Axitinib and crizotinib combination therapy inhibits bone loss in a mouse model of castration resistant prostate cancer

BMC Cancer

Jeetendra Eswaraka¹  Anand Giddabasappa¹  Guangzhou Han³  Kush Lalwani¹  Koleen Eisele³  Zheng Feng³  Timothy Affolter²  James Christensen³  Gang Li³ 

[1] Global Science and Technology (WCM), Pfizer Global Research and Development, 10724 Science Center Dr, San Diego, CA 92121, USA

[2] Drug Safety Research and Development Pfizer Inc, 10724 Science Center Drive, San Diego, CA 92121, USA

[3] Oncology Research Unit, Pfizer Global Research and Development, 10724 Science Center Dr, San Diego, CA 92121, USA

关键词: c-MET;    VEGFR crizotinib;    Axitinib;    BLI;    X-ray;    μCT imaging;    Bone metastasis;    CRPC;   

Others  :  1120998
DOI  :  10.1186/1471-2407-14-742

 received in 2014-07-11, accepted in 2014-09-23,  发布年份 2014

【 摘 要 】

Background

Castration resistant prostate cancer (CRPC) is a leading cause of cancer-related deaths in men. The primary cause of mortality and morbidity in patients is bone metastases and remodeling resulting in osteoblastic and osteolytic lesions. Recently, cabozantinib, a multi-kinase inhibitor (VEGFR2 and c-MET inhibitor), was shown to have efficacy on bone lesions in patients. In this study we tested multi-kinase inhibitors: axitinib (VEGFR inhibitor) and crizotinib (c-MET inhibitor) in a combination trial in mice models.

Methods

VCaP-Luc cells were grown as subcutaneous implants in intact and castrated NOD-SCID-gamma (NSG) mice to confirm the androgen dependency. For bone metastasis model two cohorts of NSG mice (castrated and intact) received orthotopic injection of VCaP-Luc cells into the bone marrow cavity of left tibia. Mice were monitored weekly for tumor growth using bioluminescence imaging. Animals were randomized into 4 groups based on the tumor bioluminescence signal: vehicle, crizotinib alone, axitinib alone, crizotinib and axitinib in combination. Animals were imaged weekly by in vivo 2-D X-ray imaging to monitor bone remodeling. At the end of the study animals were euthanized and both tibias were extracted for ex vivo high-resolution 3-D micro-computed tomography (μCT) imaging.

Results

Subcutaneous model showed that androgen stimulation may be helpful but not essential for the growth of VCaP-Luc cells. VCaP-Luc cells grown intra-tibially in intact animals caused extensive remodeling of bone with mixed osteoblastic (bone formation) and osteolytic (bone matrix dissolution) lesions. The osteoblastic lesions were predominant and at times extended beyond the tibial shaft into the surrounding tissue. In contrast, only osteolytic lesions were prominent throughout the study in castrated animals. Treatment with crizotinib alone reduced the osteolytic lesions in castrated animals. Axitinib alone reduced the osteoblastic lesions in the intact animals. Combination therapy with axitinib and crizotinib remarkably inhibited the tibial remodeling by VCaP-Luc cells which resulted in a significant reduction of both osteoblastic and osteolytic lesions.

Conclusion

Our data show that combined inhibition of c-MET and VEGFR can be beneficial for treatment of metastatic bone disease in CRPC and that the drugs act on two different stages of the disease.

