Tumor- and osteoclast-derived NRP2 in prostate cancer bone metastases

Abstract

Understanding the role of neuropilin 2 (NRP2) in prostate cancer cells as well as in the bone microenvironment is pivotal in the development of an effective targeted therapy for the treatment of prostate cancer bone metastasis. We observed a significant upregulation of NRP2 in prostate cancer cells metastasized to bone. Here, we report that targeting NRP2 in cancer cells can enhance taxane-based chemotherapy with a better therapeutic outcome in bone metastasis, implicating NRP2 as a promising therapeutic target. Since, osteoclasts present in the tumor microenvironment express NRP2, we have investigated the potential effect of targeting NRP2 in osteoclasts. Our results revealed NRP2 negatively regulates osteoclast differentiation and function in the presence of prostate cancer cells that promotes mixed bone lesions. Our study further delineated the molecular mechanisms by which NRP2 regulates osteoclast function. Interestingly, depletion of NRP2 in osteoclasts in vivo showed a decrease in the overall prostate tumor burden in the bone. These results therefore indicate that targeting NRP2 in prostate cancer cells as well as in the osteoclastic compartment can be beneficial in the treatment of prostate cancer bone metastasis.

References

[1] Macedo F et al. Bone metastases: an overview. Oncol. Rev., 2017, 11: 321

[2] Halabi S et al. Meta-analysis evaluating the impact of site of metastasis on overall survival in men with castration-resistant prostate cancer. J. Clin. Oncol., 2016, 34: 1652-1659

[3] Lund L, Borre M, Jacobsen J, Sorensen HT, Norgaard M. Impact of comorbidity on survival of Danish prostate cancer patients, 1995-2006: a population-based cohort study. Urology, 2008, 72: 1258-1262

[4] Terris, M. K., Qureshi, S. M. Metastatic and advanced prostate cancer. https://emedicine.medscape.com/article/454114-overview.

[5] Croucher PI, McDonald MM, Martin TJ. Bone metastasis: the importance of the neighbourhood. Nat. Rev. Cancer, 2016, 16: 373-386

[6] El-Amm J, Aragon-Ching JB. Targeting bone metastases in metastatic castration-resistant prostate cancer. Clin. Med. Insights Oncol., 2016, 10: 11-19

[7] Bienz M, Saad F. Management of bone metastases in prostate cancer: a review. Curr. Opin. Support Palliat. Care, 2015, 9: 261-267

[8] Merseburger AS et al. Perspectives on treatment of metastatic castration-resistant prostate cancer. Oncologist, 2013, 18: 558-567

[9] Sturge J, Caley MP, Waxman J. Bone metastasis in prostate cancer: emerging therapeutic strategies. Nat. Rev. Clin. Oncol., 2011, 8: 357-368

[10] Logothetis CJ, Lin SH. Osteoblasts in prostate cancer metastasis to bone. Nat. Rev. Cancer, 2005, 5: 21-28

[11] Chen X et al. Osteoblast-osteoclast interactions. Connect Tissue Res, 2018, 59: 99-107

[12] Keller ET, Brown J. Prostate cancer bone metastases promote both osteolytic and osteoblastic activity. J. Cell Biochem, 2004, 91: 718-729

[13] Guo HF, Vander Kooi CW. Neuropilin functions as an essential cell surface receptor. J. Biol. Chem., 2015, 290: 29120-29126

[14] Roy S et al. Multifaceted role of neuropilins in the immune system: potential targets for immunotherapy. Front. Immunol., 2017, 8: 1228

[15] Yasuoka H et al. Neuropilin-2 expression in breast cancer: correlation with lymph node metastasis, poor prognosis, and regulation of CXCR4 expression. BMC Cancer, 2009, 9: 220

[16] Zhang L et al. VEGF-A/Neuropilin 1 pathway confers cancer stemness via activating wnt/beta-catenin axis in breast cancer cells. Cell Physiol. Biochem., 2017, 44: 1251-1262

[17] Zhu H, Cai H, Tang M, Tang J. Neuropilin-1 is overexpressed in osteosarcoma and contributes to tumor progression and poor prognosis. Clin. Transl. Oncol., 2014, 16: 732-738

[18] Dutta S et al. Neuropilin-2 regulates endosome maturation and EGFR trafficking to support cancer cell pathobiology. Cancer Res., 2016, 76: 418-428

[19] Stanton MJ et al. Autophagy control by the VEGF-C/NRP-2 axis in cancer and its implication for treatment resistance. Cancer Res, 2013, 73: 160-171

[20] Verlinden L et al. Nrp2 deficiency leads to trabecular bone loss and is accompanied by enhanced osteoclast and reduced osteoblast numbers. Bone, 2013, 55: 465-475

[21] Abida W et al. Genomic correlates of clinical outcome in advanced prostate cancer. Proc. Natl Acad. Sci. USA, 2019, 116: 11428-11436

[22] Dutta S et al. NRP2 transcriptionally regulates its downstream effector WDFY1. Sci. Rep., 2016, 6: 23588

[23] Lin DL et al. Bone metastatic LNCaP-derivative C4-2B prostate cancer cell line mineralizes in vitro. Prostate, 2001, 47: 212-221

[24] Zhang J et al. Osteoprotegerin inhibits prostate cancer-induced osteoclastogenesis and prevents prostate tumor growth in the bone. J. Clin. Invest., 2001, 107: 1235-1244

