Synergistic activity of ABT-263 and ONC201/TIC10 against solid tumor cell lines is associated with suppression of anti-apoptotic Mcl-1, BAG3, pAkt, and upregulation of pro-apoptotic Noxa and Bax cleavage during apoptosis

Francesca R Di Cristofano; Mara W Fong; Kelsey E Huntington; Benedito A Carneiro; Lanlan Zhou; Wafik S El-Deiry

Synergistic activity of ABT-263 and ONC201/TIC10 against solid tumor cell lines is associated with suppression of anti-apoptotic Mcl-1, BAG3, pAkt, and upregulation of pro-apoptotic Noxa and Bax cleavage during apoptosis

Am J Cancer Res. 2023 Jan 15;13(1):307-325. eCollection 2023.

Authors

Francesca R Di Cristofano^{1

2

3

4}, Mara W Fong^{1

2

3

4}, Kelsey E Huntington^{1

2

3

5

4}, Benedito A Carneiro^{1

2

6

4}, Lanlan Zhou^{1

2

3

4}, Wafik S El-Deiry^{1

2

3

5

7

6

4}

Affiliations

¹ Laboratory of Translational Oncology and Experimental Cancer Therapeutics, The Warren Alpert Medical School, Brown University Providence 02903, RI, USA.
² The Joint Program in Cancer Biology, Brown University and The Lifespan Health System Providence 02903, RI, USA.
³ Department of Pathology and Laboratory Medicine, The Warren Alpert Medical School, Brown University Providence 02903, RI, USA.
⁴ Legorreta Cancer Center at Brown University, The Warren Alpert Medical School, Brown University Providence 02903, RI, USA.
⁵ Pathobiology Graduate Program, The Warren Alpert Medical School, Brown University Providence 02903, RI, USA.
⁶ Hematology-Oncology Division, Department of Medicine, Rhode Island Hospital and Brown University Providence 02903, RI, USA.
⁷ Molecular and Cellular Biology Graduate Program, The Warren Alpert Medical School of Brown University Providence 02903, RI, USA.

PMID: 36777502
PMCID: PMC9906082

Abstract

A major underlying cause of the resistance of solid tumor cells to cancer therapy is the evasion of cell death following anti-cancer drug treatment. We explored the combination of TRAIL-inducing compound ONC201/TIC10 and Bcl-xL/Bcl-2 inhibitor ABT-263 to target the extrinsic and intrinsic apoptotic pathways, respectively, in solid tumor cell lines (N = 13) derived from different tissues (colon, prostate, lung, breast, ovary, bladder). We found an IC50 range of 0.83-20.10 μM for ONC201 and 0.06-14.75 μM for ABT-263 among the 13 cancer cell lines. We show that combination of ONC201 and ABT-263 produces a strong synergistic effect leading to tumor cell death, and that the combination is not toxic to human fibroblast cells. In OVCAR-3 ovarian cancer cells, 2.5 μM ONC201 and 1.25 μM ABT-263 yielded 37% and 27% inhibition of viability, respectively, while the combination of the two agents yielded 92% inhibition of viability, resulting in a high synergy score of 52; conversely, the same combination in the HFF-1 human fibroblast cells yielded 2.45% inhibition of viability and a synergy score of 6.92 (synergy scores were calculated using SynergyFinder; scores greater than 10 are considered synergistic). We also found that the combination of these two agents resulted in synergistic caspase activation and PARP cleavage consistent with induction of apoptosis. Combination therapy-induced cell death correlated with decreased levels of Mcl-1, BAG3, pAkt, and upregulation of Noxa along with Bax cleavage during apoptosis at 48 hours, and ATF4, TRAIL, and DR5 induction at 24 hours. There was some heterogeneity in the cell lines with regard to these responses. Our data provide evidence for synergy from the combination of ONC201 and ABT-263 against human solid tumor cell lines associated with alterations in cell death and pro-survival mediators. The combination of ONC201 and ABT-263 merits further exploration in vivo and in clinical trials against a variety of solid malignancies.

Keywords: ABT-263; Akt; Apoptosis; BAG3; Mcl-1; Noxa; ONC201; drug synergy; targeted therapy.