Skip to main content

Advertisement

SpringerLink
Log in

Development of novel peptide-based radiotracers for detecting PD-L1 expression and guiding cancer immunotherapy

  • Original Article
  • Published:
European Journal of Nuclear Medicine and Molecular Imaging Aims and scope Submit manuscript

Abstract

Purpose

Due to tumor heterogeneity, immunohistochemistry (IHC) showed poor accuracy in detecting the expression of programmed cell death ligand-1 (PD-L1) in patients. Positron emission tomography (PET) imaging is considered as a non-invasive technique to detect PD-L1 expression at the molecular level visually, real-timely and quantitatively. This study aimed to develop novel peptide-based radiotracers [68Ga]/[18F]AlF-NOTA-IMB for accurately detecting the PD-L1 expression and guiding the cancer immunotherapy.

Methods

NOTA-IMB was prepared by connecting 2,2′-(7-(2-((2,5-dioxopyrrolidin-1-yl)oxy)- 2-oxoethyl)-1,4,7-triazonane-1,4-diyl) diacetic acid (NOTA-NHS) with PD-L1-targeted peptide IMB, and further radiolabeled with 68Ga or 18F-AlF. In vitro binding assay was conducted to confirm the ability of [68Ga]/[18F]AlF-NOTA-IMB to detect the expression of PD-L1. In vivo PET imaging of [68Ga]NOTA-IMB and [18F]AlF-NOTA-IMB in different tumor-bearing mice was performed, and dynamic changes of PD-L1 expression level induced by immunotherapy were monitored. Radioautography, western blotting, immunofluorescence staining and biodistribution analysis were carried out to further evaluate the specificity of radiotracers and efficacy of PD-L1 antibody immunotherapy.

Results

[68Ga]NOTA-IMB and [18F]AlF-NOTA-IMB were both successfully prepared with high radiochemical yield (> 95% and > 60%, n = 5) and radiochemical purity (> 95% and > 98%, n = 5). Both tracers showed high affinity to human and murine PD-L1 with the dissociation constant (Kd) of 1.00 ± 0.16/1.09 ± 0.21 nM (A375-hPD-L1, n = 3) and 1.56 ± 0.58/1.21 ± 0.39 nM (MC38, n = 3), respectively. In vitro cell uptake assay revealed that both tracers can specifically bind to PD-L1 positive cancer cells A375-hPD-L1 and MC38 (5.45 ± 0.33/3.65 ± 0.15%AD and 5.87 ± 0.27/2.78 ± 0.08%AD at 120 min, n = 3). In vivo PET imaging and biodistribution analysis showed that the tracer [68Ga]NOTA-IMB and [18F]AlF-NOTA-IMB had high accumulation in A375-hPD-L1 and MC38 tumors, but low uptake in A375 tumor. Treatment of Atezolizumab induced dynamic changes of PD-L1 expression in MC38 tumor-bearing mice, and the tumor uptake of [68Ga]NOTA-IMB decreased from 3.30 ± 0.29%ID/mL to 1.58 ± 0.29%ID/mL (n = 3, P = 0.026) after five treatments. Similarly, the tumor uptake of [18F]AlF-NOTA-IMB decreased from 3.27 ± 0.63%ID/mL to 0.89 ± 0.18%ID/mL (n = 3, P = 0.0004) after five treatments. However, no significant difference was observed in the tumor uptake before and after PBS treatment. Biodistribution, radioautography, western blotting and immunofluorescence staining analysis further demonstrated that the expression level of PD-L1 in tumor-bearing mice treated with Atezolizumab significantly reduced about 3 times and correlated well with the PET imaging results.

Conclusion

[68Ga]NOTA-IMB and [18F]AlF-NOTA-IMB were successfully prepared for PET imaging the PD-L1 expression noninvasively and quantitatively. Dynamic changes of PD-L1 expression caused by immunotherapy can be sensitively detected by both tracers. Hence, the peptide-based radiotracers [68Ga]NOTA-IMB and [18F]AlF-NOTA-IMB can be applied for accurately detecting the PD-L1 expression in different tumors and monitoring the efficacy of immunotherapy.

Graphical Abstract

Two novel peptide-based radiotracers [68Ga]NOTA-IMB and [18F]AlF-NOTA-IMB were developed as promising agents for evaluating the immunotherapy effect by monitoring the changes of PD-L1 expression in real time.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Scheme 1
Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

Data Availability

All data generated or analyzed during this study are included in this published article and its supplementary information files.

