Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Somatic deletions of genes regulating MSH2 protein stability cause DNA mismatch repair deficiency and drug resistance in human leukemia cells

Nature Medicine volume 17, pages 1298–1303 (2011)Cite this article

Subjects

Abstract

DNA mismatch repair enzymes (for example, MSH2) maintain genomic integrity, and their deficiency predisposes to several human cancers and to drug resistance. We found that leukemia cells from a substantial proportion of children (∼11%) with newly diagnosed acute lymphoblastic leukemia have low or undetectable MSH2 protein levels, despite abundant wild-type MSH2 mRNA. Leukemia cells with low levels of MSH2 contained partial or complete somatic deletions of one to four genes that regulate MSH2 degradation (FRAP1 (also known as MTOR), HERC1, PRKCZ and PIK3C2B); we also found these deletions in individuals with adult acute lymphoblastic leukemia (16%) and sporadic colorectal cancer (13.5%). Knockdown of these genes in human leukemia cells recapitulated the MSH2 protein deficiency by enhancing MSH2 degradation, leading to substantial reduction in DNA mismatch repair and increased resistance to thiopurines. These findings reveal a previously unrecognized mechanism whereby somatic deletions of genes regulating MSH2 degradation result in undetectable levels of MSH2 protein in leukemia cells, DNA mismatch repair deficiency and drug resistance.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Get just this article for as long as you need it

$39.95

Learn more

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Gene copy number loss and MSH2 protein expression in primary human leukemia cells.
Figure 2: Protein expression in primary leukemia cells with hemizygous deletions, treatment outcome and drug sensitivity according to leukemia cell MSH2 phenotype.
Figure 3: PRKCZ, PIK3C2B, HERC1 and FRAP1 inhibition and MSH2 stability.
Figure 4: Increase in PP2A activity through inhibition or knockdown of FRAP1, HERC1 or PIK3C2B with rescue by okadaic acid (OA) and the effects on MMR activity.

References

  1. Felton, K.E., Gilchrist, D.M. & Andrew, S.E. Constitutive deficiency in DNA mismatch repair. Clin. Genet. 71, 483–498 (2007).

    Article  CAS  Google Scholar 

  2. Fishel, R. et al. The human mutator gene homolog MSH2 and its association with hereditary nonpolyposis colon cancer. Cell 75, 1027–1038 (1993).

    Article  CAS  Google Scholar 

  3. Fink, D. et al. The role of DNA mismatch repair in platinum drug resistance. Cancer Res. 56, 4881–4886 (1996).

    CAS  Google Scholar 

  4. Krynetskaia, N.F. et al. Msh2 deficiency attenuates but does not abolish thiopurine hematopoietic toxicity in Msh2−/− mice. Mol. Pharmacol. 64, 456–465 (2003).

    Article  CAS  Google Scholar 

  5. Swann, P.F. et al. Role of postreplicative DNA mismatch repair in the cytotoxic action of thioguanine. Science 273, 1109–1111 (1996).

    Article  CAS  Google Scholar 

  6. Markowitz, S.D. & Bertagnolli, M.M. Molecular origins of cancer: molecular basis of colorectal cancer. N. Engl. J. Med. 361, 2449–2460 (2009).

    Article  CAS  Google Scholar 

  7. Lynch, H.T. et al. Hereditary ovarian carcinoma: heterogeneity, molecular genetics, pathology, and management. Mol. Oncol. 3, 97–137 (2009).

    Article  CAS  Google Scholar 

  8. Rowley, P.T. Inherited susceptibility to colorectal cancer. Annu. Rev. Med. 56, 539–554 (2005).

    Article  CAS  Google Scholar 

  9. Yuen, S.T. et al. Germline, somatic and epigenetic events underlying mismatch repair deficiency in colorectal and HNPCC-related cancers. Oncogene 21, 7585–7592 (2002).

    Article  CAS  Google Scholar 

  10. Ligtenberg, M.J. et al. Heritable somatic methylation and inactivation of MSH2 in families with Lynch syndrome due to deletion of the 3′ exons of TACSTD1. Nat. Genet. 41, 112–117 (2009).

    Article  CAS  Google Scholar 

  11. Matheson, E.C. & Hall, A.G. Expression of DNA mismatch repair proteins in acute lymphoblastic leukaemia and normal bone marrow. Adv. Exp. Med. Biol. 457, 579–583 (1999).

    Article  CAS  Google Scholar 

  12. Zhu, Y.M., Das-Gupta, E.P. & Russell, N.H. Microsatellite instability and p53 mutations are associated with abnormal expression of the MSH2 gene in adult acute leukemia. Blood 94, 733–740 (1999).

    CAS  Google Scholar 

  13. Marra, G. et al. Mismatch repair deficiency associated with overexpression of the MSH3 gene. Proc. Natl. Acad. Sci. USA 95, 8568–8573 (1998).

    Article  CAS  Google Scholar 

  14. Belloni, M., Uberti, D., Rizzini, C., Jiricny, J. & Memo, M. Induction of two DNA mismatch repair proteins, MSH2 and MSH6, in differentiated human neuroblastoma SH-SY5Y cells exposed to doxorubicin. J. Neurochem. 72, 974–979 (1999).

