Abstract 450: Systems biology approach provides rationale for dual-targeted inhibition of BET and CHK1 in aggressive pediatric osteosarcoma
Authors: Bijangi-Vishehsaraei Khadijeh , Pandya Pankita , Lijun Cheng , Shan Tang , Sinn Anthony , Trowbridge Melissa , Coy Kathy , Hemenway Courtney , Bailey Barbara , Shannon Harlan , Ding Jixin , Dobrota Erika , Saadatzadeh M. Reza , Elmi Adily , Shultz Jeremiah , Murray Mary , Marshall Mark , Ferguson Michael , Bertrand Todd , Wurtz L. Daniel , Batra Sandeep , Li Lang , Renbarger Jamie , Pollok Karen 
Published: September 2020

Abstract
Patients with aggressive osteosarcoma (OS) have poor prognosis due in part to copy number variations (CNVs) that contribute to dysregulation of gene expression (GE) and therapeutic resistance. The objective of the present study was to utilize the TARGET database to integrate CNV and corresponding GE with poor prognosis in pediatric OS (n=85) followed by functional validation of prioritized targets. Cox regression analysis indicated that CNVs in 2642 genes correlated with relapse risk in pediatric OS. Furthermore, the top 10 genes with CNVs significantly associated with increased risk for relapse were present on chromosome 8. The MYC and RAD21 copy number gain (MYC-RAD21 CNV+) located on chromosome 8q correlated with increased GE and poor survival in &gt;90% of the relapsed patients. Based on network analysis, the MYC-RAD21 CNV+ was prioritized for development of targeted therapy. MYC, an oncogenic driver of OS growth, can be indirectly inhibited by bromodomain and extra-terminal domain inhibitors (BETi). RAD21 expression has been associated with increased sensitivity to cell cycle checkpoint kinase 1 inhibitors (CHK1i) in melanoma. Additionally, mechanistic links exist between MYC and CHK1, especially during replication stress. Our hypothesis was that the MYC-RAD21 CNV+ serves as a biomarker of poor prognosis and therapeutic response to BETi+CHK1i therapy. Cell growth response to BETi and CHK1i was evaluated in MYC-RAD21 CNV+ pediatric OS cell lines and a patient-derived xenograft (PDX)-derived xenoline (TT2-77). OS lines (G292, MG63, U2OS, and TT2-77) were highly sensitive to single agent BETi/OTX-015, CHK1i/ SRA737 or CHK1i/LY2606368 at clinically relevant concentrations. Combination index and Bliss independence analysis indicated that BETi+CHK1i did not result in synergistic or additive inhibition of growth at clinically relevant concentrations. However, in OS lines Saos2 and Saos2-LM7 BETi+CHK1i resulted in additive to synergistic inhibition of growth at multiple dose-ratios and at clinically relevant concentrations. In the TT2-77 PDX, whole genome sequencing indicated that the original OS biopsy and the TT2-77 PDX generated from a resection sample harbor the MYC-RAD21 CNV+ (4 copies/amplicon). PDX tumor fragments were implanted into the flank of immunodeficient NOD/SCID/IL2Rγ mice. Once tumor volumes reached 100-150 mm3, mice were randomized and treated with four 5-day cycles of BETi/OTX-015 and/or CHK1i/SRA737. BETi+CHK1i significantly decreased TT2-77 growth, increased probability of survival, and was well tolerated. BETi+CHK1i is a promising therapeutic approach for treatment of relapsed pediatric MYC-RAD21 CNV+ OS. It is possible that MYC, BETs, RAD21 and CHK1 protein levels could dictate sensitivity to combination BETi+CHK1i independent of MYC-RAD2 CNV+ status. Studies are in progress to identify responder versus non-responder signatures in OS.
Citation Format: Khadijeh Bijangi-Vishehsaraei, Pankita Pandya, Cheng Lijun, Tang Shan, Anthony Sinn, Melissa Trowbridge, Kathy Coy, Courtney Hemenway, Barbara Bailey, Harlan Shannon, Jixin Ding, Erika Dobrota, M. Reza Saadatzadeh, Adily Elmi, Jeremiah Shultz, Mary Murray, Mark Marshall, Michael Ferguson, Todd Bertrand, L. Daniel Wurtz, Sandeep Batra, Lang Li, Jamie Renbarger, Karen Pollok. Systems biology approach provides rationale for dual-targeted inhibition of BET and CHK1 in aggressive pediatric osteosarcoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 450.