References

  1. Zhang Y, Zhang Z. The history and advances in cancer immunotherapy: understanding the characteristics of tumor-infiltrating immune cells and their therapeutic implications. Cell Mol Immunol. 2020;17:807–21.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Zhu S, Zhang T, Zheng L, Liu H, Song W, Liu D, et al. Combination strategies to maximize the benefits of cancer immunotherapy. J Hematol Oncol. 2021;14:156.

    Article  PubMed  PubMed Central  Google Scholar 

  3. Monk BJ, Enomoto T, Kast WM, McCormack M, Tan DSP, Wu X, et al. Integration of immunotherapy into treatment of cervical cancer: recent data and ongoing trials. Cancer Treat Rev. 2022;106:102385.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Vesely MD, Zhang T, Chen L. Resistance mechanisms to anti-PD cancer immunotherapy. Annu Rev Immunol. 2022;40:45–74.

    Article  CAS  PubMed  Google Scholar 

  5. Patsoukis N, Wang Q, Strauss L, Boussiotis VA. Revisiting the PD-1 pathway. Sci Adv. 2020;6:eabd2712.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Mittendorf EA, Philips AV, Meric-Bernstam F, Qiao N, Wu Y, Harrington S, et al. PD-L1 expression in triple-negative breast cancer. Cancer Immunol Res. 2014;2:361–70.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Ricciuti B, Wang X, Alessi JV, Rizvi H, Mahadevan NR, Li YY, et al. Association of high tumor mutation burden in non-small cell lung cancers with increased immune infiltration and improved clinical outcomes of PD-L1 blockade across PD-L1 expression levels. JAMA Oncol. 2022;8:1160–8.

    Article  PubMed  PubMed Central  Google Scholar 

  8. Chen X, Wang K, Jiang S, Sun H, Che X, Zhang M, et al. eEF2K promotes PD-L1 stabilization through inactivating GSK3β in melanoma. J Immunother Cancer. 2022;10:e004026.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Ri MH, Ma J, Jin X. Development of natural products for anti-PD-1/PD-L1 immunotherapy against cancer. J Ethnopharmacol. 2021;281:114370.

    Article  CAS  PubMed  Google Scholar 

  10. Yi M, Zheng X, Niu M, Zhu S, Ge H, Wu K. Combination strategies with PD-1/PD-L1 blockade: current advances and future directions. Mol Cancer. 2022;21:28.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Shen X, Zhao B. Efficacy of PD-1 or PD-L1 inhibitors and PD-L1 expression status in cancer: meta-analysis. BMJ. 2018;362:k3529.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Yi M, Niu M, Xu L, Luo S, Wu K. Regulation of PD-L1 expression in the tumor microenvironment. J Hematol Oncol. 2021;14:10.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Monaco L, De Bernardi E, Bono F. The, “digital biopsy” in non-small cell lung cancer (NSCLC): a pilot study to predict the PD-L1 status from radiomics features of [18F]FDG PET/CT. Eur J Nucl Med Mol Imaging. 2022;49:3401–11.

    Article  CAS  PubMed  Google Scholar 

  14. Lantuejoul S, Sound-Tsao M, Cooper WA, Girard N, Hirsch FR, Roden AC, et al. PD-L1 testing for lung cancer in 2019: perspective from the IASLC pathology committee. J Thorac Oncol. 2020;15:499–519.

    Article  CAS  PubMed  Google Scholar 

  15. Nimmagadda S. Quantifying PD-L1 expression to monitor immune checkpoint therapy: opportunities and challenges. Cancers. 2020;12:3173.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Yeong J, Lum HYJ, Teo CB, Tan BKJ, Chan YH, Tay RYK, et al. Choice of PD-L1 immunohistochemistry assay influences clinical eligibility for gastric cancer immunotherapy. Gastric Cancer. 2022;25:741–50.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Yoon JY, Nayyar R, Quest G, Pabedinskas D, Pal P, Tsao MS, et al. PD-L1 lineage-specific quantification in malignant pleural effusions of lung adenocarcinoma by flow cytometry. Lung Cancer. 2020;148:55–61.