[25] Di Giacomo V et al. DeltaNp63alpha promotes adhesion of metastatic prostate cancer cells to the bone through regulation of CD82. Oncogene, 2017, 36: 4381-4392

[26] Nandana S et al. Bone metastasis of prostate cancer can be therapeutically targeted at the TBX2-WNT signaling axis. Cancer Res., 2017, 77: 1331-1344

[27] Zhao Q, Shao J, Chen W, Li YP. Osteoclast differentiation and gene regulation. Front Biosci., 2007, 12: 2519-2529

[28] Kim JH, Kim N. Regulation of NFATc1 in osteoclast differentiation. J. Bone Metab., 2014, 21: 233-241

[29] Lieben L, Carmeliet G. The involvement of TRP channels in bone homeostasis. Front Endocrinol. (Lausanne), 2012, 3: 99

[30] Prakriya M, Lewis RS. Potentiation and inhibition of Ca²⁺ release-activated Ca²⁺ channels by 2-aminoethyldiphenyl borate (2-APB) occurs independently of IP(3) receptors. J. Physiol., 2001, 536: 3-19

[31] Feng X, Teitelbaum SL. Osteoclasts: New Insights. Bone Res, 2013, 1: 11-26

[32] Lee JH et al. CXCL10 promotes osteolytic bone metastasis by enhancing cancer outgrowth and osteoclastogenesis. Cancer Res., 2012, 72: 3175-3186

[33] Xuan W et al. Osteoclast differentiation gene expression profiling reveals chemokine CCL4 mediates RANKL-induced osteoclast migration and invasion via PI3K pathway. Cell Biochem. Funct., 2017, 35: 171-177

[34] Grassi F et al. CXCL12 chemokine up-regulates bone resorption and MMP-9 release by human osteoclasts: CXCL12 levels are increased in synovial and bone tissue of rheumatoid arthritis patients. J. Cell Physiol., 2004, 199: 244-251

[35] Rao M, Supakorndej T, Schmidt AP, Link DC. Osteoclasts are dispensable for hematopoietic progenitor mobilization by granulocyte colony-stimulating factor in mice. Exp. Hematol., 2015, 43: 110-4 e111-e112

[36] Li S et al. Granulocyte colony-stimulating factor induces osteoblast inhibition by B lymphocytes and osteoclast activation by T lymphocytes during hematopoietic stem/progenitor cell mobilization. Biol. Blood Marrow. Transpl., 2015, 21: 1384-1391

[37] Shiratori T et al. IL-1beta induces pathologically activated osteoclasts bearing extremely high levels of resorbing activity: a possible pathological subpopulation of osteoclasts, accompanied by suppressed expression of Kindlin-3 and Talin-1. J. Immunol., 2018, 200: 218-228

[38] Herrero AB et al. Effects of IL-8 up-regulation on cell survival and osteoclastogenesis in multiple myeloma. Am. J. Pathol., 2016, 186: 2171-2182

[39] Yoshitake F, Itoh S, Narita H, Ishihara K, Ebisu S. Interleukin-6 directly inhibits osteoclast differentiation by suppressing receptor activator of NF-kappaB signaling pathways. J. Biol. Chem., 2008, 283: 11535-11540

[40] Dai SM, Nishioka K, Yudoh K. Interleukin (IL) 18 stimulates osteoclast formation through synovial T cells in rheumatoid arthritis: comparison with IL1 beta and tumour necrosis factor alpha. Ann. Rheum. Dis., 2004, 63: 1379-1386

[41] Lee MS et al. GM-CSF regulates fusion of mononuclear osteoclasts into bone-resorbing osteoclasts by activating the Ras/ERK pathway. J. Immunol., 2009, 183: 3390-3399

[42] Miyamoto T et al. Bifurcation of osteoclasts and dendritic cells from common progenitors. Blood, 2001, 98: 2544-2554

[43] American Cancer Society. Facts & Figures (2020).

[44] Shiozawa Y et al. Human prostate cancer metastases target the hematopoietic stem cell niche to establish footholds in mouse bone marrow. J. Clin. Invest, 2011, 121: 1298-1312

[45] Shiozawa Y, Pienta KJ, Taichman RS. Hematopoietic stem cell niche is a potential therapeutic target for bone metastatic tumors. Clin. Cancer Res, 2011, 17: 5553-5558

[46] Israeli RS. Managing bone loss and bone metastases in prostate cancer patients: a focus on bisphosphonate therapy. Rev. Urol., 2008, 10: 99-110

[47] Kikuta J, Ishii M. Osteoclast migration, differentiation and function: novel therapeutic targets for rheumatic diseases. Rheumatol. (Oxf.), 2013, 52: 226-234

[48] Ikeda K, Takeshita S. The role of osteoclast differentiation and function in skeletal homeostasis. J. Biochem., 2016, 159: 1-8

[49] Novack DV, Teitelbaum SL. The osteoclast: friend or foe? Annu. Rev. Pathol., 2008, 3: 457-484

[50] Park JH, Lee NK, Lee SY. Current understanding of RANK signaling in osteoclast differentiation and maturation. Mol. Cells, 2017, 40: 706-713

[51] Roy S et al. Macrophage-derived neuropilin-2 exhibits novel tumor-promoting functions. Cancer Res, 2018, 78: 5600-5617

Funding

rudolf becker foundation for translational prostate cancer research