Ablation of the Kinase NDR1 Predisposes Mice to the Development of T Cell Lymphoma
Lymphoma from Compromised Apoptosis
The four members of the nuclear Dbf2–related (NDR) family of serine and threonine protein kinases act in a number of cellular processes, including proliferation, cytokinesis, and apoptosis. Two family members, LATS1 and LATS2, act as tumor suppressor proteins in flies and mice. Of the other two, NDR1 is abundant in organs of the immune system, whereas NDR2 is predominantly found in the gastrointestinal tract. Cornils et al. found that mouse lymphocytes deficient in NDR1 compensated for this loss by increasing the abundance of NDR2 protein in a posttranscriptional manner. Blockade of this compensatory mechanism resulted in increased resistance of cells to various extrinsic and intrinsic proapoptotic stimuli. Aged NDR1+/− and NDR1−/− mice were more susceptible to the development of T cell lymphoma than were their wild-type counterparts, and this was associated with a loss in total NDR protein. Correlating with this finding, the abundance of NDR proteins was less in samples of human T cell lymphomas than in normal T cells, suggesting that NDR1 acts as a tumor suppressor protein.
Abstract
Defective apoptosis contributes to the development of various human malignancies. The kinases nuclear Dbf2–related 1 (NDR1) and NDR2 mediate apoptosis downstream of the tumor suppressor proteins RASSF1A (Ras association domain family member 1A) and MST1 (mammalian Ste20-like kinase 1). To further analyze the role of NDR1 in apoptosis, we generated NDR1-deficient mice. Although NDR1 is activated by both intrinsic and extrinsic proapoptotic stimuli, which indicates a role for NDR1 in regulating apoptosis, NDR1-deficient T cells underwent apoptosis in a manner similar to that of wild-type cells in response to different proapoptotic stimuli. Analysis of the abundances of NDR1 and NDR2 proteins revealed that loss of NDR1 was functionally compensated for by an increase in the abundance of NDR2 protein. Despite this compensation, NDR1−/− and NDR1+/− mice were more prone to the development of T cell lymphomas than were wild-type mice. Tumor development in mice and humans was accompanied by a decrease in the overall amounts of NDR proteins in T cell lymphoma samples. Thus, reduction in the abundance of NDR1 triggered a decrease in the total amount of both isoforms. Together, our data suggest that a reduction in the abundances of the NDR proteins results in defective responses to proapoptotic stimuli, thereby facilitating the development of tumors.
Get full access to this article
View all available purchase options and get full access to this article.
Already a Subscriber?Sign In
Supplementary Material
Summary
Fig. S1. Generation of NDR1-deficient mice.
Fig. S2. Mature T cells from NDR1+/− and NDR1−/− mice show slightly increased resistance to apoptosis.
Fig. S3. Activation of NDR is unchanged in NDR1-deficient thymocytes in response to induction of apoptosis by dexamethasone or antibody against Fas together with cycloheximide.
Fig. S4. NDR1 and NDR2 show distinct patterns in mice.
Fig. S5. The expression of NDR2 and the abundance of NDR2 protein remain unchanged upon loss of NDR1.
Fig. S6. Reduction of the abundance of NDR2 in NDR1-deficient MEFs results in increased resistance to apoptosis.
Fig. S7. Increased development of MPDs in NDR1+/− and NDR1−/− mice after ENU treatment.
Fig. S8. An increase in the abundance of Pou2af1 correlates with a low abundance of NDR in tumors.
Fig. S9. Loss of NDR1 results in minor defects in the proliferation of mature T cells after stimulation of the TCR.
Fig. S10. Cells from NDR-high and NDR-low tumors show no consistent differences in Ki67 staining.
Fig. S11. siRNA against MEF2c does not rescue apoptosis defects in cells depleted of NDR1 and NDR2.
Fig. S12. Characterization of a murine antibody against NDR1.
Fig. S13. Validation of siRNA against MEF2c and overexpression of E47 in transfected cells.
Fig. S14. Testing of different shNDR2 constructs.
Table S1. Aged mice analyzed for tumor development.
Table S2. Overview of tumors identified in ENU-treated mice.
Table S3. Classification of human T cell lymphoma samples.
Table S4. Classification of tumors according to the abundances of NDR1 and NDR2.
Table S5. Genes whose expression was significantly altered in NDR-low tumors.
