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PDF file 129K, Supplementary Methods provide additional information on experimental techniques and data analysis. Supplementary Legends describe supplementary figures S1-S10 and supplementary table S1

PDF file 3500K, Potency of KU60648 across 94 cancer cell lines (Figure S1), comparison of KU60648-sensitivity in mutant vs. wildtype lines for 9 exemplary DNA repair-associated genes (Figure S2), MSH3 variants and Sanger sequencing of BRCA1 (Figure S3), cell cycle analysis of 9 cell lines under KU60648 exposure (Figure S4), analysis of synergistic effects between KU60648 and doxorubicin in six cell lines (Figure S5), validation of PRKDC knockdown efficiency (Figure S6), analysis of XRCC5 knockdown cells (Figure S7), analysis of MSH3 knockout cells (Figure S8), monitoring of DNA double-strand break repair in 3 cell lines (Figure S9), waterfall plots of tumor volumes after 11-day treatment with KU60648 (Figure S10)

PDF file 37K, List of all cell lines included in this study. Cell line name, GI50 of KU60648 from initial cell line screen, sensitivity classification as well as mutation annotation source are recorded

In this file we show additional data which support the main findings in our manuscript, e.g. an overview of all immunofluorescent and histological images, population doubling of the cell lines used, additional in vivo data on etoposide and cisplatin treatment and in vivo data of the p53-proficiet setting.

Defects in maintaining genome integrity are a hallmark of cancer. The DNA damage response kinase ATM is frequently mutated in human cancer, but the significance of these events to chemotherapeutic efficacy has not been examined deeply in whole organism models. Here we demonstrate that bi-allelic Atm deletion in mouse models of Kras-mutant lung adenocarcinoma does not affect cisplatin responses. In marked contrast, Atm-deficient tumors displayed an enhanced response to the topoisomerase-II poison etoposide. Moreover, Atm-deficient cells and tumors were sensitive to the PARP inhibitor olaparib. This actionable molecular addiction to functional PARP1 signaling was preserved in models that were proficient or deficient in p53, resembling standard or high-risk genetic constellations, respectively. Atm deficiency also markedly enhanced sensitivity to the ATR inhibitor VE-822. Taken together, our results provide a functional rationale to profile human tumors for disabling ATM mutations, particularly given their impact on PARP1 and ATR inhibitors. Cancer Res; 77(11); 3040–56. ©2017 AACR.

This file contains a list of additional methods used in this study, e.g. in situ hybridisation, PCR, immunoblotting, apoptotic assay, population doubling, and statistical analysis.

Summary: Cdkn1a, which encodes p21, functions as a major route for p53-mediated cell-cycle arrest. However, the consequence of Cdkn1a gene dosage on tumor suppression has not been systematically investigated. Here, we employed BAC transgenesis to generate a Cdkn1aSUPER mouse, which harbors an additional Cdkn1a allele within its natural genomic context. We show that these mice display enhanced cell-cycle arrest and reduced apoptosis in response to genotoxic stress. Furthermore, using a chemically induced skin cancer model and an autochthonous Kras-driven lung adenocarcinoma model, we show that Cdkn1aSUPER mice display a cancer protection phenotype that is indistinguishable from that observed in Tp53SUPER animals. Moreover, we demonstrate that Tp53 and Cdkn1a cooperate in mediating cancer resistance, using a chemically induced fibrosarcoma model. Overall, our Cdkn1aSUPER allele enabled us to assess the contribution of Cdkn1a to Tp53-mediated tumor suppression. : Torgovnick et al. create a mouse model, carrying a third copy of Cdkn1a (p21), which shows enhanced cell-cycle arrest capacity and protection against DNA damage-induced apoptosis. The Cdkn1aSUPER animals display delayed epithelial regeneration and a robust cancer resistance phenotype, highlighting the importance of p21 in p53-dependent tumor suppression. Keywords: Cdkn1a, p21, p53, mouse model, cancer, tumor suppressor, cell cycle arrest, apoptosis, cancer protection

Cdkn1a, which encodes p21, functions as a major route for p53-mediated cell-cycle arrest. However, the consequence of Cdkn1a gene dosage on tumor suppression has not been systematically investigated. Here, we employed BAC transgenesis to generate a Cdkn1a