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Novel Gastrin-Releasing Peptide Receptor Targeted Near-Infrared Fluorescence Dye for Image-Guided Surgery of Prostate Cancer

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Abstract

Purpose

Prostate cancer (PCa), the most widespread male cancer in western countries, is generally eradicated by surgery, especially if localized. However, during surgical procedures, it is not always possible to identify malignant tissues by visual inspection. Among the possible consequences, there is the formation of positive surgical margins, often associated with recurrence. In this work, the gastrin-releasing peptide receptor (GRPR), overexpressed in the prostatic carcinoma and not in healthy tissues or in benign hyperplasia (BPH), is proposed as target molecule to design a novel near-infrared fluorescent (NIRF) probe for image-guided prostatectomy.

Procedures

The NIRF dye Sulfo-Cy5.5 was conjugated to a Bombesin-like peptide (BBN), targeting GRPR. The final product, called BBN-Cy5.5, was characterized and tested in vitro on PC-3, DU145, and LnCAP cell lines, using unconjugated Sulfo-Cy5.5 as control. In vivo biodistribution studies were performed by optical imaging in PC-3 tumor-bearing and healthy mice. Finally, simulation of the surgical protocol was carried out.

Results

BBN-Cy5.5 showed high water solubility and a good relative quantum yield. The ability of the probe to recognize the GRPR, highly expressed in PC-3 cells, was tested both in vitro and in vivo, where a significant tumor accumulation was achieved 24 h post-injection. Furthermore, a distinguishable fluorescent signal was visible in mice bearing PCa, when the surgery was simulated. By contrast, low signal was found in healthy or BPH-affected mice.

Conclusions

This work proposes a new NIRF probe ideal to target GRPR, biomarker of PCa. The promising data obtained suggest that the dye could allow the real-time intraoperative visualization of prostate cancer.

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References

  1. 1.

    Siegel RL, Miller KD, Jemal A (2017) Cancer statistics. CA Cancer J Clin 67:7–30

  2. 2.

    Litwin MS, Tan HJ (2017) The diagnosis and treatment of prostate cancer: a review. JAMA 317:2532–2542

  3. 3.

    Trewartha D, Carter K (2013) Advances in prostate cancer treatment. Nat Rev Drug Discov 12:823

  4. 4.

    Chen L, Li Q, Wang Y et al (2017) Comparison on efficacy of radical prostatectomy versus external beam radiotherapy for the treatment of localized prostate cancer. Oncotarget 8:79854–79863

  5. 5.

    Stitzenberg KB, Wong YN, Nielsen ME et al (2012) Trends in radical prostatectomy: centralization, robotics, and access to urologic cancer care. Cancer 118:54–62

  6. 6.

    Patel VR, Shah K, Palmer KJ et al (2007) Robotic-assisted laparoscopic radical prostatectomy: a report of the current state. Expert Rev Anticancer Ther 7:1269–1278

  7. 7.

    Eastham JA, Kuroiwa K, Ohori M et al (2007) Prognostic significance of location of positive margins in radical prostatectomy specimens. Urology 70:965–969

  8. 8.

    Swindle P, Eastham JA, Ohori M et al (2005) Do margins matter? The prognostic significance of positive surgical margins in radical prostatectomy specimens. J Urol 174:903–907

  9. 9.

    Yossepowitch O, Bjartell A, Eastham JA et al (2009) Positive surgical margins in radical prostatectomy: outlining the problem and its long-term consequences. Eur Urol 55:87–99

  10. 10.

    Yossepowitch O, Briganti A, Eastham JA et al (2014) Positive surgical margins after radical prostatectomy: a systematic review and contemporary update. Eur Urol 65:303–313

  11. 11.

    Park YH, Jeong CW, Lee SE (2013) A comprehensive review of neuroanatomy of the prostate. Prostate Int 1:139–145

  12. 12.

    Kung TA, Waljee JF, Curtin CM et al (2015) Interpositional nerve grafting of the prostatic plexus after radical prostatectomy. Plast Reconstr Surg Glob Open 3:e452

  13. 13.

    Keereweer S, Kerrebijn JD, van Driel PB et al (2011) Optical image-guided surgery--where do we stand? Mol Imaging Biol 13:199–207

  14. 14.

