Top

European Journal of Nuclear Medicine and Molecular Imaging

Published in:

04-08-2022 | Metastasis | Original Article

Carbonic anhydrase IX stratifies patient prognosis and identifies nodal status in animal models of nasopharyngeal carcinoma using a targeted imaging strategy

Authors: Wenhui Huang, Kun Wang, Weiyuan Huang, Zicong He, Jingming Zhang, Bin Zhang, Zhiyuan Xiong, Kelly McCabe Gillen, Wenzhe Li, Feng Chen, Xing Yang, Shuixing Zhang, Jie Tian

Published in: European Journal of Nuclear Medicine and Molecular Imaging | Issue 13/2022

Abstract

Purpose

Accurate identification of nodal status enables adequate neck irradiation for nasopharyngeal carcinoma (NPC). However, most conventional techniques are unable to pick up occult metastases, leading to underestimation of tumor extensions. Here we investigate the clinical significance of carbonic anhydrase IX (CAIX) in human NPC samples, and develop a CAIX-targeted imaging strategy to identify occult lymph node metastases (LNMs) and extranodal extension (ENE) in animal studies.

Methods

A total of 211 NPC samples are performed CAIX staining, and clinical outcomes are analyzed. The metastatic murine models are generated by foot pad injection of NPC cells, and a CAIX-targeted imaging agent (CAIX-800) is intravenously administered. We adopt fluorescence molecular tomography and ultrasonography (US)-guided spectroscopic photoacoustic (sPA) imaging to perform in vivo studies. Histological and immunohistochemical characterization are carried out via node-by-node analysis.

Results

For clinical samples, 90.1% (91/101) primary tumors, 73.3% (66/90) metastases, and 100% (20/20) local recurrences are CAIX positive. In metastases group, 84.7% (61/72) nodal metastases and 22.2% (4/18) organ metastases are CAIX positive. CAIX expression in primary tumors is significantly associated with NPC stage and prognosis. For animal studies, CAIX-800-based fluorescence imaging achieves 81.3% sensitivity and 93.8% specificity in detecting occult LNMs in vivo, with a minimum detectable diameter of 1.7 mm. Coupled with CAIX-800, US-guided sPA imaging could not only detect subcapsular deposits of metastatic cancer cells 2 weeks earlier than conventional techniques, but also successfully track pathological ENE.

Conclusion

CAIX remarkably expresses in human NPCs and stratifies patient prognosis. In preclinical studies, CAIX-800-based imaging successfully identifies occult LNMs and tracks early stage of pathological ENE. This attractive method shows potential in clinic, allowing medical workers to longitudinally monitor nodal status and helping to reduce unnecessary nodal biopsy for patients with NPC.

Graphical abstract

The schematic diagram for the study. CAIX, carbonic anhydrase IX; NPC, nasopharyngeal carcinoma; US, ultrasonography; sPA, spectroscopic photoacoustic.

Available only for authorised users

Chen YP, Chan ATC, Le QT, Blanchard P, Sun Y, Ma J. Nasopharyngeal carcinoma. Lancet. 2019;394:64–80.PubMedCrossRef

Tang XR, Li YQ, Liang SB, Jiang W, Liu F, Ge WX, et al. Development and validation of a gene expression-based signature to predict distant metastasis in locoregionally advanced nasopharyngeal carcinoma: a retrospective, multicentre, cohort study. Lancet Oncol. 2018;19:382–93.PubMedCrossRef

Wong KCW, Hui EP, Lo KW, Lam WKJ, Johnson D, Li L, et al. Nasopharyngeal carcinoma: an evolving paradigm. Nat Rev Clin Oncol. 2021;18:679–95.PubMedCrossRef

Pereira ER, Kedrin D, Seano G, Gautier O, Meijer EFJ, Jones D, et al. Lymph node metastases can invade local blood vessels, exit the node, and colonize distant organs in mice. Science. 2018;359:1403–7.PubMedPubMedCentralCrossRef

Ubellacker JM, Tasdogan A, Ramesh V, Shen B, Mitchell EC, Martin-Sandoval MS, et al. Lymph protects metastasizing melanoma cells from ferroptosis. Nature. 2020;585:113–8.PubMedPubMedCentralCrossRef

Lydiatt WM, Patel SG, O’Sullivan B, Brandwein MS, Ridge JA, Migliacci JC, et al. Head and neck cancers-major changes in the American Joint Committee on cancer eighth edition cancer staging manual. CA Cancer J Clin. 2017;67:122–37.PubMedCrossRef

