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Assessment of Metastatic and Reactive Sentinel Lymph Nodes with B7-H3-Targeted Ultrasound Molecular Imaging: A Longitudinal Study in Mouse Models

Abstract

Purpose

To explore the potential of B7-H3-targeted ultrasound molecular imaging (USMI) for longitudinal assessment and differentiation of metastatic and reactive sentinel lymph nodes (SLNs) in mouse models.

Procedures

Metastatic and reactive SLN models were established by injection of 4T1 breast cancer cells and complete Freund’s adjuvant (CFA) respectively to the 4th mammary fat pad of female BALB/c mice. At day 21, 28, and 35 after inoculation, USMI was performed following intravenous injection of B7-H3-targeted microbubbles (MBB7-H3) or IgG-control microbubbles (MBcontrol). All SLNs were histopathologically examined after the last imaging session.

Results

A total of 20 SLNs from tumor-bearing mice (T-SLNs) and five SLNs from CFA-injected mice (C-SLNs) were examined by USMI. Nine T-SLNs were histopathologically positive for metastasis (MT-SLNs). From day 21 to 35, T-SLNs showed a rising trend in MBB7-H3 signal with a steep increase in MT-SLNs at day 35 (213.5 ± 80.8 a.u.) as compared to day 28 (87.6 ± 77.2 a.u., P = 0.002) and day 21 (55.7 ± 35.5 a.u., P < 0.001). At day 35, MT-SLNs had significantly higher MBB7-H3 signal than non-metastatic T-SLNs (NMT-SLNs) (101.9 ± 48.0 a.u., P = 0.001) and C-SLNs (38.5 ± 34.0 a.u., P = 0.001); MBB7-H3 signal was significantly higher than MBcontrol in MT-SLNs (P = 0.001), but not in NMT-SLNs or C-SLNs (both P > 0.05). A significant correlation was detected between MBB7-H3 signal and volume fraction of metastasis in MT-SLNs (r = 0.76, P = 0.017).

Conclusions

B7-H3-targeted USMI allows differentiation of MT-SLNs from NMT-SLNs and C-SLNs in mouse models and has great potential to evaluate tumor burden in SLNs of breast cancer.

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References

  1. 1.

    Gradishar WJ, Anderson BO, Balassanian R et al (2017) NCCN guidelines insights: breast cancer, version 1.2017. J Natl Compr Cancer Netw 15:433–451

  2. 2.

    Banerjee M, George J, Song EY et al (2004) Tree-based model for breast cancer prognostication. J Clin Oncol 22:2567–2575

  3. 3.

    Nemoto T, Vana J, Bedwani RN et al (1980) Management and survival of female breast cancer: results of a national survey by the American College of Surgeons. Cancer 45:2917–2924

  4. 4.

    Fisher B, Bauer M, Wickerham DL et al (1983) Relation of number of positive axillary nodes to the prognosis of patients with primary breast cancer. An NSABP update. Cancer 52:1551–1557

  5. 5.

    Soares EWS, Nagai HM, Bredt LC et al (2014) Morbidity after conventional dissection of axillary lymph nodes in breast cancer patients. World J Surg Oncol 12:67

  6. 6.

    Lucci A, McCall LM, Beitsch PD et al (2007) Surgical complications associated with sentinel lymph node dissection (SLND) plus axillary lymph node dissection compared with SLND alone in the American College of Surgeons Oncology Group Trial Z0011. J Clin Oncol 25:3657–3663

  7. 7.

    Veronesi U, Paganelli G, Viale G et al (2003) A randomized comparison of sentinel-node biopsy with routine axillary dissection in breast cancer. N Engl J Med 349:546–553

  8. 8.

    Ashikaga T, Krag DN, Land SR et al (2010) Morbidity results from the NSABP B-32 trial comparing sentinel lymph node dissection versus axillary dissection. J Surg Oncol 102:111–118

  9. 9.

    Montgomery LL, Thorne AC, Van Zee KJ et al (2002) Isosulfan blue dye reactions during sentinel lymph node mapping for breast cancer. Anesth Analg 95:385–388 table of contents

  10. 10.

    Pesek S, Ashikaga T, Krag LE, Krag D (2012) The false-negative rate of sentinel node biopsy in patients with breast cancer: a meta-analysis. World J Surg 36:2239–2251

  11. 11.

    Krag DN, Anderson SJ, Julian TB et al (2007) Technical outcomes of sentinel-lymph-node resection and conventional axillary-lymph-node dissection in patients with clinically node-negative breast cancer: results from the NSABP B-32 randomised phase III trial. Lancet Oncol 8:881–888

  12. 12.