【 授权许可】

   
2014 Eswaraka et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Jemal A, Siegel R, Xu J, Ward E: Cancer statistics, 2010. CA Cancer J Clin 2010, 60(5):277-300.
• [2]Walsh PC, DeWeese TL, Eisenberger MA: Clinical practice. localized prostate cancer. N Engl J Med 2007, 357(26):2696-2705.
• [3]Attard G, Sarker D, Reid A, Molife R, Parker C, de Bono JS: Improving the outcome of patients with castration-resistant prostate cancer through rational drug development. Br J Cancer 2006, 95(7):767-774.
• [4]Mehra R, Kumar-Sinha C, Shankar S, Lonigro RJ, Jing X, Philips NE, Siddiqui J, Han B, Cao X, Smith DC, Shah RB, Chinnaiyan AM, Pienta KJ: Characterization of bone metastases from rapid autopsies of prostate cancer patients. Clin Cancer Res 2011, 17(12):3924-3932.
• [5]Ohlmann CH, Merseburger AS, Suttmann H, Schilling D, Trojan L, Kempkensteffen C, Corvin S, Mathers MJ, Bastian PJ: Novel options for the treatment of castration-resistant prostate cancer. World J Urol 2012, 30(4):495-503.
• [6]Lee RJ, Saylor PJ, Smith MR: Treatment and prevention of bone complications from prostate cancer. Bone 2011, 48(1):88-95.
• [7]Sturge J, Caley MP, Waxman J: Bone metastasis in prostate cancer: emerging therapeutic strategies. Nat Rev Clin Oncol 2011, 8(6):357-368.
• [8]Bowles DW, Kessler ER, Jimeno A: Multi-targeted tyrosine kinase inhibitors in clinical development: focus on XL-184 (cabozantinib). Drugs Today (Barc) 2011, 47(11):857-868.
• [9]Yakes FM, Chen J, Tan J, Yamaguchi K, Shi Y, Yu P, Qian F, Chu F, Bentzien F, Cancilla B, Orf J, You A, Laird AD, Engst S, Lee L, Lesch J, Chou YC, Joly AH: Cabozantinib (XL184), a novel MET and VEGFR2 inhibitor, simultaneously suppresses metastasis, angiogenesis, and tumor growth. Mol Cancer Ther 2011, 10(12):2298-2308.
• [10]Duque JL, Loughlin KR, Adam RM, Kantoff PW, Zurakowski D, Freeman MR: Plasma levels of vascular endothelial growth factor are increased in patients with metastatic prostate cancer. Urology 1999, 54(3):523-527.
• [11]Weidner N, Carroll PR, Flax J, Blumenfeld W, Folkman J: Tumor angiogenesis correlates with metastasis in invasive prostate carcinoma. Am J Pathol 1993, 143(2):401-409.
• [12]Merino M, Pinto A, Gonzalez R, Espinosa E: Antiangiogenic agents and endothelin antagonists in advanced castration resistant prostate cancer. Eur J Cancer 2011, 47(12):1846-1851.
• [13]Mayr-Wohlfart U, Waltenberger J, Hausser H, Kessler S, Gunther KP, Dehio C, Puhl W, Brenner RE: Vascular endothelial growth factor stimulates chemotactic migration of primary human osteoblasts. Bone 2002, 30(3):472-477.
• [14]Midy V, Plouet J: Vasculotropin/vascular endothelial growth factor induces differentiation in cultured osteoblasts. Biochem Biophys Res Commun 1994, 199(1):380-386.
• [15]Dai J, Kitagawa Y, Zhang J, Yao Z, Mizokami A, Cheng S, Nor J, McCauley LK, Taichman RS, Keller ET: Vascular endothelial growth factor contributes to the prostate cancer-induced osteoblast differentiation mediated by bone morphogenetic protein. Cancer Res 2004, 64(3):994-999.
• [16]Kitagawa Y, Dai J, Zhang J, Keller JM, Nor J, Yao Z, Keller ET: Vascular endothelial growth factor contributes to prostate cancer-mediated osteoblastic activity. Cancer Res 2005, 65(23):10921-10929.
• [17]van Leenders G, van Balken B, Aalders T, Hulsbergen-van de Kaa C, Ruiter D, Schalken J: Intermediate cells in normal and malignant prostate epithelium express c-MET: implications for prostate cancer invasion. Prostate 2002, 51(2):98-107.
• [18]Humphrey PA, Zhu X, Zarnegar R, Swanson PE, Ratliff TL, Vollmer RT, Day ML: Hepatocyte growth factor and its receptor (c-MET) in prostatic carcinoma. Am J Pathol 1995, 147(2):386-396.
