Preclinical evaluation and pilot clinical study of [18F]AlF-NOTA-FAPI-04 for PET imaging of rheumatoid arthritis,European Journal of Nuclear Medicine and Molecular Imaging

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Preclinical evaluation and pilot clinical study of [18F]AlF-NOTA-FAPI-04 for PET imaging of rheumatoid arthritis
European Journal of Nuclear Medicine and Molecular Imaging ( IF 9.1 ) Pub Date : 2022-06-18 , DOI: 10.1007/s00259-022-05836-3
Luna Ge _{1,

2} , Zheng Fu ₃ , Yuchun Wei ₄ , Dandan Shi ₂ , Yun Geng ₅ , Huancai Fan ₂ , Ruojia Zhang ₂ , Yuang Zhang _{1,

2} , Shufeng Li ₆ , Shijie Wang ₄ , Haojun Shi ₇ , Guanhua Song ₈ , Jihong Pan _{1,

2} , Kai Cheng ₃ , Lin Wang _{1,

2}

Affiliation

Department of Rheumatology and Autoimmunology, The First Affiliated Hospital of Shandong First Medical University, Ji'nan, 250014, Shandong, China.
Biomedical Sciences College & Shandong Medicinal Biotechnology Centre, Shandong First Medical University & Shandong Academy of Medical Sciences; NHC Key Laboratory of Biotechnology Drugs (Shandong Academy of Medical Sciences); Key Lab for Rare & Uncommon Diseases of Shandong Province, Ji'nan, 250117, Shandong, China.
Department of PET/CT Center, Shandong Cancer Hospital and Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Ji'nan, 250117, Shandong, China.
Department of Radiology, Shandong Cancer Hospital and Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Ji'nan, 250117, Shandong, China.
Shandong First Medical University & Shandong Academy of Medical Sciences, Ji'nan, 250117, Shandong, China.
Department of Orthopedic Surgery, The First Affiliated Hospital of Shandong First Medical University, Ji'nan, 250014, Shandong, China.
The Second Clinical Medical College, Henan University of Chinese Medicine, Zhengzhou, China.
Institute of Basic Medicine, Shandong First Medical University & Shandong Academy of Medical Sciences, Ji'nan, 250117, Shandong, China.

Purpose

Fibroblast-like synoviocytes (FLSs) are key effector cells in the inflamed joints of patients with rheumatoid arthritis (RA). Previous studies have suggested that fibroblast activation protein (FAP) is highly expressed in RA-derived FLSs and is a specific marker of activated RA FLSs. In this study, we developed aluminum-[¹⁸F]-labeled 1,4,7-triazacyclononane-N,N′,N″-triacetic acid–conjugated FAP inhibitor 04 ([¹⁸F]AlF-NOTA-FAPI-04) to image RA-FLSs in vitro and arthritic joints in collagen-induced arthritis (CIA) mice and RA patients.

Methods

RA FLSs and NIH3T3 cells transfected with FAP were used to perform in vitro–binding studies. Biodistribution was conducted in normal DBA1 mice. Collagen-induced arthritis (CIA) models with different arthritis scores were subjected to [¹⁸F]AlF-NOTA-FAPI-04 and ¹⁸F-FDG PET imaging. Histological examinations were performed to evaluate FAP expression and Cy3 dye–labeled FAPI-04(Cy3-FAPI-04) uptake. Blocking studies with excess unlabeled FAPI-04 in CIA mice and NIH3T3 xenografts in immunocompromised mice were used to evaluate the binding specificity of [¹⁸F]AlF-NOTA-FAPI-04. Additionally, [¹⁸F]AlF-NOTA-FAPI-04 PET imaging was performed on two RA patients.

