Canine pulmonary adenocarcinoma tyrosine kinase receptor expression and phosphorylation

BMC Veterinary Research

Gwendolen Lorch²  Christopher Premanandan¹  Evan T Mariotti² 

[1] Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, OH 43210, USA;Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus, OH 43210, USA

关键词: Tyrosine kinase receptors;    Platelet-derived growth factor receptor;    Lung cancer;    Epidermal growth factor receptor;    Canine;   

Others  :  1119351
DOI  :  10.1186/1746-6148-10-19

 received in 2013-08-28, accepted in 2014-01-08,  发布年份 2014

【 摘 要 】

Background

This study evaluated tyrosine kinase receptor (TKR) expression and activation in canine pulmonary adenocarcinoma (cpAC) biospecimens. As histological similarities exist between human and cpAC, we hypothesized that cpACs will have increased TKR mRNA and protein expression as well as TKR phosphorylation. The molecular profile of cpAC has not been well characterized making the selection of therapeutic targets that would potentially have relevant biological activity impossible. Therefore, the objectives of this study were to define TKR expression and their phosphorylation state in cpAC as well as to evaluate the tumors for the presence of potential epidermal growth factor receptor (EGFR) tyrosine kinase activating mutations in exons 18–21. Immunohistochemistry (IHC) for TKR expression was performed using a tissue microarray (TMA) constructed from twelve canine tumors and companion normal lung samples. Staining intensities of the IHC were quantified by a veterinary pathologist as well as by two different digitalized algorithm image analyses software programs. An antibody array was used to evaluate TKR phosphorylation of the tumor relative to the TKR phosphorylation of normal tissues with the resulting spot intensities quantified using array analysis software. Each EGFR exon PCR product from all of the tumors and non-affected lung tissues were sequenced using sequencing chemistry and the sequencing reactions were run on automated sequencer. Sequence alignments were made to the National Center for Biotechnology Information canine EGFR reference sequence.

Results

The pro-angiogenic growth factor receptor, PDGFRα, had increased cpAC tumor mRNA, protein expression and phosphorylation when compared to the normal lung tissue biospecimens. Similar to human pulmonary adenocarcinoma, significant increases in cpAC tumor mRNA expression and receptor phosphorylation of the anaplastic lymphoma kinase (ALK) tyrosine receptor were present when compared to the corresponding normal lung tissue. The EGFR mRNA, protein expression and phosphorylation were not increased compared to the normal lung and no activating mutations were identified in exons 18–21.

Conclusions

Canine pulmonary adenocarcinoma TKRs are detected at both the mRNA and protein levels and are activated. Further investigation into the contribution of TKR activation in cpAC tumorigenesis is warranted.

