SHP2<sup>E76K</sup> mutant promotes lung tumorigenesis in transgenic mice

Valentina Schneeberger

Noreen Luetteke

Yuan Ren

Hartmut Berns

Jiandong Chen+2 authors

Supplementary Material

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^{Department of Molecular Oncology, H. Lee Moffitt Cancer Center and Research Institute,}

^{Division of Cell Biology, Microbiology, and Molecular Biology, University of South Florida,}

^{Small Animal Modeling and Imaging Core, and}

^{Department of Thoracic Oncology, H. Lee Moffitt Cancer Center and Research Institute,}

^{Department of Oncologic Sciences, University of South Florida College of Medicine , and}

^{Department of Anatomic Pathology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612, USA}

*To whom correspondence should be addressed. Department of Molecular Oncology, SRB-3, H. Lee Moffitt Cancer Center and Research Institute, 12902 Magnolia Drive, Tampa, FL 33612, USA. Tel: +1 813 745 6713; Fax: +1 813 745 3829; Email: gro.ttiffom@uw.yrrej

Received 2013 Sep 4; Revised 2013 Dec 20; Accepted 2014 Jan 17.

Abstract

Lung cancer is a major disease carrying heterogeneous molecular lesions and many of them remain to be analyzed functionally in vivo. Gain-of-function (GOF) SHP2 (PTPN11) mutations have been found in various types of human cancer, including lung cancer. However, the role of activating SHP2 mutants in lung cancer has not been established. We generated transgenic mice containing a doxycycline (Dox)-inducible activating SHP2 mutant (tetO-SHP2^{) and analyzed the role of SHP2}^{in lung tumorigenesis in the Clara cell secretory protein (CCSP)-reverse tetracycline transactivator (rtTA)/tetO-SHP2}^{bitransgenic mice. SHP2}^{activated Erk1/Erk2 (Erk1/2) and Src, and upregulated c-Myc and Mdm2 in the lungs of bitransgenic mice. Atypical adenomatous hyperplasia and small adenomas were observed in CCSP-rtTA/tetO-SHP2}^{bitransgenic mice induced with Dox for 2–6 months and progressed to larger adenoma and adenocarcinoma by 9 months. Dox withdrawal from bitransgenic mice bearing magnetic resonance imaging-detectable lung tumors resulted in tumor regression. These results show that the activating SHP2 mutant promotes lung tumorigenesis and that the SHP2 mutant is required for tumor maintenance in this mouse model of non-small cell lung cancer. SHP2}^{was associated with Gab1 in the lung of transgenic mice. Elevated pGab1 was observed in the lung of Dox-induced CCSP-rtTA/tetO-SHP2}^{mice and in cell lines expressing SHP2}^{, indicating that the activating SHP2 mutant autoregulates tyrosine phosphorylation of its own docking protein. Gab1 tyrosine phosphorylation is sensitive to inhibition by the Src inhibitor dasatinib in GOF SHP2-mutant-expressing cells, suggesting that Src family kinases are involved in SHP2 mutant-induced Gab1 tyrosine phosphorylation.}

Abstract

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Acknowledgements

We thank J.A.Whitset for the CCSP-rtTA transgenic mice, D.C.Radisky and A.P.Fields for advice and assistance, K.Politi and G.Felsenfeld for reagents, and E.Ruiz, A.Lopez and the Moffitt Animal, Tissue, and Microscopy Core staffs for assistance.

Conflict of Interest Statement: None declared.

Acknowledgements

Glossary

Abbreviations:

CCSP	Clara cell secretory protein
Dox	doxycyline
EGFR	epidermal growth factor receptor
GOF	gain-of-function
H&E	hematoxylin and eosin
mRNA	messenger RNA
MRI	magnetic resonance imaging
NSCLC	non-small cell lung cancer
PTK	protein tyrosine kinase
PTP	protein tyrosine phosphatase
RMCE	recombinase-mediated cassette exchange
rtTA	reverse tetracycline transactivator
RT–PCR	reverse transcription–PCR
SFKs	Src family kinases
siRNA	small interfering RNA.

Glossary

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