Abstract
Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal human cancers, with 5-year patient survival rates of <5%. Activating mutations in KRAS are the predominant oncogenic drivers of PDAC but are accompanied by additional lower frequency genetic alterations. Our group previously identified the guanine nucleotide exchange factor ARHGEF10 in a genomic screen for genes with copy number alterations that may synergize with oncogenic KRAS to promote PDAC carcinogenesis. In the present study we show that ARHGEF10 possesses putative tumor suppressor function in PDAC. ARHGEF10 expression is reduced in over 70% of PDAC cell lines, and copy number loss is documented in more than 30% of PDAC patient-derived xenografts. Loss of ARHGEF10 expression enhanced subcutaneous tumor growth in mouse models, while its exogenous expression greatly impaired tumorigenesis. Loss of ARHGEF10 expression also increased in vitro proliferation, invasion, and motility of PDAC cell lines, and enhanced their metastatic spread in orthotopic mouse models. Treatment of ARHGEF10-depleted cells with the inhibitor dasatinib reduced levels of phospho Src kinase and attenuated motility and invasion in vitro. Together, our data indicate that ARHGEF10 may function as a tumor suppressor in PDAC.
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References
Siegel RL, Miller KD, Jemal A. Cancer statistics. CA Cancer J Clin. 2016;66:7–30.
Rahib L, Smith BD, Aizenberg R, Rosenzweig AB, Fleshman JM, Matrisian LM. Projecting cancer incidence and deaths to 2030: the unexpected burden of thyroid, liver, and pancreas cancers in the United States. Cancer Res. 2014;74:2913–21.
Ryan DP, Hong TS, Bardeesy N. Pancreatic adenocarcinoma. N Engl J Med. 2014;371:2140–1.
Jones S, Zhang X, Parsons DW, Lin JC, Leary RJ, Angenendt P, et al. Core signaling pathways in human pancreatic cancers revealed by global genomic analyses. Science. 2008;321:1801–6.
Biankin AV, Waddell N, Kassahn KS, Gingras MC, Muthuswamy LB, Johns AL, et al. Pancreatic cancer genomes reveal aberrations in axon guidance pathway genes. Nature. 2012;491:399–405.
Waddell N, Pajic M, Patch AM, Chang DK, Kassahn KS, Bailey P, et al. Whole genomes redefine the mutational landscape of pancreatic cancer. Nature. 2015;518:495–501.
Radulovich N, Leung L, Ibrahimov E, Navab R, Sakashita S, Zhu CQ, et al. Coiled-coil domain containing 68 (CCDC68) demonstrates a tumor-suppressive role in pancreatic ductal adenocarcinoma. Oncogene. 2015;34:4238–47.
Mohl M, Winkler S, Wieland T, Lutz S. Gef10 - the third member of a Rho-specific guanine nucleotide exchange factor subfamily with unusual protein architecture. Naunyn Schmiede Arch Pharm. 2006;373:333–41.
Aoki T, Ueda S, Kataoka T, Satoh T. Regulation of mitotic spindle formation by the RhoA guanine nucleotide exchange factor ARHGEF10. BMC Cell Biol. 2009;10:56.
Bashyam MD, Bair R, Kim YH, Wang P, Hernandez-Boussard T, Karikari CA, et al. Array-based comparative genomic hybridization identifies localized DNA amplifications and homozygous deletions in pancreatic cancer. Neoplasia. 2005;7:556–62.
Cooke SL, Pole JC, Chin SF, Ellis IO, Caldas C, Edwards PA. High-resolution array CGH clarifies events occurring on 8p in carcinogenesis. BMC Cancer. 2008;8:288.
Harada T, Chelala C, Bhakta V, Chaplin T, Caulee K, Baril P, et al. Genome-wide DNA copy number analysis in pancreatic cancer using high-density single nucleotide polymorphism arrays. Oncogene. 2008;27:1951–60.
Harada T, Chelala C, Crnogorac-Jurcevic T, Lemoine NR. Genome-wide analysis of pancreatic cancer using microarray-based techniques. Pancreatology. 2009;9:13–24.
Williams SV, Platt FM, Hurst CD, Aveyard JS, Taylor CF, Pole JC, et al. High-resolution analysis of genomic alterationon chromosome arm 8p in urothelial carcinoma. Genes Chromosomes Cancer. 2010;49:642–59.
Xue W, Kitzing T, Roessler S, Zuber J, Krasnitz A, Schultz N, et al. A cluster of cooperating tumour-suppressor genes candidates in chromosomal deletions. Proc Natl Acad Sci USA. 2012;109:8212–7.