Van den Bosch S, Vogel WV, Raaijmakers CP, Dijkema T, Terhaard CHJ, Al-Mamgani A, et al. Implications of improved diagnostic imaging of small nodal metastases in head and neck cancer: radiotherapy target volume transformation and dose de-escalation. Radiother Oncol. 2018;128:472–8.PubMedCrossRef

McMullen CP, Garneau J, Weimar E, Ali S, Farinhas JM, Yu E, et al. Occult nodal disease and occult extranodal extension in patients with oropharyngeal squamous cell carcinoma undergoing primary transoral robotic surgery with neck dissection. JAMA Otolaryngol Head Neck Surg. 2019;145:701–7.PubMedPubMedCentralCrossRef

Bhattacharya P, Mukherjee R. Lymph node extracapsular extension as a marker of aggressive phenotype: classification, prognosis and associated molecular biomarkers. Eur J Surg Oncol. 2021;47:721–31.PubMedCrossRef

10.

Alavi A, Carlin SD, Werner TJ, Al-Zaghal A. Suboptimal sensitivity and specificity of PET and other gross imaging techniques in assessing lymph node metastasis. Mol Imaging Biol. 2019;21:808–11.PubMedCrossRef

11.

Nishio N, van den Berg NS, van Keulen S, Martin BA, Fakurnejad S, Teraphongphom N, et al. Optical molecular imaging can differentiate metastatic from benign lymph nodes in head and neck cancer. Nat Commun. 2019;10:5044.PubMedPubMedCentralCrossRef

12.

Civantos FJ, Zitsch RP, Schuller DE, Agrawal A, Smith RB, Nason R, et al. Sentinel lymph node biopsy accurately stages the regional lymph nodes for T1–T2 oral squamous cell carcinomas: results of a prospective multi-institutional trial. J Clin Oncol. 2010;28:1395–400.PubMedPubMedCentralCrossRef

13.

Abdel-Halim CN, Rosenberg T, Dyrvig AK, Høilund-Carlsen PF, Sørensen JA, Rohde M, et al. Diagnostic accuracy of imaging modalities in detection of histopathological extranodal extension: a systematic review and meta-analysis. Oral Oncol. 2021;114:105169.PubMedCrossRef

14.

Kann BH, Hicks DF, Payabvash S, Mahajan A, Du J, Gupta V, et al. Multi-institutional validation of deep learning for pretreatment identification of extranodal extension in head and neck squamous cell carcinoma. J Clin Oncol. 2020;38:1304–11.PubMedCrossRef

15.

Rankin EB, Giaccia AJ. Hypoxic control of metastasis. Science. 2016;352:175–80.PubMedPubMedCentralCrossRef

16.

Wilson WR, Hay MP. Targeting hypoxia in cancer therapy. Nat Rev Cancer. 2011;11:393–410.PubMedCrossRef

17.

Lou Y, McDonald PC, Oloumi A, Chia S, Ostlund C, Ahmadi A, et al. Targeting tumor hypoxia: suppression of breast tumor growth and metastasis by novel carbonic anhydrase IX inhibitors. Cancer Res. 2011;71:3364–76.PubMedCrossRef

18.

Tafreshi NK, Bui MM, Bishop K, Lloyd MC, Enkemann SA, Lopez AS, et al. Non-invasive detection of breast cancer lymph node metastasis using carbonic anhydrases IX and XII targeted imaging probes. Clin Cancer Res. 2012;18:207–19.PubMedCrossRef

19.

Koukourakis MI, Bentzen SM, Giatromanolaki A, Wilson GD, Daley FM, Saunders MI, et al. Endogenous markers of two separate hypoxia response pathways (hypoxia inducible factor 2 alpha and carbonic anhydrase 9) are associated with radiotherapy failure in head and neck cancer patients recruited in the CHART randomized trial. J Clin Oncol. 2006;24:727–35.PubMedCrossRef

20.

Hui EP, Chan AT, Pezzella F, Turley H, To KF, Poon TC, et al. Coexpression of hypoxia-inducible factors 1alpha and 2alpha, carbonic anhydrase IX, and vascular endothelial growth factor in nasopharyngeal carcinoma and relationship to survival. Clin Cancer Res. 2002;8:2595–604.PubMed

21.

Huang W, Wang K, An Y, Meng H, Gao Y, Xiong ZY, et al. In vivo three-dimensional evaluation of tumour hypoxia in nasopharyngeal carcinomas using FMT-CT and MSOT. Eur J Nucl Med Mol Imaging. 2020;47:1027–38.PubMedCrossRef

22.

