Patient-derived organoids demonstrate synergistic effect of co-targeting Aurora kinase and prosurvival BCL2 family proteins

Many melanomas are resistant to standard-of-care immune checkpoint blockade and inhibitors of mutated BRAF. New effective therapies that work via immune and BRAF-independent mechanisms are urgently needed. We have previously demonstrated that an inhibitor of mitotic kinase Aurora A (AURKAi) can induce a distinct type of stable cell cycle known as senescence in melanoma tumors independent of their genetic background. Here we hypothesized that combination of AURKAi with a drug that selectively kills senescent cells, such as BCL2 inhibitor (BCLi) navitoclax, will be synthetically lethal to melanoma cells. Melanoma patient-derived organoids (PDOs) were used to test efficacy of AURKAi and BCLi combination treatment. To generate organoids, patient-derived tumor cells were cultured ex vivo in 3D matrix. Drug response was evaluated using fluorescent viability assay. Fifteen out of twenty PDOs were highly sensitive to AURKAi and BCLi combination treatment, exhibiting robust induction of cell death. No cell death was observed when either drug was added individually. NextGen DNA sequencing indicated that all sensitive PDOs retained wild-type TP53, while resistant tumors had mutations in this gene. Knockout or knockdown of TP53 abrogated AURKAi and BCLi-induced cell death in melanoma cells, suggesting that transcription factor p53 encoded by TP53 is a key mediator of drug response. Mechanistically, p53 was induced by AURKAi treatment independent of BCLi. However, single-agent AURKAi treatment resulted in activation of p53-mediated cell cycle arrest based on increased expression of CDK inhibitor p21 and decreased expression of proliferation markers. In contrast, addition of BCLi to AURKAi activated proapoptotic p53 target BAX and induced cleavage of PARP and caspases 3, 7, and 8. Treatment with pan-caspase inhibitor abrogated cell death, suggesting caspase-dependent apoptosis. Electron microscopy revealed morphologic signs of apoptosis in combination-treated cells, including cell blebbing, apoptotic bodies, and nuclear fragmentation. This suggests that BCLi shifts cell fate decision in AURKAi-treated cells towards apoptosis. In conclusion, combination treatment with AURKAi and BCl2i induced p53-dependent apoptosis. Preclinical evaluation of this drug combination in PDO model demonstrated robust efficacy against melanomas with wild-type p53. Since p53 mutations are relatively rare in melanoma, a combination of AURKAi and BCLi presents a promising therapeutic option for this deadly disease. Citation Format: Vijaya Bharti, Ashlyn Blevins, Vivian L. Weiss, Sheau C Chen, Roman Uzhachenko, Ann Richmond, Anna E. Vilgelm. Patient-derived organoids demonstrate synergistic effect of co-targeting Aurora kinase and prosurvival BCL2 family proteins [abstract]. In: Proceedings of the AACR Special Conference on the Evolving Landscape of Cancer Modeling; 2020 Mar 2-5; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2020;80(11 Suppl):Abstract nr B20.