    Article  CAS  Google Scholar 

  15. Dosch, J., Christmann, M. & Kaina, B. Mismatch G-T binding activity and MSH2 expression is quantitatively related to sensitivity of cells to methylating agents. Carcinogenesis 19, 567–573 (1998).

    Article  CAS  Google Scholar 

  16. Hernandez-Pigeon, H., Laurent, G., Humbert, O., Salles, B. & Lautier, D. Degradation of mismatch repair hMutSα heterodimer by the ubiquitin-proteasome pathway. FEBS Lett. 562, 40–44 (2004).

    Article  CAS  Google Scholar 

  17. Valeri, N. et al. Modulation of mismatch repair and genomic stability by miR-155. Proc. Natl. Acad. Sci. USA 107, 6982–6987 (2010).

    Article  CAS  Google Scholar 

  18. Mullighan, C.G. et al. Genome-wide analysis of genetic alterations in acute lymphoblastic leukaemia. Nature 446, 758–764 (2007).

    Article  CAS  Google Scholar 

  19. Hernandez-Pigeon, H. et al. hMutSα is protected from ubiquitin-proteasome-dependent degradation by atypical protein kinase C ζ phosphorylation. J. Mol. Biol. 348, 63–74 (2005).

    Article  CAS  Google Scholar 

  20. Pui, C.H., Relling, M.V. & Downing, J.R. Acute lymphoblastic leukemia. N. Engl. J. Med. 350, 1535–1548 (2004).

    Article  CAS  Google Scholar 

  21. Karran, P. & Attard, N. Thiopurines in current medical practice: molecular mechanisms and contributions to therapy-related cancer. Nat. Rev. Cancer 8, 24–36 (2008).

    Article  CAS  Google Scholar 

  22. Gingras, A.C., Raught, B. & Sonenberg, N. Regulation of translation initiation by FRAP/mTOR. Genes Dev. 15, 807–826 (2001).

    Article  CAS  Google Scholar 

  23. Sontag, E., Sontag, J.M. & Garcia, A. Protein phosphatase 2A is a critical regulator of protein kinase C ζ signaling targeted by SV40 small t to promote cell growth and NF-κB activation. EMBO J. 16, 5662–5671 (1997).

    Article  CAS  Google Scholar 

  24. Chong-Kopera, H. et al. TSC1 stabilizes TSC2 by inhibiting the interaction between TSC2 and the HERC1 ubiquitin ligase. J. Biol. Chem. 281, 8313–8316 (2006).

    Article  CAS  Google Scholar 

  25. Brugarolas, J. et al. Regulation of mTOR function in response to hypoxia by REDD1 and the TSC1/TSC2 tumor suppressor complex. Genes Dev. 18, 2893–2904 (2004).

    Article  CAS  Google Scholar 

  26. Gao, X. & Pan, D. TSC1 and TSC2 tumor suppressors antagonize insulin signaling in cell growth. Genes Dev. 15, 1383–1392 (2001).

    Article  CAS  Google Scholar 

  27. Falasca, M. & Maffucci, T. Role of class II phosphoinositide 3-kinase in cell signaling. Biochem. Soc. Trans. 35, 211–214 (2007).

    Article  CAS  Google Scholar 

  28. de Wind, N., Dekker, M., van Rossum, A., van der Valk, M. & te Riele, H. Mouse models for hereditary nonpolyposis colorectal cancer. Cancer Res. 58, 248–255 (1998).

    CAS  Google Scholar 

  29. Bouffler, S.D., Hofland, N., Cox, R. & Fodde, R. Evidence for Msh2 haploinsufficiency in mice revealed by MNU-induced sister-chromatid exchange analysis. Br. J. Cancer 83, 1291–1294 (2000).

    Article  CAS  Google Scholar 

  30. DeWeese, T.L. et al. Mouse embryonic stem cells carrying one or two defective Msh2 alleles respond abnormally to oxidative stress inflicted by low-level radiation. Proc. Natl. Acad. Sci. USA 95, 11915–11920 (1998).

    Article  CAS  Google Scholar 

  31. Firestein, R. et al. CDK8 is a colorectal cancer oncogene that regulates β-catenin activity. Nature 455, 547–551 (2008).

    Article  CAS  Google Scholar 

  32. Paulsson, K. et al. Microdeletions are a general feature of adult and adolescent acute lymphoblastic leukemia: unexpected similarities with pediatric disease. Proc. Natl. Acad. Sci. USA 105, 6708–6713 (2008).

    Article  CAS  Google Scholar 

  33. Kuismanen, S.A. et al. Epigenetic phenotypes distinguish microsatellite-stable and -unstable colorectal cancers. Proc. Natl. Acad. Sci. USA 96, 12661–12666 (1999).

    Article  CAS  Google Scholar 

  34. Bellacosa, A. Functional interactions and signaling properties of mammalian DNA mismatch repair proteins. Cell Death Differ. 8, 1076–1092 (2001).

    Article  CAS  Google Scholar 

  35. Branch, P., Aquilina, G., Bignami, M. & Karran, P. Defective mismatch binding and a mutator phenotype in cells tolerant to DNA damage. Nature 362, 652–654 (1993).

