Replication stress induces mitotic death through parallel pathways regulated by WAPL and telomere deprotection

Mitotic catastrophe is a broad descriptor encompassing unclear mechanisms of cell death. Here we investigate replication stress-driven mitotic catastrophe in human cells and identify that replication stress principally induces mitotic death signalled through two independent pathways. In p53-compromised cells we find that lethal replication stress confers WAPL-dependent centromere cohesion defects that maintain spindle assembly checkpoint-dependent mitotic arrest in the same cell cycle. Mitotic arrest then drives cohesion fatigue and triggers mitotic death through a primary pathway of BAX/BAK-dependent apoptosis. Simultaneously, a secondary mitotic death pathway is engaged through non-canonical telomere deprotection, regulated by TRF2, Aurora B and ATM. Additionally, we find that suppressing mitotic death in replication stressed cells results in distinct cellular outcomes depending upon how cell death is averted. These data demonstrate how replication stress-induced mitotic catastrophe signals cell death with implications for cancer treatment and cancer genome evolution

Supplementary Data Figure S3 from Loss-of-Function but Not Gain-of-Function Properties of Mutant TP53 Are Critical for the Proliferation, Survival, and Metastasis of a Broad Range of Cancer Cells

Supplementary Figure S3 showing that removal of mutant TP53 does not impair the ability of diverse human cancer cell lines to adapt to conditions of stress</p

Supplementary Data Table S6 from Loss-of-Function but Not Gain-of-Function Properties of Mutant TP53 Are Critical for the Proliferation, Survival, and Metastasis of a Broad Range of Cancer Cells

Supplementary Table S6 showing the impact of removal of mutant BRAF in human cancer cell lines in vitro by CRISPR/Cas9 by mining the DepMap database</p

Supplementary Data Figure S27 from Loss-of-Function but Not Gain-of-Function Properties of Mutant TP53 Are Critical for the Proliferation, Survival, and Metastasis of a Broad Range of Cancer Cells

Supplementary Figure S27 showing that removal of mutant TP53 does not impact the in vitro response of human colon cancer derived organoids to cisplatin</p

Supplementary Data Figure S11 from Loss-of-Function but Not Gain-of-Function Properties of Mutant TP53 Are Critical for the Proliferation, Survival, and Metastasis of a Broad Range of Cancer Cells

Supplementary Figure S11 showing that removal of mutant TP53 does not cause a competitive growth disadvantage in diverse human cancer cell lines in culture</p

Supplementary Data Table S9 from Loss-of-Function but Not Gain-of-Function Properties of Mutant TP53 Are Critical for the Proliferation, Survival, and Metastasis of a Broad Range of Cancer Cells

Supplementary Table S9 showing the impact of removal of mutant KRAS in human cancer cell lines in vitro by RNAi by mining the DepMap database</p

Supplementary Data Figure S26 from Loss-of-Function but Not Gain-of-Function Properties of Mutant TP53 Are Critical for the Proliferation, Survival, and Metastasis of a Broad Range of Cancer Cells

Supplementary Figure S26 showing that removal of mutant TP53 does not impact in vivo tumor growth upon transplantation of WCB139T human colon cancer derived organoids into NSG mice</p

Supplementary Data Figure S28 from Loss-of-Function but Not Gain-of-Function Properties of Mutant TP53 Are Critical for the Proliferation, Survival, and Metastasis of a Broad Range of Cancer Cells

Supplementary Figure S28 showing that removal of mutant TP53 has no impact on the response of tumors derived from human colon cancer derived organoids to 5-FU in vivo</p

Supplementary Data Figure S9 from Loss-of-Function but Not Gain-of-Function Properties of Mutant TP53 Are Critical for the Proliferation, Survival, and Metastasis of a Broad Range of Cancer Cells

Supplementary Figure S9 showing that complete removal of mutant TP53 in Rael-BL cells does not impair survival and proliferation in culture</p

Supplementary Data Figure S11 from Loss-of-Function but Not Gain-of-Function Properties of Mutant TP53 Are Critical for the Proliferation, Survival, and Metastasis of a Broad Range of Cancer Cells

Supplementary Figure S11 showing that removal of mutant TP53 does not cause a competitive growth disadvantage in diverse human cancer cell lines in culture</p