Oncotarget

Research Papers: Chromosome:

A novel gammaretroviral shuttle vector insertional mutagenesis screen identifies SHARPIN as a breast cancer metastasis gene and prognostic biomarker

Victor M. Bii, Dustin T. Rae, Grant D. Trobridge _

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Oncotarget. 2015; 6:39507-39520. https://doi.org/10.18632/oncotarget.6232

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Abstract

Victor M. Bii1, Dustin T. Rae1 and Grant D. Trobridge1,2

1 Washington State University College of Pharmacy, WSU Spokane, Spokane, WA, USA

2 School of Molecular Biosciences, Washington State University, Pullman, Washington, USA

Correspondence to:

Grant D. Trobridge, email:

Keywords: insertional mutagenesis screen, breast cancer, gammaretroviral vector (γRV), metastasis, prognostic biomarker, Chromosome Section

Received: October 02, 2015 Accepted: October 14, 2015 Published: October 25, 2015

Abstract

Breast cancer (BC) is the second leading cause of malignancy among U.S. women. Metastasis results in a poor prognosis and increased mortality, but the molecular mechanisms by which metastatic tumors occur are not well understood. Identifying the genes that drive the metastatic process could provide targets for improved therapy and biomarkers to improve BC patient outcomes. Using a forward mutagenesis screen, BC cells mutagenized with a replication-incompetent gammaretroviral vector (γRV) were xenotransplanted into the mammary fat pad of immunodeficient mice. In this approach the vector provirus dysregulates nearby genes, providing a selective advantage to transduced cells to form metastases. Metastatic tumors were analyzed for proviral integration sites to identify nearby candidate metastasis genes. The γRV has a transgene cassette that allows for rescue in bacteria and rapid identification of vector integration sites. Using this approach, we identified the previously described metastasis gene WWTR1 (TAZ), and three other novel candidate metastasis genes including SHARPIN. SHARPIN was independently validated in vivo as a BC metastasis gene. Analysis of patient data showed that SHARPIN expression predicts metastasis-free survival after adjuvant therapy. Our approach has broad potential to identify genes involved in oncogenic processes for BC and other cancers. We show here it can identify both known (WWTR1) and novel (SHARPIN) BC metastasis genes.

Author Information

Victor M. Bii

Dustin T. Rae

Grant D. Trobridge
Primary Contact  _


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