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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

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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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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

Author notes

  1. Fengyang Zheng and Pan Li contributed equally to this work.

Authors and Affiliations

  1. Department of Radiology, Molecular Imaging Program at Stanford (MIPS), Stanford University School of Medicine, 3155 Porter Drive, Palo Alto, CA, 94305, USA

    Fengyang Zheng, Pan Li, Sunitha V. Bachawal, Huaijun Wang & Ramasamy Paulmurugan

  2. Department of Ultrasound, Zhongshan Hospital, Fudan University, 180 Fenglin Rd, Shanghai, 200032, People’s Republic of China

    Fengyang Zheng, Chaolun Li & Beijian Huang

  3. Shanghai Institute of Medical Imaging, Shanghai, 200032, People’s Republic of China

    Fengyang Zheng, Chaolun Li & Beijian Huang

  4. Institute of Ultrasound Imaging of Chongqing Medical University, Chongqing, 400010, People’s Republic of China

    Pan Li

  5. Department of Ultrasound, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400010, People’s Republic of China

    Pan Li

  6. Department of pathology, Zhongshan Hospital, Fudan University, 180 Fenglin Rd, Shanghai, 200032, People’s Republic of China

    Wei Yuan

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  1. Fengyang Zheng
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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 22, 1003–1011 (2020). https://doi.org/10.1007/s11307-020-01478-9

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