    Esen G, Gurses B, Yilmaz MH et al (2005) Gray scale and power Doppler US in the preoperative evaluation of axillary metastases in breast cancer patients with no palpable lymph nodes. Eur Radiol 15:1215–1223

  13. 13.

    Esen G (2006) Ultrasound of superficial lymph nodes. Eur J Radiol 58:345–359

  14. 14.

    Alvarez S, Añorbe E, Alcorta P et al (2006) Role of sonography in the diagnosis of axillary lymph node metastases in breast cancer: a systematic review. AJR Am J Roentgenol 186:1342–1348

  15. 15.

    Mori N, Mugikura S, Miyashita M et al (2018) Perfusion contrast-enhanced ultrasound to predict early lymph-node metastasis in breast cancer. Jpn J Radiol 37:145–153

  16. 16.

    Zhao J, Zhang J, Zhu QL et al (2018) The value of contrast-enhanced ultrasound for sentinel lymph node identification and characterisation in pre-operative breast cancer patients: a prospective study. Eur Radiol 28:1654–1661

  17. 17.

    Goldberg BB, Merton DA, Liu J-B et al (2004) Sentinel lymph nodes in a swine model with melanoma: contrast-enhanced lymphatic US. Radiology 230:727–734

  18. 18.

    Wang Y, Cheng Z, Li J, Tang J (2010) Gray-scale contrast-enhanced ultrasonography in detecting sentinel lymph nodes: an animal study. Eur J Radiol 74:e55–e59

  19. 19.

    Goldberg BB, Merton DA, Bin LJ et al (2011) Contrast-enhanced ultrasound imaging of sentinel lymph nodes after peritumoral administration of sonazoid in a melanoma tumor animal model. J Ultrasound Med 30:441–453

  20. 20.

    Poanta L, Serban O, Pascu I et al (2014) The place of CEUS in distinguishing benign from malignant cervical lymph nodes: a prospective study. Med Ultrason 16:7–14

  21. 21.

    Slaisova R, Benda K, Jarkovsky J et al (2013) Contrast-enhanced ultrasonography compared to gray-scale and power doppler in the diagnosis of peripheral lymphadenopathy. Eur J Radiol 82:693–698

  22. 22.

    Hong YR, Luo ZY, Mo GQ et al (2017) Role of contrast-enhanced ultrasound in the pre-operative diagnosis of cervical lymph node metastasis in patients with papillary thyroid carcinoma. Ultrasound Med Biol 43:2567–2575

  23. 23.

    Rubaltelli L, Corradin S, Dorigo A et al (2007) Automated quantitative evaluation of lymph node perfusion on contrast-enhanced sonography. Am J Roentgenol 188:977–983

  24. 24.

    Kiessling F, Bzyl J, Fokong S et al (2012) Targeted ultrasound imaging of cancer: an emerging technology on its way to clinics. Curr Pharm Des 18:2184–2199

  25. 25.

    Abou-Elkacem L, Bachawal SV, Willmann JK (2015) Ultrasound molecular imaging: moving toward clinical translation. Eur J Radiol 84:1685–1693

  26. 26.

    Kiessling F, Fokong S, Bzyl J et al (2014) Recent advances in molecular, multimodal and theranostic ultrasound imaging. Adv Drug Deliv Rev 72:15–27

  27. 27.

    Seaman S, Stevens J, Yang MY et al (2007) Genes that distinguish physiological and pathological angiogenesis. Cancer Cell 11:539–554

  28. 28.

    Bachawal SV, Jensen KC, Wilson KE et al (2015) Breast cancer detection by B7-H3-targeted ultrasound molecular imaging. Cancer Res 75:2501–2509

  29. 29.

    Turtoi A, Dumont B, Greffe Y et al (2011) Novel comprehensive approach for accessible biomarker identification and absolute quantification from precious human tissues. J Proteome Res 10:3160–3182

  30. 30.

    Arigami T, Narita N, Mizuno R et al (2010) B7-H3 ligand expression by primary breast cancer and associated with regional nodal metastasis. Ann Surg 252:1044–1051

  31. 31.

    Liu C, Liu J, Wang J et al (2013) B7-H3 expression in ductal and lobular breast cancer and its association with IL-10. Mol Med Rep 7:134–138

  32. 32.

    Nam K, Stanczak M, Forsberg F et al (2018) Sentinel lymph node characterization with a dual-targeted molecular ultrasound contrast agent. Mol Imaging Biol 20:221–229

  33. 33.

    Weiss LM, O’Malley D (2013) Benign lymphadenopathies. Mod Pathol 26(Suppl 1):S88–S96

  34. 34.

