Supplementary Data from GAS6/AXL Inhibition Enhances Ovarian Cancer Sensitivity to Chemotherapy and PARP Inhibition through Increased DNA Damage and Enhanced Replication Stress

Mullen, Mary M.; Lomonosova, Elena; Toboni, Michael D.; Oplt, Alyssa; Cybulla, Emily; Blachut, Barbara; Zhao, Peinan; Noia, Hollie; Wilke, Daniel; Rankin, Erinn B.; Kuroki, Lindsay M.; Hagemann, Andrea R.; Hagemann, Ian S.; McCourt, Carolyn K.; Thaker, Premal H.; Mutch, David G.; Powell, Matthew A.; Mosammaparast, Nima; Vindigni, Alessandro; Fuh, Katherine C.

doi:10.1158/1541-7786.22527323.v1

Supplementary Data from GAS6/AXL Inhibition Enhances Ovarian Cancer Sensitivity to Chemotherapy and PARP Inhibition through Increased DNA Damage and Enhanced Replication Stress

https://doi.org/10.1158/1541-7786.22527323

journal contribution

posted on 2023-04-03, 20:00 authored by Mary M. Mullen, Elena Lomonosova, Michael D. Toboni, Alyssa Oplt, Emily Cybulla, Barbara Blachut, Peinan Zhao, Hollie Noia, Daniel Wilke, Erinn B. Rankin, Lindsay M. Kuroki, Andrea R. Hagemann, Ian S. Hagemann, Carolyn K. McCourt, Premal H. Thaker, David G. Mutch, Matthew A. Powell, Nima Mosammaparast, Alessandro Vindigni, Katherine C. Fuh

<p>Supplementary Table S1. Comprehensive radiologic, surgical, and pathologic scoring system used to evaluate tumor response to neoadjuvant chemotherapy. Supplementary Table S2. Summary of antibodies used. Supplementary Table S3. Disease characteristics of ovarian cancer patients receiving neoadjuvant chemotherapy and undergoing interval cytoreductive surgery stratified by chemoresponse. Supplementary Table S4. Disease characteristics of ovarian cancer patients receiving neoadjuvant chemotherapy and undergoing interval cytoreductive surgery stratified by high and low serum GAS6 expression prior to neoadjuvant chemotherapy. Supplementary Figure S1. RAD51 foci formation in irradiated ovarian cancer cells. Supplementary Figure S2. GAS6 and AXL expression in four ovarian cancer cell lines. Supplementary Figure S3. Mean combination indexes and dose response curves for paclitaxel and AVB-500 in three ovarian cancer cell lines. Supplementary Figure S4. GAS6/AXL inhibitor AVB-500 improves response to carboplatin and decreases tumor burden in PDX mouse model of ovarian cancer. Supplementary Figure S5. GAS6/AXL inhibitor AVB-500 decreases GAS6 and pAKT expression in mouse tumors. Supplementary Figure S6. Cleaved Caspase 3 levels in OVCAR8 ovarian cancer cells at 24 and 48 hours of treatment. Supplementary Figure S7. Treatment with AVB-500 increases DNA damage. Supplementary Figure S8. Treatment with AVB-500 decreases activation of AKT and does not affect protein expression of 53BP1, AXL, or RAD51. Supplementary Figure S9. Treatment with carboplatin plus AVB-500 decreases pATM and increases pCHK1. Supplementary Figure S10. Treatment with AVB-500 decreases RPA binding and does not affect the cell cycle. Supplementary Figure S11. Treatment with AVB-500 in addition to olaparib increases DNA damage and decreases homologous recombination. Supplementary Figure S12. Proposed mechanism of AVB-500 and carboplatin increasing DNA damage through replication fork perturbation.</p>

Funding

Reproductive Scientist Development Program award

NCI F30 fellowship

Burroughs Wellcome Fund Physician-Scientist Institutional Award

History

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DOI - Is supplement to GAS6/AXL Inhibition Enhances Ovarian Cancer Sensitivity to Chemotherapy and PARP Inhibition through Increased DNA Damage and Enhanced Replication Stress

ARTICLE ABSTRACT

Over 80% of women with high-grade serous ovarian cancer (HGSOC) develop tumor resistance to chemotherapy and die of their disease. There are currently no FDA-approved agents to improve sensitivity to first-line platinum- and taxane-based chemotherapy or to PARP inhibitors. Here, we tested the hypothesis that expression of growth arrest–specific 6 (GAS6), the ligand of receptor tyrosine kinase AXL, is associated with chemotherapy response and that sequestration of GAS6 with AVB-S6–500 (AVB-500) could improve tumor response to chemotherapy and PARP inhibitors. We found that GAS6 levels in patient tumor and serum samples collected before chemotherapy correlated with ovarian cancer chemoresponse and patient survival. Compared with chemotherapy alone, AVB-500 plus carboplatin and/or paclitaxel led to decreased ovarian cancer-cell survival in vitro and tumor burden in vivo. Cells treated with AVB-500 plus carboplatin had more DNA damage, slower DNA replication fork progression, and fewer RAD51 foci than cells treated with carboplatin alone, indicating AVB-500 impaired homologous recombination (HR). Finally, treatment with the PARP inhibitor olaparib plus AVB-500 led to decreased ovarian cancer-cell survival in vitro and less tumor burden in vivo. Importantly, this effect was seen in HR-proficient and HR-deficient ovarian cancer cells. Collectively, our findings suggest that GAS6 levels could be used to predict response to carboplatin and AVB-500 could be used to treat platinum-resistant, HR-proficient HGSOC. GAS6/AXL is a novel target to sensitize ovarian cancers to carboplatin and olaparib. Additionally, GAS6 levels can be associated with response to carboplatin treatment.