Yu Q, Huang SS, Wu ZY, Zheng JD, Chen XY, Nie LM. Label-free visualization of early cancer hepatic micrometastasis and intraoperative image-guided surgery by photoacoustic imaging. J Nucl Med. 2020;61:1079–85.PubMedPubMedCentralCrossRef

23.

Luke GP, Myers JN, Emelianov SY, Sokolov KV. Sentinel lymph node biopsy revisited: ultrasound-guided photoacoustic detection of micrometastases using molecularly targeted plasmonic nanosensors. Cancer Res. 2014;74:5397–408.PubMedPubMedCentralCrossRef

24.

Nishio N, van den Berg NS, Martin BA, van Keulen S, Fakurnejad S, Rosenthal EL, et al. Photoacoustic molecular imaging for the identification of lymph node metastasis in head and neck cancer using an anti-EGFR antibody-dye conjugate. J Nucl Med. 2021;62:648–55.PubMedPubMedCentralCrossRef

25.

Colevas AD, Yom SS, Pfister DG, Spencer S, Adelstein D, Adkins D, et al. NCCN guidelines insights: head and neck cancers, version 1. 2018. J Natl Compr Canc Netw. 2018;16:479–90.PubMedCrossRef

26.

Zong D, Yin L, Zhong Q, Guo WJ, Xu JH, Jiang N, et al. ZNF488 enhances the invasion and tumorigenesis in nasopharyngeal carcinoma via the wnt signaling pathway involving epithelial mesenchymal transition. Cancer Res Treat. 2016;48:334–44.PubMedCrossRef

27.

Wichert M, Krall N, Decurtins W, Franzini RM, Pretto F, Schneider P, et al. Dual-display of small molecules enables the discovery of ligand pairs and facilitates affinity maturation. Nat Chem. 2015;7:241–9.PubMedCrossRef

28.

Yang X, Minn I, Rowe SP, Banerjee SR, Gorin MA, Brummet M, et al. Imaging of carbonic anhydrase IX with an ¹¹¹In-labeled dual-motif inhibitor. Oncotarget. 2015;6:33733–42.PubMedPubMedCentralCrossRef

29.

Ho FC, Tham IW, Earnest A, Lee KM, Lu JJ. Patterns of regional lymph node metastasis of nasopharyngeal carcinoma: a meta-analysis of clinical evidence. BMC Cancer. 2012;12:98.PubMedPubMedCentralCrossRef

30.

Guo LJ, Liu XM, Hua J, Dai L, Tao Y, Cao HM, et al. Differential detection of metastatic and inflammatory lymph nodes using inflow-based vascular-space-occupancy (iVASO) MR imaging. Magn Reson Imaging. 2022;85:128–32.PubMedCrossRef

31.

Krishnan G, van den Berg NS, Nishio N, Juniper G, Pei J, Zhou Q, et al. Metastatic and sentinel lymph node mapping using intravenously delivered Panitumumab-IRDye800CW. Theranostics. 2021;11:7188–98.PubMedPubMedCentralCrossRef

32.

Morawitz J, Bruckmann NM, Dietzel F, Ullrich T, Bittner AK, Hoffmann O, et al. Determining the axillary nodal status with four current imaging modalities including ¹⁸F-FDG PET/MRI in newly diagnosed breast cancer: a comparative study using histopathology as reference standard. J Nucl Med. 2021;62:1677–83.PubMedCentralCrossRef

33.

Jones D, Wang Z, Chen IX, Zhang S, Banerji R, Lei PJ, et al. Solid stress impairs lymphocyte infiltration into lymph-node metastases. Nat Biomed Eng. 2021;5:1426–36.PubMedCrossRef

34.

Horsman MR, Mortensen LS, Petersen JB, Busk M, Overgaard J. Imaging hypoxia to improve radiotherapy outcome. Nat Rev Clin Oncol. 2012;9:674–87.PubMedCrossRef

35.

Liu JN, Bu W, Shi J. Chemical design and synthesis of functionalized probes for imaging and treating tumor hypoxia. Chem Rev. 2017;117:6160–224.PubMedCrossRef

36.

Zhou H, Shen G, Zhang W, Cai H, Zhou Y, Li L. ¹⁸F-FDG PET/CT for the diagnosis of residual or recurrent nasopharyngeal carcinoma after radiotherapy: a meta-analysis. J Nucl Med. 2016;57:342–7.PubMedCrossRef

37.

Lu X, Yan CH, Yuan M, Wei Y, Hu G, Kang Y. In vivo dynamics and distinct functions of hypoxia in primary tumor growth and organotropic metastasis of breast cancer. Cancer Res. 2010;70:3905–14.PubMedPubMedCentralCrossRef

38.

