Topoisomerase levels determine chemotherapy response in vitro and in vivo
Abstract
Topoisomerase poisons are chemotherapeutic agents that are used extensively for treating human malignancies. These drugs can be highly effective, yet tumors are frequently refractory to treatment or become resistant upon tumor relapse. Using a pool-based RNAi screening approach and a well characterized mouse model of lymphoma, we explored the genetic basis for heterogeneous responses to topoisomerase poisons in vitro and in vivo. These experiments identified Top2A expression levels as major determinants of response to the topoisomerase 2 poison doxorubicin and showed that suppression of Top2A produces resistance to doxorubicin in vitro and in vivo. Analogously, using a targeted RNAi approach, we demonstrated that suppression of Top1 produces resistance to the topoisomerase 1 poison camptothecin yet hypersensitizes cancer cells to doxorubicin. Importantly, lymphomas relapsing after treatment display spontaneous changes in topoisomerase levels as predicted by in vitro gene knockdown studies. These results highlight the utility of pooled shRNA screens for identifying genetic determinants of chemotherapy response and suggest strategies for improving the effectiveness of topoisomerase poisons in the clinic.
Acknowledgments.
We thank Lidia Nascimento, Beth Miller, and Stephanie Muller (CSHL) for analysis of shRNA representation by DNA sequencing, Carmelita Bautista (CSHL) for Chk2 antibodies, Holly Thompson for H2AX analysis, and Luke Gilbert for help with sensitization studies. We thank Mona Spector for critical reading and editing of the manuscript and members of the Lowe and Hannon laboratories for helpful advice and discussions. D.J.B. is an Engelhorn Scholar of the Watson School of Biological Sciences (CSHL). W.X. is in the Molecular and Cellular Biology graduate program at Stony Brook University (Stony Brook, NY). This work was supported by Alan and Edith Seligson (to L.Z.), the V Foundation for Cancer Research (to M.T.H.), a Mouse Models of Human Cancer Consortium grant, a program project grant from the National Cancer Institute (to G.J.H. and S.W.L.), and the Don Monti Memorial Research Foundation.
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Cover image: The Cubozoan jellyfish Tripedalia cystophora has complex, camera-like eyes. Zbynek Kozmik et al., in the article on pages 8989–8993, show that T. cystophora eyes form from genetic building blocks typical of vertebrates. Vision appears to have evolved similarly in these distantly separated animal phyla. Image courtesy of Zbynek Kozmik.
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© 2008 by The National Academy of Sciences of the USA. Freely available online through the PNAS open access option.
Submission history
Received: October 21, 2007
Published online: July 1, 2008
Published in issue: July 1, 2008
Keywords
Acknowledgments
We thank Lidia Nascimento, Beth Miller, and Stephanie Muller (CSHL) for analysis of shRNA representation by DNA sequencing, Carmelita Bautista (CSHL) for Chk2 antibodies, Holly Thompson for H2AX analysis, and Luke Gilbert for help with sensitization studies. We thank Mona Spector for critical reading and editing of the manuscript and members of the Lowe and Hannon laboratories for helpful advice and discussions. D.J.B. is an Engelhorn Scholar of the Watson School of Biological Sciences (CSHL). W.X. is in the Molecular and Cellular Biology graduate program at Stony Brook University (Stony Brook, NY). This work was supported by Alan and Edith Seligson (to L.Z.), the V Foundation for Cancer Research (to M.T.H.), a Mouse Models of Human Cancer Consortium grant, a program project grant from the National Cancer Institute (to G.J.H. and S.W.L.), and the Don Monti Memorial Research Foundation.
Notes
This article contains supporting information online at www.pnas.org/cgi/content/full/0803513105/DCSupplemental.
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The authors declare no conflict of interest.
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