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Preclinical Assessment of the Effectiveness of Magnetic Resonance Molecular Imaging of Extradomain-B Fibronectin for Detection and Characterization of Oral Cancer

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Abstract

Purpose

Oral squamous cell carcinoma (OSCC) has not seen a substantial improvement in patient survival despite therapeutic advances, making accurate detection and characterization of the disease a clinical priority. Here, we aim to demonstrate the effectiveness of magnetic resonance imaging (MRI) with the targeted MRI contrast agent MT218 specific to extradomain-B fibronectin (EDB-FN) in the tumor microenvironment for detection and characterization of aggressive OSCC tumors.

Procedures

EDB-FN expression was evaluated in human normal tongue and OSCC specimens with immunohistochemistry. Invasiveness of human CAL27, HSC3, and SCC4 OSCC cells was analyzed with spheroid formation and transwell assays. EDB-FN expression in the cells was analyzed with semiquantitative real-time PCR, western blotting, and a peptide binding study with confocal microscopy. Contrast-enhanced MRI with MT218 was performed on subcutaneous OSCC mouse models at a dose of 0.04 mmol/kg, using gadoteridol (0.1 mmol/kg) as a control.

Results

Strong EDB-FN expression was observed in human untreated primary and metastatic OSCC, reduced expression in treated OSCC, and little expression in normal tongue tissue. SCC4 and HSC3 cell lines demonstrated high invasive potential with high and moderate-EDB-FN expression, respectively, while CAL27 showed little invasive potential and low-EDB-FN expression. In T1-weighted MRI, MT218 produced differential contrast enhancement in the subcutaneous tumor models in correlation with EDB-FN expression in the cancer cells. Enhancement in the high-EDB-FN tumors was greater with MT218 at 0.04 mmol/kg than gadoteridol at 0.1 mmol/kg.

Conclusions

The results suggest EDB-FN has strong potential as an imageable biomarker for aggressive OSCC. MRMI results demonstrate the effectiveness of MT218 and the potential for differential diagnostic imaging of oral cancer for improving the management of the disease.

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Acknowledgments

ZR Lu is M. Frank Rudy and Margaret Domiter Rudy Professor of Biomedical Engineering. We also wish to thank Adam Kresak and Jenifer Mikulan at the Case Comprehensive Cancer Center for their technical assistance on the project.

Funding

This work was supported in part by the National Institutes of Health grants R01 CA194518, R01211762, and R44 CA199826, and by the Tissue Resources Core Facility of the Comprehensive Cancer Center of Case Western Reserve University and University Hospitals of Cleveland (P30 CA43703).

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Authors and Affiliations

  1. Department of Biomedical Engineering, Case Western Reserve University, Cleveland, OH, 44106, USA

    Ryan C. Hall, Nadia R. Ayat, Peter L. Qiao, Amita M. Vaidya, Dan Ma & Zheng-Rong Lu

  2. Department of Endodontics, Case Western Reserve University, Cleveland, OH, 44106, USA

    Anita Aminoshariae

  3. Department of Oral and Maxillofacial Medicine and Diagnostic Sciences, Case Western Reserve University, Cleveland, OH, 44106, USA

    Ivan Stojanov

  4. Department of Pathology, Case Western Reserve University, Cleveland, OH, 44106, USA

    Ivan Stojanov

  5. Case Comprehensive Cancer Center, Case Western Reserve University, Cleveland, OH, 44106, USA

    Zheng-Rong Lu

  6. Cleveland, USA

    Zheng-Rong Lu

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Contributions

Conceptual design of the research was coordinated by ZRL and RCH. All aspects of experimental execution were done by RCH. NRA and PLQ contributed to MRI data acquisition. NRA and AMV contributed to in vitro and in vivo data interpretation. IS reviewed all histological slides. AA assisted on clinical aspects of oral cancer, and DM assisted regarding MRI technique. The manuscript was prepared by RCH and ZRL and critically reviewed by NRA, PLQ, AMV, DM, AA, and IS.

Corresponding author

Correspondence to Zheng-Rong Lu.

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Conflict of Interest

ZRL is a co-founder of Molecular Theranostics, LLC, which is focused on the commercialization of targeted MRI contrast agents.

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Hall, R.C., Ayat, N.R., Qiao, P.L. et al. Preclinical Assessment of the Effectiveness of Magnetic Resonance Molecular Imaging of Extradomain-B Fibronectin for Detection and Characterization of Oral Cancer. Mol Imaging Biol 22, 1532–1542 (2020). https://doi.org/10.1007/s11307-020-01524-6

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