    Article  PubMed  Google Scholar 

  18. Christensen C, Kristensen LK, Alfsen MZ, Nielsen CH, Kjaer A. Quantitative PET imaging of PD-L1 expression in xenograft and syngeneic tumour models using a site-specifically labelled PD-L1 antibody. Eur J Nucl Med Mol Imaging. 2020;47:1302–13.

    Article  CAS  PubMed  Google Scholar 

  19. Zhou H, Bao G, Wang Z, Zhang B, Li D, Chen L, et al. PET imaging of an optimized anti-PD-L1 probe 68Ga-NODAGA-BMS986192 in immunocompetent mice and non-human primates. EJNMMI Res. 2022;12:35.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Zhou M, Wang X, Chen B, Xiang S, Rao W, Zhang Z, et al. Preclinical and first-in-human evaluation of 18F-labeled D-peptide antagonist for PD-L1 status imaging with PET. Eur J Nucl Med Mol Imaging. 2022;49:4312–24.

    Article  CAS  PubMed  Google Scholar 

  21. Li D, Wang F, Jiang J, Hou X, Ding J, Wang Z, et al. Construction of an iodine-labeled CS1001 antibody for targeting PD-L1 detection and comparison with low-molecular-peptide micro-PET imaging. Mol Pharm. 2022;19:4382–9.

    Article  CAS  PubMed  Google Scholar 

  22. Lv G, Sun X, Qiu L, Sun Y, Li K, Liu Q, et al. PET imaging of tumor PD-L1 expression with a highly specific nonblocking single-domain antibody. J Nucl Med. 2020;61:117–22.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Chen Y, Zhu S, Fu J, Lin J, Sun Y, Lv G, et al. Development of a radiolabeled site-specific single-domain antibody positron emission tomography probe for monitoring PD-L1 expression in cancer. J Pharm Anal. 2022;12:869–78.

    Article  PubMed  PubMed Central  Google Scholar 

  24. Liu Q, Jiang L, Li K, Li H, Lv G, Lin J, et al. Immuno-PET imaging of 68Ga-labeled nanobody Nb109 for dynamic monitoring the PD-L1 expression in cancers. Cancer Immunol Immunoth. 2021;70:1721–33.

    Article  CAS  Google Scholar 

  25. Wen X, Zeng X, Cheng X, Zeng X, Liu J, Zhang Y, et al. PD-L1-targeted radionuclide therapy combined with αPD-L1 antibody immunotherapy synergistically improves the antitumor effect. Mol Pharm. 2022;19:3612–22.

    Article  CAS  PubMed  Google Scholar 

  26. 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;19:3551–62.

    Article  CAS  PubMed  Google Scholar 

  27. Moroz A, Lee CY, Wang YH, Hsiao JC, Sevillano N, Truillet C, et al. A preclinical assessment of 89Zr-atezolizumab identifies a requirement for carrier added formulations not observed with 89Zr-C4. Bioconjug Chem. 2018;29:3476–82.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Jung KH, Park JW, Lee JH, Moon SH, Cho YS, Lee KH. 89Zr-labeled anti-PD-L1 antibody PET monitors gemcitabine therapy-induced modulation of tumor PD-L1 expression. J Nucl Med. 2021;62:656–64.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Cheng Y, Shi D, Ye R, Fu W, Ma P, Si Z, 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. 2023;50:1585–96.

    Article  CAS  PubMed  Google Scholar 

  30. Hu K, Kuan H, Hanyu M, Masayuki H, Xie L, Zhang Y, et al. Developing native peptide-based radiotracers for PD-L1 PET imaging and improving imaging contrast by pegylation. Chem Commun. 2019;55:4162–5.

    Article  Google Scholar 

  31. De Silva RA, Kumar D, Lisok A, Chatterjee S, Wharram B, Venkateswara Rao K, et al. Peptide-based 68Ga-PET radiotracer for imaging PD-L1 expression in cancer. Mol Pharm. 2018;15:3946–52.

    Article  PubMed  PubMed Central  Google Scholar 

  32. Zhou X, Jiang J, Yang X, Liu T, Ding J, Nimmagadda S, et al. First-in-humans evaluation of a PD-L1-binding peptide PET radiotracer in non-small cell lung cancer patients. J Nucl Med. 2022;63:536–42.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Kumar D, Mishra A, Lisok A, Kureshi R, Shelake S, Plyku D, et al. Pharmacodynamic measures within tumors expose differential activity of PD(L)-1 antibody therapeutics. Proc Natl Acad Sci U S A. 2021;118:e2107982118.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Zou S, Liu J, Sun Z, Feng X, Wang Z, Jin Y, et al. Discovery of hPRDX5-based peptide inhibitors blocking PD-1/PD-L1 interaction through in silico proteolysis and rational design. Cancer Chemotherapy Pharmacol. 2020;85:185–93.