    Article  CAS  Google Scholar 

  36. Papaefthymiou, M.A., Giaginis, C.T. & Theocharis, S.E. DNA repair alterations in common pediatric malignancies. Med. Sci. Monit. 14, RA8–RA15 (2008).

    CAS  Google Scholar 

Download references

Acknowledgements

We gratefully acknowledge the subjects and parents who participated in this study and the outstanding technical support of the Hartwell Center for Bioinformatics and Biotechnology at St. Jude Children's Research Hospital. We also thank Y. Wang, T. Brooks, J. Smith, W. Du, S. Mukatira, Y. Chu, M. Needham, P. Hargrove, G. Stocco and S. Paugh for their advice and technical support; J. Groff for preparation of the figures; K. Crews, N. Kornegay and M. Wilkinson for their research database expertise; J.C. Panetta for his modeling expertise; J. Jenkins for his immunohistochemistry expertise; T. Kunkel and A.B. Clark (National Institute of Environmental Health Sciences) for providing the E. coli strains, the wild-type and mutant M13mp2 phage and for their contributions to our MMR experiments; and J. Luis Rosa (Universitat de Barcelona) for providing us with antibodies to HERC1. We thank M. Kastan and D. Green for their critical review and advice. This work was supported in part by grant R37 CA36401 (W.E.E. and M.V.R.), NIH National Institute of General Medical Sciences Pharmacogenomics Research Network grant U01 GM92666 (M.V.R. and W.E.E.), CGM is a Pew Scholar and a St. Baldrick's scholar, and St. Jude is supported by a Cancer Center Support Grant CA 21765 from the National Cancer Institute and by the American Lebanese Syrian Associated Charities (ALSAC). H.G., S.C. and P.H. were funded by the Intramural Research Program of the National Institute of Diabetes and Digestive and Kidney Diseases of the NIH.

Author information

Authors and Affiliations

  1. Hematological Malignancies Program, St. Jude Children's Research Hospital, Memphis, Tennessee, USA

    Barthelemy Diouf, Qing Cheng, Natalia F Krynetskaia, Wenjian Yang, Meyling Cheok, Deqing Pei, Cheng Cheng, Evgeny Y Krynetskiy, William E Thierfelder, Charles G Mullighan, James R Downing, Ching-Hon Pui, Mary V Relling & William E Evans

  2. Department of Pharmaceutical Sciences, St. Jude Children's Research Hospital, Memphis, Tennessee, USA

    Barthelemy Diouf, Qing Cheng, Natalia F Krynetskaia, Wenjian Yang, Meyling Cheok, Evgeny Y Krynetskiy, William E Thierfelder, Mary V Relling & William E Evans

  3. Department of Biostatistics, St. Jude Children's Research Hospital, Memphis, Tennessee, USA

    Deqing Pei & Cheng Cheng

  4. Hartwell Center for Bioinformatics and Biotechnology, St. Jude Children's Research Hospital, Memphis, Tennessee, USA

    Yiping Fan

  5. Genetics and Biochemistry Branch, National Institute of Diabetes and Digestive and Kidney Diseases, US National Institutes of Health (NIH), Bethesda, Maryland, USA

    Hui Geng, Siying Chen & Peggy Hsieh

  6. Department of Pathology, St. Jude Children's Research Hospital, Memphis, Tennessee, USA

    Charles G Mullighan, James R Downing & Ching-Hon Pui

  7. Department of Oncology, St. Jude Children's Research Hospital, Memphis, Tennessee, USA

    Ching-Hon Pui

Authors
  1. Barthelemy Diouf
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Qing Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Natalia F Krynetskaia
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wenjian Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Meyling Cheok
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Deqing Pei
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yiping Fan
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Cheng Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Evgeny Y Krynetskiy
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Hui Geng
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Siying Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. William E Thierfelder
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Charles G Mullighan
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. James R Downing
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Peggy Hsieh
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Ching-Hon Pui
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. Mary V Relling
    View author publications

    You can also search for this author in PubMed Google Scholar

  18. William E Evans
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

W.E.E. designed and supervised experiments and their analyses and wrote the manuscript with B.D. B.D., Q.C., N.F.K., M.C., E.Y.K., H.G., S.C., P.H., W.E.T. and C.G.M. performed experiments and participated in their analyses. J.R.D., C.G.M. and M.V.R. directed experiments and contributed to the genomic analyses. D.P., Y.F. and C.C. performed the statistical analyses. W.Y. led the genomic analyses in collaboration with other authors. C.-H.P. led the clinical trials and provided the ALL samples. All authors discussed the results and commented on the manuscript.

Corresponding author

Correspondence to William E Evans.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–10, Supplementary Tables 1–8 and Supplementary Methods (PDF 2015 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Diouf, B., Cheng, Q., Krynetskaia, N. et al. Somatic deletions of genes regulating MSH2 protein stability cause DNA mismatch repair deficiency and drug resistance in human leukemia cells. Nat Med 17, 1298–1303 (2011). https://doi.org/10.1038/nm.2430

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm.2430

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing