Background: Drugs targeting the spindle assembly checkpoint (SAC), such as inhibitors of Aurora kinase B (AURKB) and dual specific protein kinase TTK, are in different stages of clinical development. However, cell response to SAC abrogation is poorly understood and there are no markers for patient selection. Methods: A panel of 53 tumor cell lines of different origins was used. The effects of drugs were analyzed by MTT and flow cytometry. Copy number status was determined by FISH and Q-PCR; mRNA expression by nCounter and RT-Q-PCR and protein expression by Western blotting. CRISPR-Cas9 technology was used for gene knock-out (KO) and a doxycycline-inducible pTRIPZ vector for ectopic expression. Finally, in vivo experiments were performed by implanting cultured cells or fragments of tumors into immunodeficient mice. Results: Tumor cells and patient-derived xenografts (PDXs) sensitive to AURKB and TTK inhibitors consistently showed high expression levels of BH3-interacting domain death agonist (BID), while cell lines and PDXs with low BID were uniformly resistant. Gene silencing rendered BID-overexpressing cells insensitive to SAC abrogation while ectopic BID expression in BID-low cells significantly increased sensitivity. SAC abrogation induced activation of CASP-2, leading to cleavage of CASP-3 and extensive cell death only in presence of high levels of BID. Finally, a prevalence study revealed high BID mRNA in 6% of human solid tumors. Conclusions: The fate of tumor cells after SAC abrogation is driven by an AURKB/ CASP-2 signaling mechanism, regulated by BID levels. Our results pave the way to clinically explore SAC-targeting drugs in tumors with high BID expression.
Bibliografisk noteFunding Information:
I.S. has received fees for consultancy or advisory roles from Roche, Novartis, Boehringer Ingelheim and Takeda. N.R. has received fees for consultancy or advisory roles from Merck Sharp & Dohme Corp, Bristol-Myers Squibb and Pfizer. A.A.H. has received fees for consultancy or advisory roles from Bristol-Myers Squibb, Roche, Merck Sharp & Dohme Corp and Lilly. J.T., S.T., R.W., N.F., G.G., M.X., P.S., J.S., G.F., A.G., U.P. and J.U. are current or former AstraZeneca employees and shareholders. M.A.M.V. has received research funding from Astra Zeneca, In3Bio and Merck Healthcare KGaA. The rest of the authors declare no competing interests.