Abstract
Antiangiogenic therapy resistance occurs frequently in patients with metastatic renal cell carcinoma (RCC). The purpose of this study was to understand the mechanism of resistance to sunitinib, an antiangiogenic small molecule, and to exploit this mechanism therapeutically. We hypothesized that sunitinib-induced upregulation of the prometastatic MET and AXL receptors is associated with resistance to sunitinib and with more aggressive tumor behavior. In the present study, tissue microarrays containing sunitinib-treated and untreated RCC tissues were stained with MET and AXL antibodies. The low malignant RCC cell line 786-O was chronically treated with sunitinib and assayed for AXL, MET, epithelial–mesenchymal transition (EMT) protein expression and activation. Co-culture experiments were used to examine the effect of sunitinib pretreatment on endothelial cell growth. The effects of AXL and MET were evaluated in various cell-based models by short hairpin RNA or inhibition by cabozantinib, the multi-tyrosine kinases inhibitor that targets vascular endothelial growth factor receptor, MET and AXL. Xenograft mouse models tested the ability of cabozantinib to rescue sunitinib resistance. We demonstrated that increased AXL and MET expression was associated with inferior clinical outcome in patients. Chronic sunitinib treatment of RCC cell lines activated both AXL and MET, induced EMT-associated gene expression changes, including upregulation of Snail and β-catenin, and increased cell migration and invasion. Pretreatment with sunitinib enhanced angiogenesis in 786-0/human umbilical vein endothelial cell co-culture models. The suppression of AXL or MET expression and the inhibition of AXL and MET activation using cabozantinib both impaired chronic sunitinib treatment-induced prometastatic behavior in cell culture and rescued acquired resistance to sunitinib in xenograft models. In summary, chronic sunitinib treatment induces the activation of AXL and MET signaling and promotes prometastatic behavior and angiogenesis. The inhibition of AXL and MET activity may overcome resistance induced by prolonged sunitinib therapy in metastatic RCC.
References
- 1.
Thoenes W, Storkel S, Rumpelt HJ. Histopathology and classification of renal cell tumors (adenomas, oncocytomas and carcinomas). The basic cytological and histopathological elements and their use for diagnostics. Pathol Res Pract 1986; 181: 125–143.
- 2.
Kanesvaran R, Tan MH. Targeted therapy for renal cell carcinoma: The next lap. J Carcinog 2014; 13: 3.
- 3.
Domblides C, Gross-Goupil M, Quivy A, Ravaud A. Emerging antiangiogenics for renal cancer. Expert Opin Emerg Drugs 2013; 18: 495–511.
- 4.
Griffioen AW, Mans LA, de Graaf AM, Nowak-Sliwinska P, de Hoog CL, de Jong TA et al. Rapid angiogenesis onset after discontinuation of sunitinib treatment of renal cell carcinoma patients. Clin Cancer Res 2012; 18: 3961–3971.
- 5.
Huang D, Ding Y, Zhou M, Rini BI, Petillo D, Qian CN et al. Interleukin-8 mediates resistance to antiangiogenic agent sunitinib in renal cell carcinoma. Cancer Res 2010; 70: 1063–1071.
- 6.
Jonasch E, Corn P, Pagliaro LC, Warneke CL, Johnson MM, Tamboli P et al. Upfront, randomized, phase 2 trial of sorafenib versus sorafenib and low-dose interferon alfa in patients with advanced renal cell carcinoma: clinical and biomarker analysis. Cancer 2010; 116: 57–65.
- 7.
Tsavachidou-Fenner D, Tannir N, Tamboli P, Liu W, Petillo D, Teh B et al. Gene and protein expression markers of response to combined antiangiogenic and epidermal growth factor targeted therapy in renal cell carcinoma. Ann Oncol 2010; 21: 1599–1606.
- 8.
Kim J, Jonasch E, Alexander A, Short JD, Cai S, Wen S et al. Cytoplasmic sequestration of p27 via AKT phosphorylation in renal cell carcinoma. Clin Cancer Res 2009; 15: 81–90.
- 9.
Rankin EB, Fuh KC, Castellini L, Viswanathan K, Finger EC, Diep AN et al. Direct regulation of GAS6/AXL signaling by HIF promotes renal metastasis through SRC and MET. Proc Natl Acad Sci USA 2014; 111: 13373–13378.
- 10.
Peters S, Adjei AA. MET: a promising anticancer therapeutic target. Nat Rev Clin Oncol 2012; 9: 314–326.
- 11.
Birchmeier C, Birchmeier W, Gherardi E, Vande Woude GF. Met metastasis, motility and more. Nat Rev Mol Cell Biol 2003; 4: 915–925.
- 12.
Gherardi E, Birchmeier W, Birchmeier C, Vande Woude G. Targeting MET in cancer: rationale and progress. Nat Rev Cancer 2012; 12: 89–103.
- 13.
Gibney GT, Aziz SA, Camp RL, Conrad P, Schwartz BE, Chen CR et al. c-Met is a prognostic marker and potential therapeutic target in clear cell renal cell carcinoma. Ann Oncol 2013; 24: 343–349.
- 14.