The Cancer Genome Atlas Research Network. Integrated genomic characterization of pancreatic ductal adenocarcinoma cancer. Cancer Cell. 2017;32:185–203.e13.
International Cancer Genome Consortium, Hudson TJ, Anderson W, Artez A, Barker AD, Bell C, et al. International network of cancer genome projects. Nature. 2010;464:993–8.
Subramanian A, Tamayo P, Mootha VK, Mukherjee S, Ebert BL, Gillette MA, et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci USA. 2005;102:15545–50.
Wang N, Liang H, Zhou Y, Wang C, Zhang S, Pan Y, et al. miRNA-203 suppresses proliferation and migration and promotes the apoptosis of lung cancer cells by targeting SRC. PLoS One. 2014;9:e105570.
Morton JP, Karim SA, Graham K, Timpson P, Jamieson N, Athineos D, et al. Dasatinib inhibits the development of metastases in a mouse model of pancreatic ductal adenocarcinoma. Gastroenterology. 2010;139:292–303.
Cai Y, Crowther J, Pastor T, Abbasi Asbagh L, Baietti MF, De Troyer M, et al. Loss of chromosome 8p governs tumor progression and drug response altering lipid metabolism. Cancer Cell. 2016;29:751–66.
Ridley AJ, Hall A. The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors. Cell. 1992;70:389–99.
Horiuchi A, Imai T, Wang C, Ohira S, Feng Y, Nikaido T, et al. Up-regulation of small GTPases, RhoA and RhoC, is associated with tumour progression in ovarian carcinoma. Lab Investig. 2003;83:861–70.
Barrio-Real L, Kazanietz MG. Rho GEFs and cancer: linking gene expression and metastatic dissemination. Sci Signal. 2012;5:pe43.
Konstantinidou G, Ramadori G, Torti F, Kangasniemi K, Ramirez RE, Cai Y, et al. RHOA-FAK is a required signaling axis for the maintenance of KRAS-driven lung adenocarcinomas. Cancer Discov. 2013;3:444–57.
Zandvakili I, Lin Y, Morris JC, Zheng Y. Rho GTPases: anti- or pro-neoplastic targets? Oncogene. 2017;36:3213–22.
Lazer G, Idelchuk Y, Schapira V, Pikarsky E, Katzav S. The haematopoietic specific signal transducer Vav1 is aberrantly expressed in lung cancer and plays a role in tumorigenesis. J Pathol. 2009;219:25–34.
Razidlo GL, Magnine C, Sletten AC, Hurley RM, Almada LL, Fernandez-Zapico ME, et al. Targeting pancreatic cancer metastasis by inhibition of Vav1, a driver of tumour cell invasion. Cancer Res. 2015;75:2907–15.
Jin H, Li T, Ding Y, Deng Y, Zhang W, Yang H, et al. Methylation status of T-lymphoma invasion and metastasis 1 promoter and its overexpression in colorectal cancer. Hum Pathol. 2011;42:541–51.
Qin J, Xie Y, Wang B, Hoshino M, Wolff DW, Zhao J, et al. Upregulation of P1P3-dependent Rac exchanger 1 (P-rex1) promotes prostate cancer metastasis. Oncogene. 2009;28:1853–63.
Liu Ax, Cerniglia GJ, Bernhard EJ, Prendergast GC. RhoB is required to mediate apoptosis in neoplastically transformed cells after DNA damage. Proc Natl Acad Sci USA. 2001;98:6192–7.
Adnane J, Muro-Cacho C, Mathews L, Sebti SM, Muñoz-Antonia T. Suppression of rho B expression in invasive carcinoma from head and neck cancer patients. Clin Cancer Res. 2002;8:2225–32.
Mazieres J, Antonia T, Daste G, Muro-Cacho C, Berchery D, Tillement V, et al. Loss of RhoB expression in human lung cancer progression. Clin Cancer Res. 2004;10:2742–50.
Marlow LA, Reynolds LA, Cleland AS, Cooper SJ, Gumz ML, Kurakata S, et al. Reactivation of suppressed RhoB is a critical step for the inhibition of anaplastic thyroid cancer growth. Cancer Res. 2009;69:1536–44.
Zhou J, Zhu Y, Zhang G, Liu N, Sun L, Liu M, et al. A distinct role of RhoB in gastric cancer suppression. Int J Cancer. 2011;128:1057–68.
Kazerounian S, Gerald D, Huang M, Chin YR, Udayakumar D, Zheng N, et al. RhoB differentially controls Akt function in tumour cells and stromal endothelial cells during breast cancer tumorigenesis. Cancer Res. 2013;73:50–61.