Achen MG, Stacker SA. Exit stage left: a tumor cell’s journey from lymph node to beyond. Trends Cancer. 2018;4:519–22.PubMedCrossRef

39.

Lee CK, Jeong SH, Jang C, Bae H, Kim YH, Park I, et al. Tumor metastasis to lymph nodes requires YAP-dependent metabolic adaptation. Science. 2019;363:644–9.PubMedCrossRef

40.

Xiao BB, Chen QY, Sun XS, Li JB, Luo DH, Sun R, et al. Low value of whole-body dual-modality [¹⁸F]Fluorodeoxyglucose Positron Emission Tomography/Computed Tomography in primary staging of stage I-II nasopharyngeal carcinoma: a nest case-control study. Eur Radiol. 2021;31:5222–33.PubMedPubMedCentralCrossRef

41.

Lowe VJ, Duan F, Subramaniam RM, Sicks JD, Romanoff J, Bartel T, et al. Multicenter trial of [¹⁸F]Fluorodeoxyglucose Positron Emission Tomography/Computed Tomography staging of head and neck cancer and negative predictive value and surgical impact in the N0 neck: results from ACRIN 6685. J Clin Oncol. 2019;37:1704–12.PubMedPubMedCentralCrossRef

42.

Cazzamalli S, Dal Corso CA, Neri D. Acetazolamide serves as selective delivery vehicle for dipeptide-linked drugs to renal cell carcinoma. Mol Cancer Ther. 2016;15:2926–35.PubMedPubMedCentralCrossRef

43.

Kulterer OC, Pfaff S, Wadsak W, Garstka N, Remzi M, Vraka C, et al. A microdosing study with ^99mTc-PHC-102 for the SPECT/CT imaging of primary and metastatic lesions in renal cell carcinoma patients. J Nucl Med. 2021;62:360–5.PubMedPubMedCentralCrossRef

44.

Mao Y, Wang S, Lydiatt W, Shah JP, Colevas AD, Lee AWM, et al. Unambiguous advanced radiologic extranodal extension determined by MRI predicts worse outcomes in nasopharyngeal carcinoma: Potential improvement for future editions of N category systems. Radiother Oncol. 2021;157:114–21.PubMedCrossRef

45.

Rosenthal EL, Moore LS, Tipirneni K, de Boer E, Stevens TM, Hartman YE, et al. Sensitivity and specificity of cetuximab-IRDye800CW to identify regional metastatic disease in head and neck cancer. Clin Cancer Res. 2017;23:4744–52.PubMedPubMedCentralCrossRef

Title: Carbonic anhydrase IX stratifies patient prognosis and identifies nodal status in animal models of nasopharyngeal carcinoma using a targeted imaging strategy
Authors: Wenhui Huang
Kun Wang
Weiyuan Huang
Zicong He
Jingming Zhang
Bin Zhang
Zhiyuan Xiong
Kelly McCabe Gillen
Wenzhe Li
Feng Chen
Xing Yang
Shuixing Zhang
Jie Tian
Publication date: 04-08-2022
Publisher: Springer Berlin Heidelberg
Keyword: Metastasis
Published in: European Journal of Nuclear Medicine and Molecular Imaging / Issue 13/2022
Print ISSN: 1619-7070
Electronic ISSN: 1619-7089
DOI: https://doi.org/10.1007/s00259-022-05922-6

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Carbonic anhydrase IX stratifies patient prognosis and identifies nodal status in animal models of nasopharyngeal carcinoma using a targeted imaging strategy

Abstract

Purpose

Methods

Results

Conclusion

Graphical abstract

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

Graphical abstract

Please log in to get access to this content

Other articles of this Issue 13/2022

Safety of [177Lu]Lu-NeoB treatment: a preclinical study characterizing absorbed dose and acute, early, and late organ toxicity

Diagnostic yield of simultaneous dynamic contrast-enhanced magnetic resonance perfusion measurements and [18F]FET PET in patients with suspected recurrent anaplastic astrocytoma and glioblastoma

Application of artificial intelligence in nuclear medicine and molecular imaging: a review of current status and future perspectives for clinical translation

Intraprocedural MRI-based dosimetry during transarterial radioembolization of liver tumours with holmium-166 microspheres (EMERITUS-1): a phase I trial towards adaptive, image-controlled treatment delivery

Addition of Y-90 radioembolization increases tumor response and local disease control in hepatocellular carcinoma patients receiving sorafenib

Preclinical evaluation of [18F]FDG-PET as a biomarker of lymphoid tissue disease and inflammation in Zika virus infection