ATR inhibitors synergize with mitomycin C to enhance cytotoxicity in patient-derived non-muscle invasive bladder cancer organoids

Abstract

Background and ObjectiveThere is a high recurrence rate in non-muscle invasive bladder cancer (NMIBC) patients treated with intravesical chemotherapy or BCG, indicating the need for novel treatment options. Synergy between DNA damage response inhibitors and chemotherapy has been shown for different solid tumors. In this study, we explore whether combining chemotherapeutic agents with Ataxia telangiectasia and Rad3-related (ATR) kinase inhibitors is a suitable strategy to reduce recurrence in NMIBC. MethodsNMIBC patient-derived organoids (PDOs; n=6) were exposed to mitomycin C (MMC), gemcitabine, or epirubicin for 2h, to mimic intravesical instillation, and subsequently to ATR inhibitors (berzosertib, ceralasertib, tuvusertib) for 72h. Cell viability and PDO regrowth potential was determined by microscopy and CellTiter-Glo(R) luminescence assays, during a period of 6 weeks post-treatment. Key Findings and LimitationsPDO viability was severely impaired (1-22% viable cells at t=6 weeks) after combination treatment with MMC and ATR inhibitors. This effect was observed directly after treatment and maintained during 6 weeks after drug exposure. In contrast, PDOs treated with only chemotherapy or ATR inhibitors showed similar proliferation rates as untreated controls in week 6. LimitationsPDO models were all chemotherapy naive (either BCG or no pre-treatment). It would be interesting to confirm these findings in MMC-resistant models. Conclusions and Clinical ImplicationsThis study demonstrates synergy between different ATR inhibitors and MMC in a panel of six NMIBC PDOs. We propose that combining intravesical chemotherapy instillations with DNA damage response inhibitors should be further evaluated as a promising strategy to reduce recurrence rates for NMIBC. Plain English summaryIn this report we looked at new treatment options for non-muscle invasive bladder cancer. We discovered that tumor cells are killed very efficiently if chemotherapy is combined with drugs that prevent cells from repairing DNA damage.