela G."
• Search DOE PAGES for ORCID "0000-0001-9546-5561"

Publication Date:

Sat Mar 14 00:00:00 EDT 2020

Research Org.:

Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)

Sponsoring Org.:

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

OSTI Identifier:

1605484

Alternate Identifier(s):

OSTI ID: 1629048; OSTI ID: 1841860

Report Number(s):

LLNL-JRNL-782408
Journal ID: ISSN 2072-6694; CANCCT; PII: cancers12030690

Grant/Contract Number:  

19-SI-003; AC52-07NA27344

Resource Type:

Published Article

Journal Name:

Cancers (Basel)

Additional Journal Information:

Journal Name: Cancers (Basel) Journal Volume: 12 Journal Issue: 3; Journal ID: ISSN 2072-6694

Publisher:

MDPI AG

Country of Publication:

Switzerland

Language:

English

Subject:

59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES