CD11b-Based Pre-Targeted SPECT/CT Imaging Allows for the Detection of Inflammation in Aortic Aneurysm

Abstract

Purpose

To investigate the feasibility of a pre-targeted imaging strategy based on the cycloaddition between 1,2,4,5-terazine (Tz) and trans-cyclooctene (TCO) for evaluating CD11b expression in inflammatory aortic aneurysm (AA) using single photon emission computed tomography/computed tomography (SPECT/CT).

Methods

C57BL/6J mice were fed β-aminopropionitrile (1 g/kg/day) for 4 weeks to establish AA models. Anti-CD11b-TCO was synthesized and ^99mTc-HYNIC-PEG₁₁-Tz was designed for pre-targeted SPECT/CT. The affinity and specificity of the probe for the inflammatory cell line Raw-264.7 were investigated. Then, anti-CD11b-TCO pre-targeted and ^99mTc-HYNIC-PEG₁₁-Tz based SPECT/CT were performed to detect in vivo inflammation in AA. Finally, ex vivo aortic breast-specific gamma imaging (BSGI), Western blot assays, and immunohistochemical CD11b staining were performed to confirm the in vivo findings of SPECT/CT.

Results

In the AA models, 65.22% (15/23) had aortic lesions, including 43.48% (10/23) AA lesions. The anti-CD11b-TCO presented with a high TCO coupling ratio (7.43), and the ^99mTc-HYNIC-PEG₁₁-Tz showed high radio-purity (>95%), good in vitro stability and a rapid clearance rate. Additionally, anti-CD11b-TCO and ^99mTc-HYNIC-PEG₁₁-Tz presented high click rate (~89%). The in vitro clicked compound, ^99mTc-HYNIC-PEG₁₁-Tz/TCO-anti-CD11b, showed high affinity and specificity for Raw-264.7 cells. ^99mTc-HYNIC-PEG₁₁-Tz/TCO-anti-CD11b pre-targeting SPECT/CT successfully demonstrated inflammatory AA with a high AA-to-background ratio in AA mice, compared to AA mice that were injected with ^99mTc-HYNIC-Tz/TCO-IgG (8.13 versus 3.71, P < 0.001) and control mice injected with ^99mTc-HYNIC-Tz/TCO-anti-CD11b (8.13 versus 3.66, P < 0.001). This result was confirmed by ex vivo BSGI performed immediately after SPECT/CT and immunohistochemical CD11b staining.

Conclusion

SPECT/CT imaging using the anti-CD11b-TCO/Tz-PEG₁₁-HYNIC-^99mTc based pre-targeting imaging strategy allows for the detection of inflammation in progressive AA.

Keywords:

• aortic aneurysm
• inflammation
• SPECT/CT
• CD11b
• macrophage

Data Sharing Statement

The dataset used and/or analysed during the current study is available from the corresponding author on reasonable request.

Acknowledgments

We would like to thank for the technical supports from Prof. Shaoli Song and Dr. Jianping Zhang from Center of Biomedical Imaging, Fudan University, and Shanghai Engineering Research Center of Molecular Imaging Probes.

Disclosure

The authors declare that there is no conflicts of interest regarding this article.

Additional information

Funding

This study was funded by the National Natural Science Foundation of China (grant number: 81701730 and 81901796), the Shanghai “Rising Stars of Medical Talent”–Youth Development Program (grant number: HWJRS2019-72), the Shanghai Sailing Program Supported by Shanghai Science and Technology Commission (grant number: 19YF1408300), and the Shanghai Municipal Key Clinical Specialty Project (grant number: SHSLCZDZK03401).