Resources
File (3_ra47_sm.pdf)
References and Notes
1
Taylor R. C., Cullen S. P., Martin S. J., Apoptosis: Controlled demolition at the cellular level. Nat. Rev. Mol. Cell Biol. 9, 231–241 (2008).
2
Kurokawa M., Kornbluth S., Caspases and kinases in a death grip. Cell 138, 838–854 (2009).
3
Radu M., Chernoff J., The DeMSTification of mammalian Ste20 kinases. Curr. Biol. 19, R421–R425 (2009).
4
Vichalkovski A., Gresko E., Cornils H., Hergovich A., Schmitz D., Hemmings B. A., NDR kinase is activated by RASSF1A/MST1 in response to Fas receptor stimulation and promotes apoptosis. Curr. Biol. 18, 1889–1895 (2008).
5
Chan E. H., Nousiainen M., Chalamalasetty R. B., Schäfer A., Nigg E. A., Sillje H. H., The Ste20-like kinase Mst2 activates the human large tumor suppressor kinase Lats1. Oncogene 24, 2076–2086 (2005).
6
Hergovich A., Stegert M. R., Schmitz D., Hemmings B. A., NDR kinases regulate essential cell processes from yeast to humans. Nat. Rev. Mol. Cell Biol. 7, 253–264 (2006).
7
Pan D., Hippo signaling in organ size control. Genes Dev. 21, 886–897 (2007).
8
Zeng Q., Hong W., The emerging role of the Hippo pathway in cell contact inhibition, organ size control, and cancer development in mammals. Cancer Cell 13, 188–192 (2008).
9
Chiba S., Ikeda M., Katsunuma K., Ohashi K., Mizuno K., MST2- and Furry-mediated activation of NDR1 kinase is critical for precise alignment of mitotic chromosomes. Curr. Biol. 19, 675–681 (2009).
10
Hergovich A., Lamla S., Nigg E. A., Hemmings B. A., Centrosome-associated NDR kinase regulates centrosome duplication. Mol. Cell 25, 625–634 (2007).
11
Stegert M. R., Tamaskovic R., Bichsel S. J., Hergovich A., Hemmings B. A., Regulation of NDR2 protein kinase by multi-site phosphorylation and the S100B calcium-binding protein. J. Biol. Chem. 279, 23806–23812 (2004).
12
Geng W., He B., Wang M., Adler P. N., The tricornered gene, which is required for the integrity of epidermal cell extensions, encodes the Drosophila nuclear DBF2-related kinase. Genetics 156, 1817–1828 (2000).
13
Fenske T. S., McMahon C., Edwin D., Jarvis J. C., Cheverud J. M., Minn M., Mathews V., Bogue M. A., Province M. A., McLeod H. L., Graubert T. A., Identification of candidate alkylator-induced cancer susceptibility genes by whole genome scanning in mice. Cancer Res. 66, 5029–5038 (2006).
14
Joslin J. M., Fernald A. A., Tennant T. R., Davis E. M., Kogan S. C., Anastasi J., Crispino J. D., Le Beau M. M., Haploinsufficiency of EGR1, a candidate gene in the del(5q), leads to the development of myeloid disorders. Blood 110, 719–726 (2007).
15
Geisen C., Karsunky H., Yucel R., Moroy T., Loss of p27Kip1 cooperates with cyclin E in T-cell lymphomagenesis. Oncogene 22, 1724–1729 (2003).
16
Kang-Decker N., Tong C., Boussouar F., Baker D. J., Xu W., Leontovich A. A., Taylor W. R., Brindle P. K., van Deursen J. M., Loss of CBP causes T cell lymphomagenesis in synergy with p27Kip1 insufficiency. Cancer Cell 5, 177–189 (2004).
17
Heckman C. A., Duan H., Garcia P. B., Boxer L. M., Oct transcription factors mediate t(14;18) lymphoma cell survival by directly regulating bcl-2 expression. Oncogene 25, 888–898 (2006).
18
Miyazaki T., Hirokami Y., Matsuhashi N., Takatsuka H., Naito M., Increased susceptibility of thymocytes to apoptosis in mice lacking AIM, a novel murine macrophage-derived soluble factor belonging to the scavenger receptor cysteine-rich domain superfamily. J. Exp. Med. 189, 413–422 (1999).