• [19]Knudsen BS, Gmyrek GA, Inra J, Scherr DS, Vaughan ED, Nanus DM, Kattan MW, Gerald WL, Vande Woude GF: High expression of the Met receptor in prostate cancer metastasis to bone. Urology 2002, 60(6):1113-1117.
• [20]Colombel M, Eaton CL, Hamdy F, Ricci E, van der Pluijm G, Cecchini M, Mege-Lechevallier F, Clezardin P, Thalmann G: Increased expression of putative cancer stem cell markers in primary prostate cancer is associated with progression of bone metastases. Prostate 2012, 72(7):713-720.
• [21]Russo AL, Jedlicka K, Wernick M, McNally D, Kirk M, Sproull M, Smith S, Shankavaram U, Kaushal A, Figg WD, Dahut W, Citrin D, Bottaro DP, Albert PS, Tofilon PJ, Camphausen K: Urine analysis and protein networking identify met as a marker of metastatic prostate cancer. Clin Cancer Res 2009, 15(13):4292-4298.
• [22]Verras M, Lee J, Xue H, Li TH, Wang Y, Sun Z: The androgen receptor negatively regulates the expression of c-Met: implications for a novel mechanism of prostate cancer progression. Cancer Res 2007, 67(3):967-975.
• [23]Hu-Lowe DD, Zou HY, Grazzini ML, Hallin ME, Wickman GR, Amundson K, Chen JH, Rewolinski DA, Yamazaki S, Wu EY, McTigue MA, Murray BW, Kania RS, O'Connor P, Shalinsky DR, Bender SL: Nonclinical antiangiogenesis and antitumor activities of axitinib (AG-013736), an oral, potent, and selective inhibitor of vascular endothelial growth factor receptor tyrosine kinases 1, 2, 3. Clin Cancer Res 2008, 14(22):7272-7283.
• [24]Cui JJ, Tran-Dube M, Shen H, Nambu M, Kung PP, Pairish M, Jia L, Meng J, Funk L, Botrous I, McTigue M, Grodsky N, Ryan K, Padrique E, Alton G, Timofeevski S, Yamazaki S, Li Q, Zou H, Christensen J, Mroczkowski B, Bender S, Kania RS, Edwards MP: Structure based drug design of crizotinib (PF-02341066), a potent and selective dual inhibitor of mesenchymal-epithelial transition factor (c-MET) kinase and anaplastic lymphoma kinase (ALK). J Med Chem 2011, 54(18):6342-6363.
• [25]Li X, Loberg R, Liao J, Ying C, Snyder LA, Pienta KJ, McCauley LK: A destructive cascade mediated by CCL2 facilitates prostate cancer growth in bone. Cancer Res 2009, 69(4):1685-1692.
• [26]Park SI, Kim SJ, McCauley LK, Gallick GE: Pre-clinical mouse models of human prostate cancer and their utility in drug discovery. Curr Protoc Pharmacol 2010, Chapter 14:Unit 14 15.
• [27]Corey E, Quinn JE, Bladou F, Brown LG, Roudier MP, Brown JM, Buhler KR, Vessella RL: Establishment and characterization of osseous prostate cancer models: intra-tibial injection of human prostate cancer cells. Prostate 2002, 52(1):20-33.
• [28]Siegel R, Naishadham D, Jemal A: Cancer statistics, 2013. CA Cancer J Clin 2013, 63(1):11-30.
• [29]Chi KN, Bjartell A, Dearnaley D, Saad F, Schroder FH, Sternberg C, Tombal B, Visakorpi T: Castration-resistant prostate cancer: from new pathophysiology to new treatment targets. Eur Urol 2009, 56(4):594-605.
• [30]Scher HI, Halabi S, Tannock I, Morris M, Sternberg CN, Carducci MA, Eisenberger MA, Higano C, Bubley GJ, Dreicer R, Petrylak D, Kantoff P, Basch E, Kelly WK, Figg WD, Small EJ, Beer TM, Wilding G, Martin A, Hussain M, Prostate Cancer Clinical Trials Working G: Design and end points of clinical trials for patients with progressive prostate cancer and castrate levels of testosterone: recommendations of the Prostate Cancer Clinical Trials Working Group. J Clin Oncol 2008, 26(7):1148-1159.
• [31]Attar RM, Takimoto CH, Gottardis MM: Castration-resistant prostate cancer: locking up the molecular escape routes. Clin Cancer Res 2009, 15(10):3251-3255.
• [32]Scher HI, Sawyers CL: Biology of progressive, castration-resistant prostate cancer: directed therapies targeting the androgen-receptor signaling axis. J Clin Oncol 2005, 23(32):8253-8261.
• [33]Coleman RE: Metastatic bone disease: clinical features, pathophysiology and treatment strategies. Cancer Treat Rev 2001, 27(3):165-176.