Miyata Y, Kanetake H, Kanda S. Presence of phosphorylated hepatocyte growth factor receptor/c-Met is associated with tumor progression and survival in patients with conventional renal cell carcinoma. Clin Cancer Res 2006; 12: 4876–4881.
- 15.
Betsunoh H, Mukai S, Akiyama Y, Fukushima T, Minamiguchi N, Hasui Y et al. Clinical relevance of hepsin and hepatocyte growth factor activator inhibitor type 2 expression in renal cell carcinoma. Cancer Sci 2007; 98: 491–498.
- 16.
Sennino B, Ishiguro-Oonuma T, Wei Y, Naylor RM, Williamson CW, Bhagwandin V et al. Suppression of tumor invasion and metastasis by concurrent inhibition of c-Met and VEGF signaling in pancreatic neuroendocrine tumors. Cancer Discov 2012; 2: 270–287.
- 17.
Rankin EB, Fuh KC, Taylor TE, Krieg AJ, Musser M, Yuan J et al. AXL is an essential factor and therapeutic target for metastatic ovarian cancer. Cancer Res 2010; 70: 7570–7579.
- 18.
Ye X, Li Y, Stawicki S, Couto S, Eastham-Anderson J, Kallop D et al. An anti-Axl monoclonal antibody attenuates xenograft tumor growth and enhances the effect of multiple anticancer therapies. Oncogene 2010; 29: 5254–5264.
- 19.
Holland SJ, Powell MJ, Franci C, Chan EW, Friera AM, Atchison RE et al. Multiple roles for the receptor tyrosine kinase axl in tumor formation. Cancer Res 2005; 65: 9294–9303.
- 20.
Linger RM, Keating AK, Earp HS, Graham DK. TAM receptor tyrosine kinases: biologic functions, signaling, and potential therapeutic targeting in human cancer. Adv Cancer Res 2008; 100: 35–83.
- 21.
Linger RM, Keating AK, Earp HS, Graham DK. Taking aim at Mer and Axl receptor tyrosine kinases as novel therapeutic targets in solid tumors. Expert Opin Ther Targets 2010; 14: 1073–1090.
- 22.
Gujral TS, Karp RL, Finski A, Chan M, Schwartz PE, Macbeath G et al. Profiling phospho-signaling networks in breast cancer using reverse-phase protein arrays. Oncogene 2012; 32: 3470–3476.
- 23.
Zhang Z, Lee JC, Lin L, Olivas V, Au V, Laframboise T et al. Activation of the AXL kinase causes resistance to EGFR-targeted therapy in lung cancer. Nat Genet 2012; 44: 852–860.
- 24.
Ruan GX, Kazlauskas A. Axl is essential for VEGF-A-dependent activation of PI3K/Akt. EMBO J 2012; 31: 1692–1703.
- 25.
Chung BI, Malkowicz SB, Nguyen TB, Libertino JA, McGarvey TW. Expression of the proto-oncogene Axl in renal cell carcinoma. DNA Cell Biol 2003; 22: 533–540.
- 26.
Gustafsson A, Martuszewska D, Johansson M, Ekman C, Hafizi S, Ljungberg B et al. Differential expression of Axl and Gas6 in renal cell carcinoma reflecting tumor advancement and survival. Clin Cancer Res 2009; 15: 4742–4749.
- 27.
Pao-Chun L, Chan PM, Chan W, Manser E. Cytoplasmic ACK1 interaction with multiple receptor tyrosine kinases is mediated by Grb2: an analysis of ACK1 effects on Axl signaling. J Biol Chem 2009; 284: 34954–34963.
- 28.
Leconet W, Larbouret C, Chardes T, Thomas G, Neiveyans M, Busson M et al. Preclinical validation of AXL receptor as a target for antibody-based pancreatic cancer immunotherapy. Oncogene 2014; 33: 5405–5414.
- 29.
Gustafsson A, Bostrom AK, Ljungberg B, Axelson H, Dahlback B. Gas6 and the receptor tyrosine kinase Axl in clear cell renal cell carcinoma. PLoS One 2009; 4: e7575.
- 30.
Horie S, Aruga S, Kawamata H, Okui N, Kakizoe T, Kitamura T. Biological role of HGF/MET pathway in renal cell carcinoma. J Urol 1999; 161: 990–997.
- 31.
Ellison AR, Lofing J, Bitter GA. Potentiation of human estrogen receptor alpha-mediated gene expression by steroid receptor coactivator-1 (SRC-1) in Saccharomyces cerevisiae. J Steroid Biochem Mol Biol 2003; 86: 15–26.
- 32.
Rahimi N, Hung W, Tremblay E, Saulnier R, Elliott B. c-Src kinase activity is required for hepatocyte growth factor-induced motility and anchorage-independent growth of mammary carcinoma cells. J Biol Chem 1998; 273: 33714–33721.
- 33.
Singhal E, Sen P. Hepatocyte growth factor-induced c-Src-phosphatidylinositol 3-kinase-AKT-mammalian target of rapamycin pathway inhibits dendritic cell activation by blocking IkappaB kinase activity. Int J Biochem Cell Biol 2011; 43: 1134–1146.