    Article  CAS  Google Scholar 

  35. Pauwels E, Cleeren F, Tshibangu T, Koole M, Serdons K, Boeckxstaens L, et al. 18F-AlF-NOTA-octreotide outperforms 68Ga-DOTATATE/NOC PET in neuroendocrine tumor patients: results from a prospective. Multicenter Study J Nucl Med. 2023;64:632–8.

    Article  CAS  PubMed  Google Scholar 

  36. Yamaguchi H, Hsu JM, Yang WH, Hung MC. Mechanisms regulating PD-L1 expression in cancers and associated opportunities for novel small-molecule therapeutics. Nat Rev Clin Oncol. 2022;19:287–305.

    Article  CAS  PubMed  Google Scholar 

  37. Luo H, Yang C, Kuang D, Shi S, Chan AW. Visualizing dynamic changes in PD-L1 expression in non-small cell lung carcinoma with radiolabeled recombinant human PD-1. Eur J Nucl Med Mol Imaging. 2022;49:2735–45.

    Article  CAS  PubMed  Google Scholar 

  38. Chen J, Chen R, Huang S, Zu B, Zhang S. Atezolizumab alleviates the immunosuppression induced by PD-L1-positive neutrophils and improves the survival of mice during sepsis. Mol Med Rep. 2021;23:144.

    Article  CAS  PubMed  Google Scholar 

  39. Herbst RS, Giaccone G, de Marinis F, Reinmuth N, Vergnenegre A, Barrios CH, et al. Atezolizumab for first-line treatment of PD-L1-selected patients with NSCLC. N Engl J Med. 2020;383:1328–39.

    Article  CAS  PubMed  Google Scholar 

  40. Deberle L, Benešová M, Umbricht C, Borgna F, Büchler M, Zhernosekov K, et al. Development of a new class of PSMA radioligands comprising ibuprofen as an albumin-binding entity. Theranostics. 2020;10(4):1678–93.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Funding

This study was financially supported by National Natural Science Foundation of China (22076069), Natural Science Foundation of Jiangsu Province (BK20201135), the Major Scientific Research Project of Jiangsu Commission of Health (ZDA2020007, ZD2022036) and the Science Technology and Development Project of Wuxi (Y20212013). We also acknowledge the financial support received from Jiangsu Provincial Medical Key Laboratory (Key Laboratory of Nuclear Medicine).

Author information

Authors and Affiliations

  1. NHC Key Laboratory of Nuclear Medicine, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, 214063, China

    Shiyu Zhu, Beibei Liang, Yuxuan Zhou, Yinfei Chen, Jiayu Fu, Ling Qiu & Jianguo Lin

  2. Department of Radiopharmaceuticals, School of Pharmacy, Nanjing Medical University, Nanjing, 211166, China

    Shiyu Zhu, Yuxuan Zhou, Yinfei Chen, Jiayu Fu, Ling Qiu & Jianguo Lin

Authors
  1. Shiyu Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Beibei Liang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yuxuan Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yinfei Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jiayu Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ling Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jianguo Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

SZ: experiments, data analysis, manuscript writing; BL: experiments, data analysis and manuscript writing; YZ: experiments and data analysis; YC: experiments and data analysis; JF: experiments and data analysis; LQ and JL: design, supervision, manuscript writing and editing. All authors reviewed and approved the final version.

Corresponding authors

Correspondence to Ling Qiu or Jianguo Lin.

Ethics declarations

Ethics approval

All procedures involving animal studies were reviewed and approved by the Animal Care and Ethics Committee of Jiangsu Institute of Nuclear Medicine (JSINM-2022–014).

Competing interests

The authors declare no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 5406 KB)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhu, S., Liang, B., Zhou, Y. et al. Development of novel peptide-based radiotracers for detecting PD-L1 expression and guiding cancer immunotherapy. Eur J Nucl Med Mol Imaging 51, 625–640 (2024). https://doi.org/10.1007/s00259-023-06480-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00259-023-06480-1

Keywords

Search

Navigation