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Noninvasive evaluation of PD-L1 expression in non-small cell lung cancer by immunoPET imaging using an acylating agent–modified antibody fragment

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Abstract

Purpose

The aim of this study was to explore an effective 124I labeling strategy and improve the signal-to-noise ratio when evaluating the expression of PD-L1 using an 124I-iodinated durvalumab (durva) F(ab')2 fragment.

Methods

The prepared durva F(ab')2 fragments were incubated with N-succinimidyl-3-(4-hydroxyphenyl) propionate (SHPP); after purification, the HPP-durva F(ab')2 was iodinated using Iodo-Gen method. After the radiochemical purity, stability, and specific activities were determined, the binding affinities of probes prepared using different labeling strategies were compared in vitro. The clinical application value of [124I]I-HPP-durva-F(ab')2 was confirmed by PET imaging. To more objectively evaluate the in vivo distribution and clearance of tracers, the pharmacokinetics and biodistribution assays were also performed.

Results

After being modified with SHPP, the average conjugation number of SHPP per durva-F(ab')2 identified by LC–MS was about 8.92 ± 2.84. The prepared [124I]I-HPP-durva F(ab')2 was obtained with a satisfactory radiochemical purity of more than 98% and stability of more than 93% when incubated for 72 h. Compared with unmodified [124I]I-durva F(ab')2, the specific activity of [124I]I-HPP-durva-F(ab')2 was improved (52.91 ± 5.55 MBq/mg and 15.91 ± 0.74 MBq/mg), while the affinity did not significantly change. The biodistribution experiments and PET imaging showed that the prepared [124I]I-HPP-durva-F(ab')2 exhibited an accelerated clearance and improved tumor-to-background ratio compared with [124I]I-durva-F(ab')2. The specificity of [124I]I-HPP-durva-F(ab')2 to PD-L1 was well demonstrated both in vitro and in vivo.

Conclusions

A PD-L1 PET imaging probe [124I]I-HPP-durva F(ab')2 was successfully synthesized through the SHPP modification strategy. The prepared probe was able to accurately evaluate the PD-L1 expression level through high-contrast noninvasive imaging.

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Data Availability

The data used to support the findings of this study are available from the corresponding authors upon reasonable request.

References

  1. Dermani FK, Samadi P, Rahmani G, Kohlan AK, Najafi R. PD-1/PD-L1 immune checkpoint: potential target for cancer therapy. J Cell Physiol. 2019;234:1313–25. https://doi.org/10.1002/jcp.27172.

    Article  CAS  PubMed  Google Scholar 

  2. Su D, Tsai HI, Xu Z, Yan F, Wu Y, Xiao Y, et al. Exosomal PD-L1 functions as an immunosuppressant to promote wound healing. J Extracell Vesicles. 2019;9:1709262. https://doi.org/10.1080/20013078.2019.1709262.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Wan W, Ao X, Chen Q, Yu Y, Ao L, Xing W, et al. METTL3/IGF2BP3 axis inhibits tumor immune surveillance by upregulating N(6)-methyladenosine modification of PD-L1 mRNA in breast cancer. Mol Cancer. 2022;21:60. https://doi.org/10.1186/s12943-021-01447-y.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Moik F, Chan WE, Wiedemann S, Hoeller C, Tuchmann F, Aretin MB, et al. Incidence, risk factors, and outcomes of venous and arterial thromboembolism in immune checkpoint inhibitor therapy. Blood. 2021;137:1669–78. https://doi.org/10.1182/blood.2020007878.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Carlisle JW, Steuer CE, Owonikoko TK, Saba NF. An update on the immune landscape in lung and head and neck cancers. CA Cancer J Clin. 2020;70:505–17. https://doi.org/10.3322/caac.21630.

    Article  PubMed  Google Scholar 

  6. Wen M, Cao Y, Wu B, Xiao T, Cao R, Wang Q, et al. PD-L1 degradation is regulated by electrostatic membrane association of its cytoplasmic domain. Nat Commun. 2021;12:5106. https://doi.org/10.1038/s41467-021-25416-7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Neubert NJ, Schmittnaegel M, Bordry N, Nassiri S, Wald N, Martignier C, et al. T cell-induced CSF1 promotes melanoma resistance to PD1 blockade. Sci Transl Med. 2018;10. https://doi.org/10.1126/scitranslmed.aan3311.

  8. Doroshow DB, Bhalla S, Beasley MB, Sholl LM, Kerr KM, Gnjatic S, et al. PD-L1 as a biomarker of response to immune-checkpoint inhibitors. Nat Rev Clin Oncol. 2021;18:345–62. https://doi.org/10.1038/s41571-021-00473-5.