- 34.
Ho MY, Tang SJ, Chuang MJ, Cha TL, Li JY, Sun GH et al. TNF-alpha induces epithelial-mesenchymal transition of renal cell carcinoma cells via a GSK3beta-dependent mechanism. Mol Cancer Res 2012; 10: 1109–1119.
- 35.
Garofalo M, Romano G, Di Leva G, Nuovo G, Jeon YJ, Ngankeu A et al. EGFR and MET receptor tyrosine kinase-altered microRNA expression induces tumorigenesis and gefitinib resistance in lung cancers. Nat Med 2012; 18: 74–82.
- 36.
Friedl P, Alexander S. Cancer invasion and the microenvironment: plasticity and reciprocity. Cell 2011; 147: 992–1009.
- 37.
Harshman LC, Choueiri TK. Targeting the hepatocyte growth factor/c-Met signaling pathway in renal cell carcinoma. Cancer J 2013; 19: 316–323.
- 38.
Chinchar E, Makey KL, Gibson J, Chen F, Cole SA, Megason GC et al. Sunitinib significantly suppresses the proliferation, migration, apoptosis resistance, tumor angiogenesis and growth of triple-negative breast cancers but increases breast cancer stem cells. Vasc Cell 2014; 6: 12.
- 39.
Choueiri TK, Pal SK, McDermott DF, Morrissey S, Ferguson KC, Holland J et al. A phase I study of cabozantinib (XL184) in patients with renal cell cancer. Ann Oncol 2014; 25: 1603–1608.
- 40.
Bentzien F, Zuzow M, Heald N, Gibson A, Shi Y, Goon L et al. In vitro and in vivo activity of cabozantinib (XL184), an inhibitor of RET, MET, and VEGFR2, in a model of medullary thyroid cancer. Thyroid 2013; 23: 1569–1577.
- 41.
Zhang Y, Guessous F, Kofman A, Schiff D, Abounader R. XL-184, a MET, VEGFR-2 and RET kinase inhibitor for the treatment of thyroid cancer, glioblastoma multiforme and NSCLC. IDrugs 2010; 13: 112–121.
- 42.
Paez-Ribes M, Allen E, Hudock J, Takeda T, Okuyama H, Vinals F et al. Antiangiogenic therapy elicits malignant progression of tumors to increased local invasion and distant metastasis. Cancer Cell 2009; 15: 220–231.
- 43.
Kumar R, Crouthamel MC, Rominger DH, Gontarek RR, Tummino PJ, Levin RA et al. Myelosuppression and kinase selectivity of multikinase angiogenesis inhibitors. Br J Cancer 2009; 101: 1717–1723.
- 44.
Zhang Z, Lee JC, Lin L, Olivas V, Au V, LaFramboise T et al. Activation of the AXL kinase causes resistance to EGFR-targeted therapy in lung cancer. Nat Genet 2012; 44: 852–860.
- 45.
Plimack ER, Tannir N, Lin E, Bekele BN, Jonasch E. Patterns of disease progression in metastatic renal cell carcinoma patients treated with antivascular agents and interferon: impact of therapy on recurrence patterns and outcome measures. Cancer 2009; 115: 1859–1866.
- 46.
Smith DC, Smith MR, Sweeney C, Elfiky AA, Logothetis C, Corn PG et al. Cabozantinib in patients with advanced prostate cancer: results of a phase II randomized discontinuation trial. J Clin Oncol 2013; 31: 412–419.
- 47.
Jonasch E, Wood CG, Matin SF, Tu SM, Pagliaro LC, Corn PG et al. Phase II presurgical feasibility study of bevacizumab in untreated patients with metastatic renal cell carcinoma. J Clin Oncol 2009; 27: 4076–4081.
- 48.
Weidner N. Intratumor microvessel density as a prognostic factor in cancer. Am J Pathol 1995; 147: 9–19.
Acknowledgements
This work was supported by Exelixis, Renee Kaye Cure Fur Cancer Foundation, the Monteleone Family Foundation and the UT MD Anderson Cancer Center P30 Cancer Center Support Grant.
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Affiliations
Department of Genitourinary Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA
- L Zhou
- , X-D Liu
- , M Sun
- , X Zhang
- , P German
- , S Bai
- , Z Ding
- , N Tannir
- & E Jonasch
Department of Urology, Division of Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA
- C G Wood
- , S F Matin
- & J A Karam
Department of Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA
- P Tamboli
- , K Sircar
- , P Rao
- & A G Hoang
Department of Radiation Oncology, Stanford University School of Medicine, Stanford, CA, USA
- E B Rankin
- & A J Giaccia
Exelixis Inc., South San Francisco, CA, USA
- D A Laird
Institute of Biosciences and Technology, Texas A&M Health Science Center, Houston, TX, USA
- C L Walker
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Competing interests
Dr Jonasch has received research funding from Pfizer, Novartis, GSK, Onyx and Exelixis. Dr Jonasch is a consultant for Pfizer, GSK, Novartis and Genentech. The other authors declare no conflict of interest.
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Correspondence to E Jonasch.
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https://doi.org/10.1038/onc.2015.343
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