Multi-Cell Type Glioblastoma Tumor Spheroids for Evaluating Sub-Population-Specific Drug Response

Hemamylammal Sivakumar; Mahesh Devarasetty; David E Kram; Roy E Strowd; Aleksander Skardal

doi:10.3389/fbioe.2020.538663

Multi-Cell Type Glioblastoma Tumor Spheroids for Evaluating Sub-Population-Specific Drug Response

Front Bioeng Biotechnol. 2020 Sep 15:8:538663. doi: 10.3389/fbioe.2020.538663. eCollection 2020.

Authors

Hemamylammal Sivakumar^{1

2

3}, Mahesh Devarasetty³, David E Kram^{4

5}, Roy E Strowd^{5

6}, Aleksander Skardal^{1

2

3

5

7

8

9}

Affiliations

¹ Department of Biomedical Engineering, The Ohio State University, Columbus, OH, United States.
² The Ohio State University and Arthur G. James Comprehensive Cancer Center, Columbus, OH, United States.
³ Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, NC, United States.
⁴ Section of Pediatric Hematology and Oncology, Department of Pediatrics, Wake Forest Baptist Medical Center, Medical Center Boulevard, Winston-Salem, NC, United States.
⁵ Comprehensive Cancer Center at Wake Forest Baptist Medical, Winston-Salem, NC, United States.
⁶ Department of Neurology, Wake Forest Baptist Medical Center, Winston-Salem, NC, United States.
⁷ Virginia Tech-Wake Forest School of Biomedical Engineering and Sciences, Wake Forest School of Medicine, Winston-Salem, NC, United States.
⁸ Department of Cancer Biology, Wake Forest School of Medicine, Winston-Salem, NC, United States.
⁹ Department of Molecular Medicine and Translational Science, Wake Forest School of Medicine, Winston-Salem, NC, United States.

Abstract

Glioblastoma (GBM) is a lethal, incurable form of cancer in the brain. Even with maximally aggressive surgery and chemoradiotherapy, median patient survival is 14.5 months. These tumors infiltrate normal brain tissue, are surgically incurable, and universally recur. GBMs are characterized by genetic, epigenetic, and microenvironmental heterogeneity, and they evolve spontaneously over time and as a result of treatment. However, tracking such heterogeneity in real time in response to drug treatments has been impossible. Here we describe the development of an in vitro GBM tumor organoid model that is comprised of five distinct cellular subpopulations (4 GBM cell lines that represent GBM subpopulations and 1 astrocyte line), each fluorescently labeled with a different color. These multi-cell type GBM organoids are then embedded in a brain-like hyaluronic acid hydrogel for subsequent studies involving drug treatments and tracking of changes in relative numbers of each fluorescently unique subpopulation. This approach allows for the visual assessment of drug influence on individual subpopulations within GBM, and in future work can be expanded to supporting studies using patient tumor biospecimen-derived cells for personalized diagnostics.

Keywords: drug response; glioblastoma; organoid; spheroid; tumor heterogeneity.