19
Nagel S., Meyer C., Quentmeier H., Kaufmann M., Drexler H. G., MacLeod R. A., MEF2C is activated by multiple mechanisms in a subset of T-acute lymphoblastic leukemia cell lines. Leukemia 22, 600–607 (2008).
20
Burgess D. J., Doles J., Zender L., Xue W., Ma B., McCombie W. R., Hannon G. J., Lowe S. W., Hemann M. T., Topoisomerase levels determine chemotherapy response in vitro and in vivo. Proc. Natl. Acad. Sci. U.S.A. 105, 9053–9058 (2008).
21
Kulkarni M. S., Daggett J. L., Bender T. P., Kuehl W. M., Bergsagel P. L., Williams M. E., Frequent inactivation of the cyclin-dependent kinase inhibitor p18 by homozygous deletion in multiple myeloma cell lines: Ectopic p18 expression inhibits growth and induces apoptosis. Leukemia 16, 127–134 (2002).
22
Mendelsohn A. R., Hamer J. D., Wang Z. B., Brent R., Cyclin D3 activates Caspase 2, connecting cell proliferation with cell death. Proc. Natl. Acad. Sci. U.S.A. 99, 6871–6876 (2002).
23
Tsuchihara K., Lapin V., Bakal C., Okada H., Brown L., Hirota-Tsuchihara M., Zaugg K., Ho A., Itie-Youten A., Harris-Brandts M., Rottapel R., Richardson C. D., Benchimol S., Mak T. W., Ckap2 regulates aneuploidy, cell cycling, and cell death in a p53-dependent manner. Cancer Res. 65, 6685–6691 (2005).
24
Kee B. L., E and ID proteins branch out. Nat. Rev. Immunol. 9, 175–184 (2009).
25
Bain G., Engel I., Robanus Maandag E. C., te Riele H. P., Voland J. R., Sharp L. L., Chun J., Huey B., Pinkel D., Murre C., E2A deficiency leads to abnormalities in αβ T-cell development and to rapid development of T-cell lymphomas. Mol. Cell. Biol. 17, 4782–4791 (1997).
26
O’Neil J., Shank J., Cusson N., Murre C., Kelliher M., TAL1/SCL induces leukemia by inhibiting the transcriptional activity of E47/HEB. Cancer Cell 5, 587–596 (2004).
27
Yan W., Young A. Z., Soares V. C., Kelley R., Benezra R., Zhuang Y., High incidence of T-cell tumors in E2A-null mice and E2A/Id1 double-knockout mice. Mol. Cell. Biol. 17, 7317–7327 (1997).
28
Santarosa M., Ashworth A., Haploinsufficiency for tumour suppressor genes: When you don’t need to go all the way. Biochim. Biophys. Acta 1654, 105–122 (2004).
29
Raval A., Tanner S. M., Byrd J. C., Angerman E. B., Perko J. D., Chen S. S., Hackanson B., Grever M. R., Lucas D. M., Matkovic J. J., Lin T. S., Kipps T. J., Murray F., Weisenburger D., Sanger W., Lynch J., Watson P., Jansen M., Yoshinaga Y., Rosenquist R., de Jong P. J., Coggill P., Beck S., Lynch H., de la Chapelle A., Plass C., Downregulation of death-associated protein kinase 1 (DAPK1) in chronic lymphocytic leukemia. Cell 129, 879–890 (2007).
30
Valle L., Serena-Acedo T., Liyanarachchi S., Hampel H., Comeras I., Li Z., Zeng Q., Zhang H. T., Pennison M. J., Sadim M., Pasche B., Tanner S. M., de la Chapelle A., Germline allele-specific expression of TGFBR1 confers an increased risk of colorectal cancer. Science 321, 1361–1365 (2008).
31
Yan H., Dobbie Z., Gruber S. B., Markowitz S., Romans K., Giardiello F. M., Kinzler K. W., Vogelstein B., Small changes in expression affect predisposition to tumorigenesis. Nat. Genet. 30, 25–26 (2002).
32
Hanahan D., Weinberg R. A., The hallmarks of cancer. Cell 100, 57–70 (2000).
33
Hakem R., de la Pompa J. L., Sirard C., Mo R., Woo M., Hakem A., Wakeham A., Potter J., Reitmair A., Billia F., Firpo E., Hui C. C., Roberts J., Rossant J., Mak T. W., The tumor suppressor gene Brca1 is required for embryonic cellular proliferation in the mouse. Cell 85, 1009–1023 (1996).