Tan Y, Yin H, Zhang H, Fang J, Zheng W, Li D, et al. Sp1-driven up-regulation of miR-119a decreases RHOB and promotes pancreatic cancer. Oncotarget. 2015;6:17391–403.
Robles-Valero J, Lorenzo-MartÃn LF, Menacho-Márquez M, Fernández-Pisonero I, Abad A, Camós M, et al. A paradoxical tumor-suppressor role for the Rac1 exchange factor Vav1 in T cell acute lymphoblastic leukemia. Cancer Cell. 2017;32:608–.e9.
Summy JM, Trevino JG, Baker CH, Gallick GE. C-Src regulates constitutive and EGF-mediated VEGF expression in pancreatic tumor cells through activation of phosphatidyl inositol-3 kinase and p38 MAPK. Pancreas. 2005;31:263–74.
Lopez J, Hesling C, Prudent J, Popgeorgiev N, Gadet R, Mikaelian I, et al. Src tyrosine kinase inhibits apoptosis through the Erk1/2-dependent degradation of the death accelerator Bik. Cell Death Differ. 2012;19:1459–69.
Lutz MP, Esser IB, Flossmann-Kast BB, Vogelmann R, Lührs H, Friess H, et al. Overexpression and activation of the tyrosine kinase Src in human pancreatic carcinoma. Biochem Biophys Res Commun. 1998;243:503–8.
Nagaraj NS, Smith JJ, Revetta F, Washington MK, Merchant NB. Targeted inhibition of SRC kinase signaling attenuates pancreatic tumorigenesis. Mol Cancer Ther. 2010;9:2322–32.
Shields DJ, Murphy EA, Desgrosellier JS, Mielgo A, Lau SK, Barnes LA, et al. Oncogenic Ras/Src cooperativity in pancreatic neoplasia. Oncogene. 2011;30:2123–34.
Witkiewicz AK, Balaji U, Eslinger C, McMillan E, Conway W, Posner B, et al. Integrated patient-derived models delineate individualized therapeutic vulnerabilities of pancreatic cancer. Cell Rep. 2016;16:2017–31.
Shi H, Zhang CJ, Chen GY, Yao SQ. Cell-based proteome profiling of potential dasatinib targets by use of affinity-based probes. J Am Chem Soc. 2012;134:3001–14.
Furukawa T, Duguid WP, Rosenberg L, Viallet J, Galloway DA, Tsao MS. Long-term culture and immortalization of epithelial cells from normal adult human pancreatic ducts transfected by the E6E7 gene of human papilloma virus 16. Am J Pathol. 1996;148:1763–70.
Qian J, Niu J, Li M, Chiao PJ, Tsao MS. In vitro modeling of human pancreatic duct epithelial cell transformation defines gene expression changes induced by K-ras oncogenic activation in pancreatic carcinogenesis. Cancer Res. 2005;12:5045–53.
Lohse I, Lourenco C, Ibrahimov E, Pintilie M, Tsao MS, Hedley DW. Assessment of hypoxia in the stroma of patient-derived pancreatic tumor xenografts. Cancers. 2014;6:459–71.
Mak AB, Ni Z, Hewel JA, Chen GI, Zhong G, Karamboulas K, et al. A lentiviral functional proteomics approach identifies chromatin remodeling complexes important for the induction of pluripotency. Mol Cell Proteom. 2010;9:811–23.
Abramoff MD, Magalhaes PJ, Ram SJ. Image processing with ImageJ. Biophotonics Int. 2004;11:36–42.
Acknowledgements
We thank Ming Li for his assistance with orthotopic implantation, Jing Xu for her assistance with immunohistochemistry, and Quan Li for his assistance with the analysis of gene expression data. This work was supported by the Canadian Cancer Society grant #700809, the Canadian Institutes of Health Research Foundation Scheme grant FDN-148395, and the Ontario Ministry of Health and Long Term Care. TW was supported by the Terry Fox Foundation Training Program in Molecular Pathology of Cancer at CIHR (STP 53912). Dr Tsao is the M. Qasim Choksi Chair in Lung Cancer Translational Research. The microarray data reported in this paper have been deposited into the Gene Expression Omnibus (GEO) database (accession number GSE131859).
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Joseph, J., Radulovich, N., Wang, T. et al. Rho guanine nucleotide exchange factor ARHGEF10 is a putative tumor suppressor in pancreatic ductal adenocarcinoma. Oncogene 39, 308–321 (2020). https://doi.org/10.1038/s41388-019-0985-1
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DOI: https://doi.org/10.1038/s41388-019-0985-1