Gene expression and mutation-guided synthetic lethality eradicates proliferating and quiescent leukemia cells
Margaret Nieborowska-Skorska, … , Stephen M. Sykes, Tomasz Skorski
Margaret Nieborowska-Skorska, … , Stephen M. Sykes, Tomasz Skorski
Published May 8, 2017
Citation Information: J Clin Invest. 2017;127(6):2392-2406. https://doi.org/10.1172/JCI90825.
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Research Article Hematology

Gene expression and mutation-guided synthetic lethality eradicates proliferating and quiescent leukemia cells

Abstract

Quiescent and proliferating leukemia cells accumulate highly lethal DNA double-strand breaks that are repaired by 2 major mechanisms: BRCA-dependent homologous recombination and DNA-dependent protein kinase–mediated (DNA-PK–mediated) nonhomologous end-joining, whereas DNA repair pathways mediated by poly(ADP)ribose polymerase 1 (PARP1) serve as backups. Here we have designed a personalized medicine approach called gene expression and mutation analysis (GEMA) to identify BRCA- and DNA-PK–deficient leukemias either directly, using reverse transcription-quantitative PCR, microarrays, and flow cytometry, or indirectly, by the presence of oncogenes such as BCR-ABL1. DNA-PK–deficient quiescent leukemia cells and BRCA/DNA-PK–deficient proliferating leukemia cells were sensitive to PARP1 inhibitors that were administered alone or in combination with current antileukemic drugs. In conclusion, GEMA-guided targeting of PARP1 resulted in dual cellular synthetic lethality in quiescent and proliferating immature leukemia cells, and is thus a potential approach to eradicate leukemia stem and progenitor cells that are responsible for initiation and manifestation of the disease. Further, an analysis of The Cancer Genome Atlas database indicated that this personalized medicine approach could also be applied to treat numerous solid tumors from individual patients.

Authors

Margaret Nieborowska-Skorska, Katherine Sullivan, Yashodhara Dasgupta, Paulina Podszywalow-Bartnicka, Grazyna Hoser, Silvia Maifrede, Esteban Martinez, Daniela Di Marcantonio, Elisabeth Bolton-Gillespie, Kimberly Cramer-Morales, Jaewong Lee, Min Li, Artur Slupianek, Daniel Gritsyuk, Sabine Cerny-Reiterer, Ilona Seferynska, Tomasz Stoklosa, Lars Bullinger, Huaqing Zhao, Vera Gorbunova, Katarzyna Piwocka, Peter Valent, Curt I. Civin, Markus Muschen, John E. Dick, Jean C.Y. Wang, Smita Bhatia, Ravi Bhatia, Kolja Eppert, Mark D. Minden, Stephen M. Sykes, Tomasz Skorski

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Figure 7

PARP1i exerted a therapeutic effect in mice bearing BRCA/DNA-PK–deficient PLXs.

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PARP1i exerted a therapeutic effect in mice bearing BRCA/DNA-PK–deficien...
(A) Experimental design: NSG mice were injected with PLX cells and treated with diluents (control), doxorubicin + ara-C (DA), BMN673, or DA + BMN673 (8 mice/group). Human CD45+ (hCD45+) cells were detected in peripheral blood leukocytes (PBLs) 1 week after the treatment, and survival was determined. (B) Microarray profile of the indicated genes in AML (top plot) and B-ALL (middle and bottom plots) PLXs. Each circle represents an individual PLX; error bars show SD of the mean. Data from PLXs used for further experiments are marked in blue, gray, and red, indicating lower than average, average, and higher than average expression levels, respectively, of the indicated genes in BRCA/DNA-PK–deficient (BRCA/DNA-PK-D) AML and B-ALL (top and middle plots, respectively) and in BRCA/DNA-PK–proficient (BRCA/DNA-PK-p) B-ALL (bottom plot) cells. (C) Living cells from individual PLXs treated in vitro (triplicate experiment) with diluents (green), DA (purple), BMN673 (blue), or DA + BMN673 (black). (D) Representative plots of PBLs from treated mice (n = 6/group); percentage of hCD45+ cells (red dots) is indicated. *P ≤ 0.001, **P ≤ 0.001 in comparison with control and DA or BMN673, respectively, using Student’s t test adjusted for multiple comparisons. ***P < 0.03 in comparison with DA- or BMN673-treated mice using 2-way ANOVA. (E) Survival curves and MST of PLX mice treated with diluents (green), DA (purple), BMN673 (blue), or DA + BMN673 (black). * P < 0.003, **P < 0.003 in comparison with control and DA or BMN673, respectively, using Kaplan-Meier log-rank test.