34
Mack F. A., Patel J. H., Biju M. P., Haase V. H., Simon M. C., Decreased growth of Vhl−/− fibrosarcomas is associated with elevated levels of cyclin kinase inhibitors p21 and p27. Mol. Cell. Biol. 25, 4565–4578 (2005).
35
Yang X., Li C., Xu X., Deng C., The tumor suppressor SMAD4/DPC4 is essential for epiblast proliferation and mesoderm induction in mice. Proc. Natl. Acad. Sci. U.S.A. 95, 3667–3672 (1998).
36
Engel I., Murre C., Ectopic expression of E47 or E12 promotes the death of E2A-deficient lymphomas. Proc. Natl. Acad. Sci. U.S.A. 96, 996–1001 (1999).
37
Harima Y., Harima K., Sawada S., Tanaka Y., Arita S., Ohnishi T., Loss of heterozygosity on chromosome 6p21.2 as a potential marker for recurrence after radiotherapy of human cervical cancer. Clin. Cancer Res. 6, 1079–1085 (2000).
38
Lukeis R., Irving L., Garson M., Hasthorpe S., Cytogenetics of non-small cell lung cancer: Analysis of consistent non-random abnormalities. Genes Chromosomes Cancer 2, 116–124 (1990).
39
Morita R., Ishikawa J., Tsutsumi M., Hikiji K., Tsukada Y., Kamidono S., Maeda S., Nakamura Y., Allelotype of renal cell carcinoma. Cancer Res. 51, 820–823 (1991).
40
Nagai H., Kinoshita T., Suzuki H., Hatano S., Murate T., Saito H., Identification and mapping of novel tumor suppressor loci on 6p in diffuse large B-cell non-Hodgkin’s lymphoma. Genes Chromosomes Cancer 25, 277–283 (1999).
41
Sato T., Saito H., Morita R., Koi S., Lee J. H., Nakamura Y., Allelotype of human ovarian cancer. Cancer Res. 51, 5118–5122 (1991).
42
Vogelstein B., Fearon E. R., Kern S. E., Hamilton S. R., Preisinger A. C., Nakamura Y., White R., Allelotype of colorectal carcinomas. Science 244, 207–211 (1989).
43
Agostinelli C., Piccaluga P. P., Went P., Rossi M., Gazzola A., Righi S., Sista T., Campidelli C., Zinzani P. L., Falini B., Pileri S. A., Peripheral T cell lymphoma, not otherwise specified: The stuff of genes, dreams and therapies. J. Clin. Pathol. 61, 1160–1167 (2008).
44
Tamaskovic R., Bichsel S. J., Rogniaux H., Stegert M. R., Hemmings B. A., Mechanism of Ca2+-mediated regulation of NDR protein kinase through autophosphorylation and phosphorylation by an upstream kinase. J. Biol. Chem. 278, 6710–6718 (2003).
45
Abbreviations for the amino acids are as follows: D, Asp; E, Glu; H, His; I, Ile; K, Lys; L, Leu; N, Asn; P, Pro; S, Ser; T, Thr; V, Val; and Y, Tyr.
46
Bordon A., Bosco N., Du Roure C., Bartholdy B., Kohler H., Matthias G., Rolink A. G., Matthias P., Enforced expression of the transcriptional coactivator OBF1 impairs B cell differentiation at the earliest stage of development. PLoS One 3, e4007 (2008).
47
Hergovich A., Bichsel S. J., Hemmings B. A., Human NDR kinases are rapidly activated by MOB proteins through recruitment to the plasma membrane and phosphorylation. Mol. Cell. Biol. 25, 8259–8272 (2005).
48
Van Parijs L., Refaeli Y., Lord J. D., Nelson B. H., Abbas A. K., Baltimore D., Uncoupling IL-2 signals that regulate T cell proliferation, survival, and Fas-mediated activation-induced cell death. Immunity 11, 281–288 (1999).
49
Senoo M., Manis J. P., Alt F. W., McKeon F., p63 and p73 are not required for the development and p53-dependent apoptosis of T cells. Cancer Cell 6, 85–89 (2004).
50
van de Wetering M., Oving I., Muncan V., Pon Fong M. T., Brantjes H., van Leenen D., Holstege F. C., Brummelkamp T. R., Agami R., Clevers H., Specific inhibition of gene expression using a stably integrated, inducible small-interfering-RNA vector. EMBO Rep. 4, 609–615 (2003).