    Lei J, Xue HD, Li Z et al (2010) Possible pathological basis for false diagnoses of lymph nodes by USPIO-enhanced MRI in rabbits. J Magn Reson Imaging 31:1428–1434

  35. 35.

    Paschall AV, Liu K (2016) An orthotopic mouse model of spontaneous breast cancer metastasis. J Vis Exp 114, e54040. https://doi.org/10.3791/54040

  36. 36.

    Tafreshi NK, Enkemann SA, Bui MM et al (2011) A mammaglobin-a targeting agent for noninvasive detection of breast cancer metastasis in lymph nodes. Cancer Res 71:1050–1059

  37. 37.

    Willmann JK, Paulmurugan R, Chen K et al (2008) US imaging of tumor angiogenesis with microbubbles targeted to vascular endothelial growth factor receptor type 2 in mice. Radiology 246:508–518

  38. 38.

    Ley K, Laudanna C, Cybulsky MI, Nourshargh S (2007) Getting to the site of inflammation: the leukocyte adhesion cascade updated. Nat Rev Immunol 7:678–689

  39. 39.

    Jin X, Liang N, Wang M et al (2016) Integrin imaging with 99mTc-3PRGD2 SPECT/CT shows high specificity in the diagnosis of lymph node metastasis from non-small cell lung cancer. Radiology 281:958–966

  40. 40.

    Teicher BA (2006) Tumor models for efficacy determination. Mol Cancer Ther 5:2435–2443

  41. 41.

    Miller F, Care A (2000) Mouse 4T1 breast tumor model. Curr Protoc Immunol 20:1–16

  42. 42.

    Taub RN, Krantz AR, Dresser DW (1970) The effect of localized injection of adjuvant material on the draining lymph node. I Histology. Immunology 18:171–186

  43. 43.

    Krzystyniak K, Kozlowska E, Desjardins R et al (1995) Different T-cell activation by streptozotocin and Freund’s adjuvant in popliteal lymph node (PLN). Int J Immunopharmacol 17:189–196

  44. 44.

    Jiménez-González M, Plaza-García S, Arizeta J et al (2017) A longitudinal MRI study on lymph nodes histiocytosis of a xenograft cancer model. PLoS One 12:1–16

  45. 45.

    Herman PG, Kim CS, de Sousa MA, Mellins HZ (1976) Microcirculation of the lymph node with metastases. Am J Pathol 85:333–348

  46. 46.

    Li C, Torres VC, Tichauer KM (2018) Noninvasive detection of cancer spread to lymph nodes: a review of molecular imaging principles and protocols. J Surg Oncol 118:301–314

  47. 47.

    Li G, Quan Y, Che F, Wang L (2018) B7-H3 in tumors: friend or foe for tumor immunity? Cancer Chemother Pharmacol 81:245–253

  48. 48.

    Sun J, Guo Y-D, Li X-N et al (2014) B7-H3 expression in breast cancer and upregulation of VEGF through gene silence. Onco Targets Ther 7:1979–1986

  49. 49.

    Giuliano AE, Ballman KV, McCall L et al (2017) Effect of axillary dissection vs no axillary dissection on 10-year overall survival among women with invasive breast cancer and sentinel node metastasis. JAMA 318:918

  50. 50.

    Kramer K, Kushner BH, Modak S et al (2010) Compartmental intrathecal radioimmunotherapy: results for treatment for metastatic CNS neuroblastoma. J Neuro-Oncol 97:409–418

  51. 51.

    Loos M, Hedderich DM, Friess H, Kleeff J (2010) B7-h3 and its role in antitumor immunity. Clin Dev Immunol 2010:683875

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Acknowledgments

We thank the Canary Center at Stanford, Department of Radiology for facility and resources. We also thank SCi3 Small Animal Imaging Service Center, Stanford University School of Medicine for providing imaging facilities and data analysis support. We also acknowledge Dr. José G. Vilches-Moure, Veterinary pathologist, Animal Histology Services (AHS) for his advice regarding histological analysis of tissues.

Funding

This research was partially supported by NIH R01CA209888 (RP), NIH R21EB022298 (RP) and The Teal Foundation.

Author information

Correspondence to Beijian Huang or Ramasamy Paulmurugan.

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The authors declare that they have no conflict of interests.

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Zheng, F., Li, P., Bachawal, S.V. et al. Assessment of Metastatic and Reactive Sentinel Lymph Nodes with B7-H3-Targeted Ultrasound Molecular Imaging: A Longitudinal Study in Mouse Models. Mol Imaging Biol (2020). https://doi.org/10.1007/s11307-020-01478-9

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Key Words

  • Ultrasound imaging
  • Molecular imaging
  • Sentinel lymph node
  • Metastasis