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Suppression of lung adenocarcinoma progression by Nkx2-1

Abstract

Despite the high prevalence and poor outcome of patients with metastatic lung cancer the mechanisms of tumour progression and metastasis remain largely uncharacterized. Here we modelled human lung adenocarcinoma, which frequently harbours activating point mutations in KRAS1 and inactivation of the p53 pathway2, using conditional alleles in mice3,4,5. Lentiviral-mediated somatic activation of oncogenic Kras and deletion of p53 in the lung epithelial cells of KrasLSL-G12D/+;p53flox/flox mice initiates lung adenocarcinoma development4. Although tumours are initiated synchronously by defined genetic alterations, only a subset becomes malignant, indicating that disease progression requires additional alterations. Identification of the lentiviral integration sites allowed us to distinguish metastatic from non-metastatic tumours and determine the gene expression alterations that distinguish these tumour types. Cross-species analysis identified the NK2-related homeobox transcription factor Nkx2-1 (also called Ttf-1 or Titf1) as a candidate suppressor of malignant progression. In this mouse model, Nkx2-1 negativity is pathognomonic of high-grade poorly differentiated tumours. Gain- and loss-of-function experiments in cells derived from metastatic and non-metastatic tumours demonstrated that Nkx2-1 controls tumour differentiation and limits metastatic potential in vivo. Interrogation of Nkx2-1-regulated genes, analysis of tumours at defined developmental stages, and functional complementation experiments indicate that Nkx2-1 constrains tumours in part by repressing the embryonically restricted chromatin regulator Hmga2. Whereas focal amplification of NKX2-1 in a fraction of human lung adenocarcinomas has focused attention on its oncogenic function6,7,8,9, our data specifically link Nkx2-1 downregulation to loss of differentiation, enhanced tumour seeding ability and increased metastatic proclivity. Thus, the oncogenic and suppressive functions of Nkx2-1 in the same tumour type substantiate its role as a dual function lineage factor.

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Figure 1: A lentiviral vector-induced mouse model of lung adenocarcinoma identifies gene expression alterations during tumour progression.
Figure 2: Reduced Nkx2-1 expression in advanced lung adenocarcinoma correlates with a less differentiated state.
Figure 3: Nkx2-1 controls lung adenocarcinoma differentiation and restricts metastatic ability.
Figure 4: Nkx2-1 regulates the expression of Hmga2 in advanced lung adenocarcinoma.

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Acknowledgements

We thank A. Dooley, N. Dimitrova, T. Oliver and M. Ebert for providing reagents; M. Luo (Biology/Koch Institute BioMicro Core) for array processing; M. Kumar for experimental assistance; T. Staton, D. McFadden, A. Shaw and the entire T.J. laboratory for comments. M.M.W. was a Merck Fellow of the Damon Runyon Cancer Research Foundation and was funded by a Genentech Postdoctoral Fellowship. R.G.W.V. is supported by a Fellowship from the Dutch Cancer Society KWF. E.L.S. is supported by a training grant (T32-HL007627). D.Y.C. is supported by an Alfred P. Sloan Foundation Research Fellowship. D.M.F. is a Leukemia and Lymphoma Postdoctoral Fellow. This work was supported by National Institutes of Health grants U01-CA84306 (to T.J.) and K99-CA151968 (to M.M.W.), the Howard Hughes Medical Institute, the Ludwig Center for Molecular Oncology at MIT, and in part by the Cancer Center Support (core) grant P30-CA14051 from the National Cancer Institute. T.J. is the David H. Koch Professor of Biology and a Daniel K. Ludwig Scholar.

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Contributions

M.M.W. and T.J. designed the study; M.M.W., T.L.D., C.K.-K. and E.L.S. performed experiments; R.G.W.V., M.M.W., C.A.W., D.D.H., S.H. and D.Y.C. conducted bioinformatic analyses; E.L.S. and R.T.B. provided pathology assistance; S.Y. and D.C. provided technical assistance; M.J.D. provided reagents; D.M.F. and M.M. gave conceptual advice; M.M.W. and T.J. wrote the paper with comments from all authors.

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Correspondence to Tyler Jacks.

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The authors declare no competing financial interests.

Additional information

Gene expression data was deposited in Gene Expression Omnibus (GSE26874).

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Supplementary Figures

This file contains Supplementary Figures 1-13 with legends. (PDF 5817 kb)

Supplementary Table 1

This table contains the gene expression data for the TnonMet and advanced tumor-derived (TMet/Met) samples. Cell lines are listed and gene expression is shown. (XLS 10790 kb)

Supplementary Table 2

This table contains the Nkx2-1 regulated genes. Gene expression data for the TnonMet-control and TnonMet-shNkx2-1 cells is shown (XLS 6681 kb)

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Winslow, M., Dayton, T., Verhaak, R. et al. Suppression of lung adenocarcinoma progression by Nkx2-1. Nature 473, 101–104 (2011). https://doi.org/10.1038/nature09881

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