Information & Authors
Information
Published In
Science Signaling
Volume 3 | Issue 126
June 2010
June 2010
Copyright
Copyright © 2010, American Association for the Advancement of Science.
Submission history
Received: 6 October 2009
Accepted: 27 May 2010
Acknowledgments
Acknowledgments: We thank L. Quintanilla-Fend (GSF, Munich, Germany) for help with the antibodies used for immunohistochemistry; A. Wodnar-Filipowicz (University of Basel, Basel, Switzerland) for providing purified human T cells; G. Holländer (University of Basel) and A. Cornils (Friedrich Miescher Institute, Basel, Switzerland) for discussion; and P. Morin Jr. (University of Moncton, Moncton, New Brunswick, Canada) for comments on the manuscript. Funding: This work was supported by the Novartis Research Foundation, the Swiss Cancer League, and the Boehringer Ingelheim Fonds. Author contributions: H.C. designed and performed the research, analyzed the data, and wrote the manuscript; M.R.S. generated NDR1-targeted mice; A.H. designed, cloned, and tested shRNA against mNDR2 and critically reviewed the manuscript; D.H. helped with mouse experiments; D.S. designed and tested quantitative PCR assays for detecting murine NDR2 mRNA; S.D. analyzed tissue samples; and B.A.H. initiated this project and assisted with research design and manuscript preparation. Competing interests: H.C. and B.A.H. have a patent application, PCT application WO2008/065391, related to this work. Accession numbers: Microarray data have been stored in the GEO database under accession number GSE21902.
Authors
Metrics & Citations
Metrics
Article Usage
Altmetrics
Citations
Export citation
Select the format you want to export the citation of this publication.
Cited by
- The “STK38-XPO1 axis”: its general relevance and mechanistic underpinnings remain to be further characterized, Cellular and Molecular Life Sciences, 78, 9, (4451-4452), (2021).https://doi.org/10.1007/s00018-021-03778-x
- hMOB2 deficiency impairs homologous recombination-mediated DNA repair and sensitises cancer cells to PARP inhibitors, Cellular Signalling, 87, (110106), (2021).https://doi.org/10.1016/j.cellsig.2021.110106
- tRNA overexpression rescues peripheral neuropathy caused by mutations in tRNA synthetase, Science, 373, 6559, (1161-1166), (2021)./doi/10.1126/science.abb3356
- Nuclear Dbf2-related Kinase 1 functions as tumor suppressor in glioblastoma by phosphorylation of Yes-associated protein, Chinese Medical Journal, 134, 17, (2054-2065), (2021).https://doi.org/10.1097/CM9.0000000000001653
- The kinases NDR1/2 act downstream of the Hippo homolog MST1 to mediate both egress of thymocytes from the thymus and lymphocyte motility, Science Signaling, 8, 397, (ra100-ra100), (2021)./doi/10.1126/scisignal.aab2425
- The STK38–XPO1 axis, a new actor in physiology and cancer, Cellular and Molecular Life Sciences, 78, 5, (1943-1955), (2020).https://doi.org/10.1007/s00018-020-03690-w
- STK38 promotes ATM activation by acting as a reader of histone H4 ufmylation, Science Advances, 6, 23, (2020)./doi/10.1126/sciadv.aax8214
- Furry protein suppresses nuclear localization of yes-associated protein (YAP) by activating NDR kinase and binding to YAP, Journal of Biological Chemistry, 295, 10, (3017-3028), (2020).https://doi.org/10.1074/jbc.RA119.010783
- The Hippo Pathway and Viral Infections, Frontiers in Microbiology, 10, (2020).https://doi.org/10.3389/fmicb.2019.03033
- The Emerging Roles of NDR1/2 in Infection and Inflammation, Frontiers in Immunology, 11, (2020).https://doi.org/10.3389/fimmu.2020.00534
Loading...
View Options
Get Access
Log in to view the full text
AAAS login provides access to Science for AAAS Members, and access to other journals in the Science family to users who have purchased individual subscriptions.
- Become a AAAS Member
- Activate your AAAS ID
- Purchase Access to Other Journals in the Science Family
- Account Help
Log in via OpenAthens.
Log in via Shibboleth.
More options
Register for free to read this article
As a service to the community, this article is available for free. Login or register for free to read this article.
View options
PDF format
Download this article as a PDF file
Download PDF