    Article  CAS  PubMed  Google Scholar 

  9. Lu S, Stein JE, Rimm DL, Wang DW, Bell JM, Johnson DB, et al. Comparison of biomarker modalities for predicting response to PD-1/PD-L1 checkpoint blockade: a systematic review and meta-analysis. JAMA Oncol. 2019;5:1195–204. https://doi.org/10.1001/jamaoncol.2019.1549.

    Article  PubMed  PubMed Central  Google Scholar 

  10. Niemeijer AN, Leung D, Huisman MC, Bahce I, Hoekstra OS, van Dongen G, et al. Whole body PD-1 and PD-L1 positron emission tomography in patients with non-small-cell lung cancer. Nat Commun. 2018;9:4664. https://doi.org/10.1038/s41467-018-07131-y.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Yan Y, Zheng L, Du Q, Cui X, Dong K, Guo Y, et al. Interferon regulatory factor 1 (IRF-1) downregulates checkpoint kinase 1 (CHK1) through miR-195 to upregulate apoptosis and PD-L1 expression in Hepatocellular carcinoma (HCC) cells. Br J Cancer. 2021;125:101–11. https://doi.org/10.1038/s41416-021-01337-6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Zhang W, Liu Y, Yan Z, Yang H, Sun W, Yao Y, et al. IL-6 promotes PD-L1 expression in monocytes and macrophages by decreasing protein tyrosine phosphatase receptor type O expression in human hepatocellular carcinoma. J Immunother Cancer. 2020;8. https://doi.org/10.1136/jitc-2019-000285.

  13. Siu LL, Even C, Mesia R, Remenar E, Daste A, Delord JP, et al. Safety and efficacy of durvalumab with or without tremelimumab in patients with PD-L1-low/negative recurrent or metastatic HNSCC: the phase 2 CONDOR randomized clinical trial. JAMA Oncol. 2019;5:195–203. https://doi.org/10.1001/jamaoncol.2018.4628.

    Article  PubMed  Google Scholar 

  14. Schofield DJ, Percival-Alwyn J, Rytelewski M, Hood J, Rothstein R, Wetzel L, et al. Activity of murine surrogate antibodies for durvalumab and tremelimumab lacking effector function and the ability to deplete regulatory T cells in mouse models of cancer. MAbs. 2021;13:1857100. https://doi.org/10.1080/19420862.2020.1857100.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Cheng Y, Shi D, Xu Z, Gao Z, Si Z, Zhao Y, et al. 124I-labeled monoclonal antibody and fragment for the noninvasive evaluation of tumor PD-L1 expression in vivo. Mol Pharm. 2022. https://doi.org/10.1021/acs.molpharmaceut.2c00084.

    Article  PubMed  PubMed Central  Google Scholar 

  16. Tijink BM, Perk LR, Budde M, Stigter-van Walsum M, Visser GW, Kloet RW, et al. 124I–L19-SIP for immuno-PET imaging of tumour vasculature and guidance of 131I–L19-SIP radioimmunotherapy. Eur J Nucl Med Mol Imaging. 2009;36:1235–44. https://doi.org/10.1007/s00259-009-1096-y.

    Article  PubMed  PubMed Central  Google Scholar 

  17. Shi D, Zhang Y, Xu Z, Si Z, Cheng Y, Cheng D, et al. Noninvasive evaluation of EGFR expression of digestive tumors using 99mTc-MAG3-Cet-F(ab’)2-based SPECT/CT imaging. Mol Imaging. 2022;2022:3748315. https://doi.org/10.1155/2022/3748315.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. England CG, Ehlerding EB, Hernandez R, Rekoske BT, Graves SA, Sun H, et al. Preclinical pharmacokinetics and biodistribution studies of 89Zr-labeled pembrolizumab. J Nucl Med. 2017;58:162–8. https://doi.org/10.2967/jnumed.116.177857.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Dong W, Wu X, Ma S, Wang Y, Nalin AP, Zhu Z, et al. The mechanism of anti-PD-L1 antibody efficacy against PD-L1-negative tumors identifies NK cells expressing PD-L1 as a cytolytic effector. Cancer Discov. 2019;9:1422–37. https://doi.org/10.1158/2159-8290.CD-18-1259.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Cyriac G, Gandhi L. Emerging biomarkers for immune checkpoint inhibition in lung cancer. Semin Cancer Biol. 2018;52:269–77. https://doi.org/10.1016/j.semcancer.2018.05.006.

    Article  CAS  PubMed  Google Scholar 

  21. Sha D, Jin Z, Budczies J, Kluck K, Stenzinger A, Sinicrope FA. Tumor mutational burden as a predictive biomarker in solid tumors. Cancer Discov. 2020;10:1808–25. https://doi.org/10.1158/2159-8290.CD-20-0522.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Zettlitz KA, Tavare R, Knowles SM, Steward KK, Timmerman JM, Wu AM. ImmunoPET of malignant and normal B cells with 89Zr- and 124I-labeled obinutuzumab antibody fragments reveals differential CD20 internalization in vivo. Clin Cancer Res. 2017;23:7242–52. https://doi.org/10.1158/1078-0432.CCR-17-0855.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Oliveira MC, Correia JDG. Biomedical applications of radioiodinated peptides. Eur J Med Chem. 2019;179:56–77. https://doi.org/10.1016/j.ejmech.2019.06.014.