【 授权许可】

   
2014 Mariotti et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

【 参考文献 】

• [1]Sabattini S, Mancini FR, Marconato L, Bacci B, Rossi F, Vignoli M, Bettini G: EGFR overexpression in canine primary lung cancer: pathogenetic implications and impact on survival. Vet Comp Oncol10.1111/vco.12002
• [2]Griffey SM, Kraegel SA, Madewell BR: Rapid detection of K-ras gene mutations in canine lung cancer using single-strand conformational polymorphism analysis. Carcinogenesis 1998, 19(6):959-963.
• [3]Tierney LA, Hahn FF, Lechner JF: p53, erbB-2 and K-ras gene alterations are rare in spontaneous and plutonium-239-induced canine lung neoplasia. Radiat Res 1996, 145(2):181-187.
• [4]Lemmon MA, Schlessinger J: Cell signaling by receptor tyrosine kinases. Cell 2010, 141(7):1117-1134.
• [5]da Cunha SG, Shepherd FA, Tsao MS: EGFR mutations and lung cancer. Annu Rev Pathol 2011, 6:49-69.
• [6]Hayat MJ, Howlader N, Reichman ME, Edwards BK: Cancer statistics, trends, and multiple primary cancer analyses from the Surveillance, Epidemiology, and End Results (SEER) Program. Oncologist 2007, 12(1):20-37.
• [7]Poirier VJ, Burgess KE, Adams WM, Vail DM: Toxicity, dosage, and efficacy of vinorelbine (Navelbine) in dogs with spontaneous neoplasia. J Vet Intern Med 2004, 18(4):536-539.
• [8]Mehlhaff CJ, Leifer CE, Patnaik AK, Schwarz PD: Surgical-treatment of primary pulmonary neoplasia in 15 dogs. J Am Anim Hosp Assoc 1984, 20(5):799-803.
• [9]Khanna C, Vail DM: Targeting the lung: preclinical and comparative evaluation of anticancer aerosols in dogs with naturally occurring cancers. Curr Cancer Drug Targets 2003, 3(4):265-273.
• [10]Hershey AE, Kurzman ID, Forrest LJ, Bohling CA, Stonerook M, Placke ME, Imondi AR, Vail DM: Inhalation chemotherapy for macroscopic primary or metastatic lung tumors: proof of principle using dogs with spontaneously occurring tumors as a model. Clin Cancer Res 1999, 5(9):2653-2659.
• [11]London CA, Hannah AL, Zadovoskaya R, Chien MB, Kollias-Baker C, Rosenberg M, Downing S, Post G, Boucher J, Shenoy N, et al.: Phase I dose-escalating study of SU11654, a small molecule receptor tyrosine kinase inhibitor, in dogs with spontaneous malignancies. Clin Cancer Res 2003, 9(7):2755-2768.
• [12]Gillett NA, Stegelmeier BL, Kelly G, Haley PJ, Hahn FF: Expression of epidermal growth factor receptor in plutonium-239-induced lung neoplasms in dogs. Vet Pathol 1992, 29(1):46-52.
• [13]Stout RD, Jiang C, Matta B, Tietzel I, Watkins SK, Suttles J: Macrophages sequentially change their functional phenotype in response to changes in microenvironmental influences. J Immunol 2005, 175(1):342-349.
• [14]Wyckoff J, Wang W, Lin EY, Wang Y, Pixley F, Stanley ER, Graf T, Pollard JW, Segall J, Condeelis J: A paracrine loop between tumor cells and macrophages is required for tumor cell migration in mammary tumors. Cancer Res 2004, 64(19):7022-7029.
• [15]Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ: Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 1997, 25(17):3389-3402.
• [16]Altschul SF, Wootton JC, Gertz EM, Agarwala R, Morgulis A, Schaffer AA, Yu YK: Protein database searches using compositionally adjusted substitution matrices. Febs J 2005, 272(20):5101-5109.
• [17]Raaberg L, Nexo E, Buckley S, Luo W, Snead ML, Warburton D: Epidermal growth factor transcription, translation, and signal transduction by rat type II pneumocytes in culture. Am J Respir Cell Mol Biol 1992, 6(1):44-49.
• [18]Miettinen PJ, Warburton D, Bu D, Zhao JS, Berger JE, Minoo P, Koivisto T, Allen L, Dobbs L, Werb Z, et al.: Impaired lung branching morphogenesis in the absence of functional EGF receptor. Dev Biol 1997, 186(2):224-236.
• [19]Cho J, Pastorino S, Zeng Q, Xu X, Johnson W, Vandenberg S, Verhaak R, Cherniack AD, Watanabe H, Dutt A, et al.: Glioblastoma-derived epidermal growth factor receptor carboxyl-terminal deletion mutants are transforming and are sensitive to EGFR-directed therapies. Cancer Res 2011, 71(24):7587-7596.
• [20]Yarden Y, Sliwkowski MX: Untangling the ErbB signalling network. Nat Rev Mol Cell Biol 2001, 2(2):127-137.
• [21]Lee-Hoeflich ST, Crocker L, Yao E, Pham T, Munroe X, Hoeflich KP, Sliwkowski MX, Stern HM: A central role for HER3 in HER2-amplified breast cancer: implications for targeted therapy. Cancer Res 2008, 68(14):5878-5887.
• [22]Holbro T, Beerli RR, Maurer F, Koziczak M, Barbas CF 3rd, Hynes NE: The ErbB2/ErbB3 heterodimer functions as an oncogenic unit: ErbB2 requires ErbB3 to drive breast tumor cell proliferation. Proc Natl Acad Sci USA 2003, 100(15):8933-8938.
• [23]Hsieh AC, Moasser MM: Targeting HER proteins in cancer therapy and the role of the non-target HER3. Br J Cancer 2007, 97(4):453-457.