• [34]Ibrahim T, Flamini E, Mercatali L, Sacanna E, Serra P, Amadori D: Pathogenesis of osteoblastic bone metastases from prostate cancer. Cancer 2010, 116(6):1406-1418.
• [35]Bagi CM: Skeletal implications of prostate cancer. J Musculoskelet Neuronal Interact 2003, 3(2):112-117.
• [36]Keller ET, Brown J: Prostate cancer bone metastases promote both osteolytic and osteoblastic activity. J Cell Biochem 2004, 91(4):718-729.
• [37]Logothetis CJ, Lin SH: Osteoblasts in prostate cancer metastasis to bone. Nat Rev Cancer 2005, 5(1):21-28.
• [38]DePuy V, Anstrom KJ, Castel LD, Schulman KA, Weinfurt KP, Saad F: Effects of skeletal morbidities on longitudinal patient-reported outcomes and survival in patients with metastatic prostate cancer. Support Care Cancer 2007, 15(7):869-876.
• [39]Lage MJ, Barber BL, Harrison DJ, Jun S: The cost of treating skeletal-related events in patients with prostate cancer. Am J Manag Care 2008, 14(5):317-322.
• [40]Saad F, Gleason DM, Murray R, Tchekmedyian S, Venner P, Lacombe L, Chin JL, Vinholes JJ, Goas JA, Zheng M, Zoledronic Acid Prostate Cancer Study G: Long-term efficacy of zoledronic acid for the prevention of skeletal complications in patients with metastatic hormone-refractory prostate cancer. J Natl Cancer Inst 2004, 96(11):879-882.
• [41]Fizazi K, Carducci M, Smith M, Damiao R, Brown J, Karsh L, Milecki P, Shore N, Rader M, Wang H, Jiang Q, Tadros S, Dansey R, Goessl C: Denosumab versus zoledronic acid for treatment of bone metastases in men with castration-resistant prostate cancer: a randomised, double-blind study. Lancet 2011, 377(9768):813-822.
• [42]Smith DC, Smith MR, Sweeney C, Elfiky AA, Logothetis C, Corn PG, Vogelzang NJ, Small EJ, Harzstark AL, Gordon MS, Vaishampayan UN, Haas NB, Spira AI, Lara PN Jr, Lin CC, Srinivas S, Sella A, Schoffski P, Scheffold C, Weitzman AL, Hussain M: Cabozantinib in patients with advanced prostate cancer: results of a phase II randomized discontinuation trial. J Clin Oncol 2013, 31(4):412-419.
• [43]Birchmeier C, Birchmeier W, Gherardi E, Vande Woude GF: Met, metastasis, motility and more. Nat Rev Mol Cell Biol 2003, 4(12):915-925.
• [44]Zou HY, Li Q, Lee JH, Arango ME, McDonnell SR, Yamazaki S, Koudriakova TB, Alton G, Cui JJ, Kung PP, Nambu MD, Los G, Bender SL, Mroczkowski B, Christensen JG: An orally available small-molecule inhibitor of c-Met, PF-2341066, exhibits cytoreductive antitumor efficacy through antiproliferative and antiangiogenic mechanisms. Cancer Res 2007, 67(9):4408-4417.
• [45]Pfeiffer MJ, Smit FP, Sedelaar JP, Schalken JA: Steroidogenic enzymes and stem cell markers are upregulated during androgen deprivation in prostate cancer. Mol Med 2011, 17(7–8):657-664.
• [46]Grano M, Galimi F, Zambonin G, Colucci S, Cottone E, Zallone AZ, Comoglio PM: Hepatocyte growth factor is a coupling factor for osteoclasts and osteoblasts in vitro. Proc Natl Acad Sci U S A 1996, 93(15):7644-7648.
• [47]Sato T, Hakeda Y, Yamaguchi Y, Mano H, Tezuka K, Matsumoto K, Nakamura T, Mori Y, Yoshizawa K, Sumitani K: Hepatocyte growth factor is involved in formation of osteoclast-like cells mediated by clonal stromal cells (MC3T3-G2/PA6). J Cell Physiol 1995, 164(1):197-204.
• [48]George DJ, Halabi S, Shepard TF, Vogelzang NJ, Hayes DF, Small EJ, Kantoff PW, Cancer, Leukemia Group B: Prognostic significance of plasma vascular endothelial growth factor levels in patients with hormone-refractory prostate cancer treated on Cancer and Leukemia Group B 9480. Clin Cancer Res 2001, 7(7):1932-1936.
• [49]Street J, Lenehan B: Vascular endothelial growth factor regulates osteoblast survival - evidence for an autocrine feedback mechanism. J Orthop Surg Res 2009, 4:19. BioMed Central Full Text