    Article  CAS  PubMed  Google Scholar 

  24. Sugiura G, Kuhn H, Sauter M, Haberkorn U, Mier W. Radiolabeling strategies for tumor-targeting proteinaceous drugs. Molecules. 2014;19:2135–65. https://doi.org/10.3390/molecules19022135.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Vorobyeva A, Schulga A, Rinne SS, Gunther T, Orlova A, Deyev S, et al. Indirect radioiodination of DARPin G3 using N-succinimidyl-para-iodobenzoate improves the contrast of HER2 molecular imaging. Int J Mol Sci. 2019;20. https://doi.org/10.3390/ijms20123047.

  26. Tolmachev V, Mume E, Sjoberg S, Frejd FY, Orlova A. Influence of valency and labelling chemistry on in vivo targeting using radioiodinated HER2-binding affibody molecules. Eur J Nucl Med Mol Imaging. 2009;36:692–701. https://doi.org/10.1007/s00259-008-1003-y.

    Article  PubMed  Google Scholar 

  27. Li H, Frankenfield AM, Houston R, Sekine S, Hao L. Thiol-cleavable biotin for chemical and enzymatic biotinylation and its application to mitochondrial TurboID proteomics. J Am Soc Mass Spectrom. 2021;32:2358–65. https://doi.org/10.1021/jasms.1c00079.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Wang Y, Liu X, Hnatowich DJ. An improved synthesis of NHS-MAG3 for conjugation and radiolabeling of biomolecules with (99m)Tc at room temperature. Nat Protoc. 2007;2:972–8. https://doi.org/10.1038/nprot.2007.144.

    Article  CAS  PubMed  Google Scholar 

  29. Vorobyeva A, Ssmall es CA, Konovalova E, Guler R, Mitran B, Garousi J, et al. Comparison of tumortargeting properties of directly and indirectly radioiodinated designed ankyrin repeat protein (DARPin) G3 variants for molecular imaging of HER2. Int J Oncol 2019;54:1209–20. https://doi.org/10.3892/ijo.2019.4712.

  30. Mitran B, Guler R, Roche FP, Lindstrom E, Selvaraju RK, Fleetwood F, et al. Radionuclide imaging of VEGFR2 in glioma vasculature using biparatopic affibody conjugate: proof-of-principle in a murine model. Theranostics. 2018;8:4462–76. https://doi.org/10.7150/thno.24395.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Funding

This study was supported by the General Programs of the National Nature Science Foundation of China (Nos. 11875114, 82172002, and 82272058) and the Shanghai Municipal Science and Technology Committee of the Shanghai Outstanding Young Academic Leaders Plan (No. 21XD1423500). Clinical Research Project of Zhongshan Hospital, Fudan University (No. 2020ZSLC20).

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Author notes

  1. Yuan Cheng and Dai Shi contributed equally to this paper.

Authors and Affiliations

  1. Department of Nuclear Medicine, Zhongshan Hospital, Fudan University, No. 180 Fenglin Road, Shanghai, 200032, China

    Yuan Cheng, Dai Shi, Renjie Ye, Wenhui Fu, Pengcheng Ma, Zhan Si, Zhan Xu, Qingyu Lin & Dengfeng Cheng

  2. Department of Hepatic Oncology, Zhongshan Hospital, Fudan University, No. 180 Fenglin Road, Shanghai, 200032, China

    Lixin Li

  3. Institute of Nuclear Medicine, Fudan University, Shanghai, 200032, China

    Qingyu Lin & Dengfeng Cheng

  4. Shanghai Institute of Medical Imaging, Shanghai, 200032, China

    Qingyu Lin & Dengfeng Cheng

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  1. Yuan Cheng
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Correspondence to Qingyu Lin or Dengfeng Cheng.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. Animal care and experimental procedures were conducted in compliance with the Helsinki Declaration and approved by the Ethical Committee of Zhongshan Hospital, Fudan University. The use of NSCLC patients tissue microarray was approved by the ethics committee of Outdo Biotech.

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Cheng, Y., Shi, D., Ye, R. et al. Noninvasive evaluation of PD-L1 expression in non-small cell lung cancer by immunoPET imaging using an acylating agent–modified antibody fragment. Eur J Nucl Med Mol Imaging 50, 1585–1596 (2023). https://doi.org/10.1007/s00259-023-06130-6

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