• [24]Greulich H, Kaplan B, Mertins P, Chen TH, Tanaka KE, Yun CH, Zhang X, Lee SH, Cho J, Ambrogio L, et al.: Functional analysis of receptor tyrosine kinase mutations in lung cancer identifies oncogenic extracellular domain mutations of ERBB2. Proc Natl Acad Sci USA 2012, 109(36):14476-14481.
• [25]Chen Y, Takita J, Choi YL, Kato M, Ohira M, Sanada M, Wang L, Soda M, Kikuchi A, Igarashi T, et al.: Oncogenic mutations of ALK kinase in neuroblastoma. Nature 2008, 455(7215):971-974.
• [26]Cabezon-Gutierrez L, Khosravi-Shahi P, Diaz-Munoz-de-la-Espada VM, Carrion-Galindo JR, Erana-Tomas I, Castro-Otero M: ALK-mutated non-small-cell lung cancer: a new strategy for cancer treatment. Lung 2012, 190(4):381-388.
• [27]Nitta H, Tsuta K, Yoshida A, Ho SN, Kelly BD, Murata LB, Kosmeder J, White K, Ehser S, Towne P, et al.: New methods for ALK status diagnosis in Non-small-cell lung cancer: an improved ALK immunohistochemical assay and a New, brightfield, dual ALK IHC-in situ hybridization assay. J Thorac Oncol 2013, 8(8):1019-1031.
• [28]Thunnissen E, Bubendorf L, Dietel M, Elmberger G, Kerr K, Lopez-Rios F, Moch H, Olszewski W, Pauwels P, Penault-Llorca F, et al.: EML4-ALK testing in non-small cell carcinomas of the lung: a review with recommendations. Virchows Arch 2012, 461(3):245-257.
• [29]Alvarez RH, Kantarjian HM, Cortes JE: Biology of platelet-derived growth factor and its involvement in disease. Mayo Clin Proc 2006, 81(9):1241-1257.
• [30]Dong J, Grunstein J, Tejada M, Peale F, Frantz G, Liang WC, Bai W, Yu L, Kowalski J, Liang X, et al.: VEGF-null cells require PDGFR alpha signaling-mediated stromal fibroblast recruitment for tumorigenesis. Embo J 2004, 23(14):2800-2810.
• [31]Chatterjee S, Heukamp LC, Siobal M, Schottle J, Wieczorek C, Peifer M, Frasca D, Koker M, Konig K, Meder L, et al.: Tumor VEGF:VEGFR2 autocrine feed-forward loop triggers angiogenesis in lung cancer. J Clin Invest 2013, 123(4):1732-1740.
• [32]Grossmann C, Freudinger R, Mildenberger S, Krug AW, Gekle M: Evidence for epidermal growth factor receptor as negative-feedback control in aldosterone-induced Na + reabsorption. Am J Physiol Renal Physiol 2004, 286(6):F1226-1231.
• [33]Wang Y, Pennock S, Chen X, Wang Z: Endosomal signaling of epidermal growth factor receptor stimulates signal transduction pathways leading to cell survival. Mol Cell Biol 2002, 22(20):7279-7290.
• [34]Newman SJ, Mrkonjich L, Walker KK, Rohrbach BW: Canine subcutaneous mast cell tumour: diagnosis and prognosis. J Comp Pathol 2007, 136(4):231-239.
• [35]Higgins RJ, Dickinson PJ, LeCouteur RA, Bollen AW, Wang H, Corely LJ, Moore LM, Zang W, Fuller GN: Spontaneous canine gliomas: overexpression of EGFR, PDGFRalpha and IGFBP2 demonstrated by tissue microarray immunophenotyping. J Neurooncol 2010, 98(1):49-55.
• [36]Brown RJ, Newman SJ, Durtschi DC, Leblanc AK: Expression of PDGFR-beta and Kit in canine anal sac apocrine gland adenocarcinoma using tissue immunohistochemistry. Vet Comp Oncol 2012, 10(1):74-79.
• [37]Yonemaru K, Sakai H, Murakami M, Yanai T, Masegi T: Expression of vascular endothelial growth factor, basic fibroblast growth factor, and their receptors (flt-1, flk-1, and flg-1) in canine vascular tumors. Vet Pathol 2006, 43(6):971-980.
• [38]Restucci B, Borzacchiello G, Maiolino P, Martano M, Paciello O, Papparella S: Expression of vascular endothelial growth factor receptor Flk-1 in canine mammary tumours. J Comp Pathol 2004, 130(2–3):99-104.
• [39]Giantin M, Aresu L, Benali S, Arico A, Morello EM, Martano M, Vascellari M, Castagnaro M, Lopparelli RM, Zancanella V, et al.: Expression of matrix metalloproteinases, tissue inhibitors of metalloproteinases and vascular endothelial growth factor in canine mast cell tumours. J Comp Pathol 2012, 147(4):419-429.
• [40]Urie BK, Russell DS, Kisseberth WC, London CA: Evaluation of expression and function of vascular endothelial growth factor receptor 2, platelet derived growth factor receptors-alpha and -beta, KIT, and RET in canine apocrine gland anal sac adenocarcinoma and thyroid carcinoma. BMC Vet Res 2012, 8:67. BioMed Central Full Text
• [41]Simpson RM, Bastian BC, Michael HT, Webster JD, Prasad ML, Conway CM, Prieto VM, Gary JM, Goldschmidt MH, Esplin DG, et al.: Sporadic naturally occurring melanoma in dogs as a preclinical model for human melanoma. Pigment Cell Melanoma Res 2014, 27(1):37-47.
• [42]Chevalier S, Defoy I, Lacoste J, Hamel L, Guy L, Begin LR, Aprikian AG: Vascular endothelial growth factor and signaling in the prostate: more than angiogenesis. Mol Cell Endocrinol 2002, 189(1–2):169-179.
• [43]Walker-Daniels J, Coffman K, Azimi M, Rhim JS, Bostwick DG, Snyder P, Kerns BJ, Waters DJ, Kinch MS: Overexpression of the EphA2 tyrosine kinase in prostate cancer. Prostate 1999, 41(4):275-280.