    Ntziachristos V, Yoo JS, van Dam GM (2010) Current concepts and future perspectives on surgical optical imaging in cancer. J Biomed Opt 15:066024

  15. 15.

    Stummer W, Pichlmeier U, Meinel T et al (2006) Fluorescence-guided surgery with 5-aminolevulinic acid for resection of malignant glioma: a randomised controlled multicentre phase III trial. Lancet Oncol 7:392–401

  16. 16.

    van Dam GM, Themelis G, Crane LM et al (2011) Intraoperative tumor-specific fluorescence imaging in ovarian cancer by folate receptor-alpha targeting: first in-human results. Nat Med 17:1315–1319

  17. 17.

    Vahrmeijer AL, Hutteman M, van der Vorst JR et al (2013) Image-guided cancer surgery using near-infrared fluorescence. Nat Rev Clin Oncol 10:507–518

  18. 18.

    Verbeek FP, van der Vorst JR, Tummers QR et al (2014) Near-infrared fluorescence imaging of both colorectal cancer and ureters using a low-dose integrin targeted probe. Ann Surg Oncol 21:S528–S537

  19. 19.

    Handgraaf HJM, Boonstra MC, Prevoo H et al (2017) Real-time near-infrared fluorescence imaging using cRGD-ZW800-1 for intraoperative visualization of multiple cancer types. Oncotarget 8:21054–21066

  20. 20.

    Lee JYK, Pierce JT, Zeh R et al (2017) Intraoperative near-infrared optical contrast can localize brain metastases. World Neurosurg 106:120–130

  21. 21.

    Schaafsma BE, Verbeek FP, Elzevier HW et al (2014) Optimization of sentinel lymph node mapping in bladder cancer using near-infrared fluorescence imaging. J Surg Oncol 110:845–850

  22. 22.

    Hachey KJ, Gilmore DM, Armstrong KW et al (2016) Safety and feasibility of near-infrared image-guided lymphatic mapping of regional lymph nodes in esophageal cancer. J Thorac Cardiovasc Surg 152:546–554

  23. 23.

    Liu J, Huang L, Wang N, Chen P (2017) Indocyanine green detects sentinel lymph nodes in early breast cancer. J Int Med Res 45:514–524

  24. 24.

    Tjalma JJ, Garcia-Allende PB, Hartmans E et al (2016) Molecular fluorescence endoscopy targeting vascular endothelial growth factor a for improved colorectal polyp detection. J Nucl Med 57:480–485

  25. 25.

    Gong L, Ding H, Long NE et al (2018) A 3E8.scFv.Cys-IR800 conjugate targeting TAG-72 in an orthotopic colorectal Cancer model. Mol Imaging Biol 20:47–54

  26. 26.

    van Driel PB, Boonstra MC, Prevoo HA et al (2016) EpCAM as multi-tumour target for near-infrared fluorescence guided surgery. BMC Cancer 16:884

  27. 27.

    Christensen A, Juhl K, Persson M et al (2017) uPAR-targeted optical near-infrared (NIR) fluorescence imaging and PET for image-guided surgery in head and neck cancer: proof-of-concept in orthotopic xenograft model. Oncotarget 8:15407–15419

  28. 28.

    Yuan J, Yi X, Yan F et al (2015) Near infrared fluorescence imaging of prostate cancer using heptamethine carbocyanine dyes. Mol Med Rep 11:821–828

  29. 29.

    Lutje S, Rijpkema M, Franssen GM et al (2014) Dual-modality image-guided surgery of prostate cancer with a radiolabeled fluorescent anti-PSMA monoclonal antibody. J Nucl Med 55:995–1001

  30. 30.

    Sonn GA, Behesnilian AS, Jiang ZK et al (2016) Fluorescent image-guided surgery with an anti-prostate stem cell antigen (PSCA) diabody enables targeted resection of mouse prostate cancer xenografts in real time. Clin Cancer Res 22(6):1403–1412

  31. 31.

    Schroeder RP, van Weerden WM, Krenning EP et al (2011) Gastrin-releasing peptide receptor-based targeting using bombesin analogues is superior to metabolism-based targeting using choline for in vivo imaging of human prostate cancer xenografts. Eur J Nucl Med Mol Imaging 38:1257–1266

  32. 32.