Results

The binding of [¹⁸F]AlF-NOTA-FAPI-04 increased significantly in RA FLSs and NIH3T3 cells overexpressing FAP compared to their parental controls (^FAP-GFP-NIH3T3 vs. ^GFP-NIH3T3, 2.40 ± 0.078 vs. 0.297 ± 0.05% AD/10⁵ cells; RA FLSs vs. OA FLSs, 1.54 ± 0.064 vs. 0.343 ± 0.056% AD/10⁵ cells). Compared to ¹⁸F-FDG imaging, [18F]AlF-NOTA-FAPI-04 showed high uptake in inflamed joints in the early stage of arthritis, which was positively correlated with the arthritic scores (Pearson r=0.834, P<0.001). In addition, the binding of [¹⁸F]AlF-NOTA-FAPI-04 to cells with high FAP expression and the uptake of [¹⁸F]AlF-NOTA-FAPI-04 in arthritic joints both could be blocked by excessive unlabeled FAPI-04. Fluorescent staining showed that the intensity of Cy3-FAPI-04 binding to FAP increased accordingly as the expression of FAP protein increased in cells and tissue sections. Furthermore, the uptake of [¹⁸F]AlF-NOTA-FAPI-04 in ^FAP-GFP-NIH3T3 xenografts was significantly higher than that in ^GFP-NIH3T3 xenograft (35.44 ± 4.27 vs 7.92 ± 1.83% ID/mL). Finally, [¹⁸F]AlF-NOTA-FAPI-04 PET/CT imaging in RA patients revealed nonphysiologically high tracer uptake in the synovium of arthritic joints.

Conclusion

[¹⁸F]AlF-NOTA-FAPI-04 is a promising radiotracer for imaging RA FLSs and could potentially complement the current noninvasive diagnostic parameters.

中文翻译：

[18F]AlF-NOTA-FAPI-04 用于类风湿性关节炎 PET 成像的临床前评估和试验性临床研究

目的

成纤维细胞样滑膜细胞 (FLS) 是类风湿性关节炎 (RA) 患者发炎关节中的关键效应细胞。先前的研究表明，成纤维细胞活化蛋白（FAP）在 RA 衍生的 FLS 中高表达，并且是活化的 RA FLS 的特异性标志物。在这项研究中，我们开发了铝-[ ¹⁸ F]-标记的 1,4,7-三氮杂环壬烷-N,N',N″-三乙酸共轭 FAP 抑制剂 04 ([ ¹⁸ F]AlF-NOTA-FAPI-04)在体外成像 RA-FLSs 和胶原性关节炎 (CIA) 小鼠和 RA 患者的关节炎关节。

方法

用 FAP 转染的 RA FLS 和 NIH3T3 细胞用于进行体外结合研究。在正常 DBA1 小鼠中进行生物分布。对具有不同关节炎评分的胶原诱导的关节炎 (CIA) 模型进行 [ ¹⁸ F]AlF-NOTA-FAPI-04 和¹⁸ F-FDG PET 成像。进行组织学检查以评估 FAP 表达和 Cy3 染料标记的 FAPI-04 (Cy3-FAPI-04) 摄取。使用 CIA 小鼠中过量未标记的 FAPI-04 和免疫受损小鼠中的 NIH3T3 异种移植物的阻断研究来评估 [ ¹⁸ F]AlF-NOTA-FAPI-04 的结合特异性。此外，对两名 RA 患者进行了 [ ¹⁸ F]AlF-NOTA-FAPI-04 PET 成像。

结果

与其亲代对照相比，[ ¹⁸ F]AlF-NOTA-FAPI-04 的结合在过度表达 FAP 的 RA FLS 和 NIH3T3 细胞中显着增加（^FAP- GFP- NIH3T3对比^GFP -NIH3T3，2.40 ± 0.078 对比 0.297 ± 0.05% AD/10 ^{5 个}细胞；RA FLSs 与 OA FLSs，1.54 ± 0.064 与 0.343 ± 0.056% AD/10 ^{5 个}细胞）。与¹⁸ F-FDG 成像相比，[18F]AlF-NOTA-FAPI-04 在关节炎早期的炎症关节中表现出高摄取，与关节炎评分呈正相关（Pearson r =0.834，P <0.001）。此外，[ ¹⁸ F]AlF-NOTA-FAPI-04 与具有高 FAP 表达的细胞的结合以及 [^{关节炎关节中的18} F]AlF-NOTA-FAPI-04 都可能被过量未标记的 FAPI-04 阻断。荧光染色显示，随着细胞和组织切片中FAP蛋白表达的增加，Cy3-FAPI-04与FAP的结合强度相应增加。此外，[ ¹⁸ F]AlF-NOTA-FAPI-04 在^FAP- GFP- NIH3T3 异种移植物中的吸收明显高于^GFP -NIH3T3 异种移植物中的吸收（35.44 ± 4.27 对 7.92 ± 1.83% ID/mL）。最后，RA 患者的 [ ¹⁸ F]AlF-NOTA-FAPI-04 PET/CT 成像显示关节炎关节滑膜中的非生理性高示踪剂摄取。