    Prignon A, Nataf V, Provost C et al (2015) 68Ga-AMBA and (18)F-FDG for preclinical PET imaging of breast cancer: effect of tamoxifen treatment on tracer uptake by tumor. Nucl Med Biol 42:92–98

  33. 33.

    Dam JH, Olsen BB, Baun C et al (2016) In vivo evaluation of a bombesin analogue labeled with Ga-68 and co-55/57. Mol Imaging Biol 18:368–376

  34. 34.

    Rybalov M, Ananias HJ, Hoving HD et al (2014) PSMA, EpCAM, VEGF and GRPR as imaging targets in locally recurrent prostate cancer after radiotherapy. Int J Mol Sci 15:6046–6061

  35. 35.

    Beer M, Montani M, Gerhardt J et al (2012) Profiling gastrin-releasing peptide receptor in prostate tissues: clinical implications and molecular correlates. Prostate 72:318–325

  36. 36.

    Lantry LE, Cappelletti E, Maddalena ME et al (2006) 177Lu-AMBA: synthesis and characterization of a selective 177Lu-labeled GRP-R agonist for systemic radiotherapy of prostate cancer. J Nucl Med 47:1144–1152

  37. 37.

    Maddalena ME, Fox J, Chen J et al (2009) 177Lu-AMBA biodistribution, radiotherapeutic efficacy, imaging, and autoradiography in prostate cancer models with low GRP-R expression. J Nucl Med 50:2017–2024

  38. 38.

    Liolios C, Schäfer M, Haberkorn U et al (2016) Novel bispecific PSMA/GRPr targeting radioligands with optimized pharmacokinetics for improved PET imaging of prostate cancer. Bioconjug Chem 27:737–751

  39. 39.

    Pu F, Qiao J, Xue S et al (2015) GRPR-targeted protein contrast agents for molecular imaging of receptor expression in cancers by MRI. Sci Rep 18:16214

  40. 40.

    Salinas CA, Tsodikov A, Ishak-Howard M, Cooney KA (2014) Prostate cancer in young men: an important clinical entity. Nat Rev Urol 11:317–323

  41. 41.

    Laidler P, Dulinska J, Lekka M, Lekki J (2005) Expression of prostate specific membrane antigen in androgen-independent prostate cancer cell line PC-3. Arch Biochem Biophys 435:1–14

  42. 42.

    Bouchelouche K, Choyke PL, Capala J (2010) Prostate specific membrane antigen- a target for imaging and therapy with radionuclides. Discov Med 9:55–61

  43. 43.

    Kularatne SA, Thomas M, Myers CH et al (2019) Evaluation of novel prostate-specific membrane antigen-targeted near-infrared imaging agent for fluorescence-guided surgery of prostate cancer. Clin Cancer Res 25:177–187

  44. 44.

    Matsuoka D, Watanabe H, Shimizu Y et al (2017) Synthesis and evaluation of a novel near-infrared fluorescent probe based on succinimidyl-cys-C(O)-glu that targets prostate-specific membrane antigen for optical imaging. Bioorg Med Chem Lett 27:4876–4880

  45. 45.

    Baranski AC, Schäfer M, Bauder-Wüst U et al (2017) PSMA-11-derived dual-labeled PSMA inhibitors for preoperative PET imaging and precise fluorescence-guided surgery of prostate cancer. J Nucl Med 59:639–645

  46. 46.

    Hermann RM, Djannatian M, Czech N, Nitsche M (2016) Prostate-specific membrane antigen PET/CT: false-positive results due to sarcoidosis? Case Rep Oncol 9:457–463

  47. 47.

    Sasikumar A, Joy A, Nanabala R, Pillai MR (2016) 68Ga-PSMA PET/CT false-positive tracer uptake in Paget disease. Clin Nucl Med 41:e454–e455

  48. 48.

    Gu Z, Thomas G, Yamashiro J et al (2000) Prostate stem cell antigen (PSCA) expression increases with high Gleason score, advanced stage and bone metastasis in prostate cancer. Oncogene 19:1288–1296

  49. 49.

    Tsai WK, Zettlitz KA, Tavaré R et al (2018) Dual-modality immunoPET/fluorescence imaging of prostate cancer with an anti-PSCA cys-minibody. Theranostics 8:5903–5914

  50. 50.

    Zhang M, Kobayashi N, Zettlitz KA et al (2019) Near-infrared-dye labeled anti-prostate stem cell antigen minibody enables real-time fluorescence imaging and targeted surgery in translational mouse models. Clin Cancer Res 25:188–200

  51. 51.

    Cai QY, Yu P, Besch-Williford C et al (2013) Near-infrared fluorescence imaging of gastrin releasing peptide receptor targeting in prostate cancer lymph node metastases. Prostate 73:842–854

  52. 52.

    Shrivastava A, Ding H, Kothandaraman S et al (2014) A high-affinity near-infrared fluorescent probe to target bombesin receptors. Mol Imaging Biol 16:661–669

  53. 53.

    Levine L, Lucci JA, Pazdrak B et al (2003) Bombesin stimulates nuclear factor kappa B activation and expression of proangiogenic factors in prostate cancer cells. Cancer Res 63:3495–3502

  54. 54.

    Nagasaki S, Nakamura Y, Maekawa T et al (2012) Immunohistochemical analysis of gastrin-releasing peptide receptor (GRPR) and possible regulation by estrogen receptor betacx in human prostate carcinoma. Neoplasma 59:224–232

  55. 55.

    Markwalder R, Reubi JC (1999) Gastrin-releasing peptide receptors in the human prostate: relation to neoplastic transformation. Cancer Res 59:1152–1159

  56. 56.

    Lee S, Xie J, Chen X (2010) Peptide-based probes for targeted molecular imaging. Biochemistry 49:1364–1376

  57. 57.

    Katsuno T, Pradhan TK, Ryan RR et al (1999) Pharmacology and cell biology of the bombesin receptor subtype 4 (BB4-R). Biochemistry 38:7307–7320

  58. 58.

    Oliveira-Freitas VL, Thomaz LD, Simoneti LE et al (2015) RC-3095, a selective gastrin-releasing peptide receptor antagonist, does not protect the lungs in an experimental model of lung ischemia-reperfusion injury. Biomed Res Int 2015:496378

  59. 59.

    Ohki-Hamazaki H, Iwabuchi M, Maekawa F (2005) Development and function of bombesin-like peptides and their receptors. Int J Dev Biol 49:293–300

  60. 60.

    Xie B, Stammes MA, van Driel PB et al (2015) Necrosis avid near infrared fluorescent cyanines for imaging cell death and their use to monitor therapeutic efficacy in mouse tumor models. Oncotarget 6:39036–39049

  61. 61.

    Bastian A, Thorpe JE, Disch BC et al (2015) A small molecule with anticancer and antimetastatic activities induces rapid mitochondrial-associated necrosis in breast cancer. J Pharmacol Exp Ther 353:392–404

  62. 62.

    Jung DC, Lee HJ, Seo JW et al (2012) Diffusion-weighted imaging of a prostate cancer xenograft model seen on a 7 tesla animal MR scanner: comparison of ADC values and pathologic findings. Korean J Radiol 13:82–89

  63. 63.

    Zhang H, Desai P, Koike et al (2017) Dual-modality imaging of prostate cancer with a fluorescent and radiogallium-labeled gastrin-releasing peptide receptor antagonist. J Nucl Med 58:29–35

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Acknowledgements

Fondazione Umberto Veronesi is gratefully acknowledged for the support.

Dr. Marina Boido is gratefully acknowledged for the help in acquisition of confocal images.

Funding

The project was carried out thanks to the financial support of AIRC Investigator Grant IG 2013.

Author information

Correspondence to Enzo Terreno.

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The authors declare that they have no conflicts of interest.

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Pagoto, A., Garello, F., Marini, G.M. et al. Novel Gastrin-Releasing Peptide Receptor Targeted Near-Infrared Fluorescence Dye for Image-Guided Surgery of Prostate Cancer. Mol Imaging Biol 22, 85–93 (2020). https://doi.org/10.1007/s11307-019-01354-1

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Key words

  • Image-guided surgery
  • Prostate cancer
  • Bombesin
  • Gastrin-releasing peptide receptor (GRPR)
  • Near-infrared probe
  • Orthotopic mouse model