Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Sep 28:14:1272570.
doi: 10.3389/fimmu.2023.1272570. eCollection 2023.

A novel [89Zr]-anti-PD-1-PET-CT to assess response to PD-1/PD-L1 blockade in lung cancer

Ander Puyalto  1   2   3 , María Rodríguez-Remírez  1   2   3 , Inés López  2   3 , Fabiola Iribarren  1   2 , Jon Ander Simón  2   4   5 , Marga Ecay  4   5 , María Collantes  4   5 , Anna Vilalta-Lacarra  1   2 , Alejandro Francisco-Cruz  6 , Jose Luis Solórzano  7   8 , Sergio Sandiego  9 , Iván Peñuelas  3   4   5 , Alfonso Calvo  2   3   10 , Daniel Ajona  2   3   10 , Ignacio Gil-Bazo  1   2   3   9   10
Affiliations

A novel [89Zr]-anti-PD-1-PET-CT to assess response to PD-1/PD-L1 blockade in lung cancer

Ander Puyalto et al. Front Immunol. .

Abstract

Background: Harnessing the anti-tumor immune system response by targeting the program cell death protein (PD-1) and program cell death ligand protein (PD-L1) axis has been a major breakthrough in non-small cell lung cancer (NSCLC) therapy. Nonetheless, conventional imaging tools cannot accurately assess response in immunotherapy-treated patients. Using a lung cancer syngeneic mouse model responder to immunotherapy, we aimed to demonstrate that [89Zr]-anti-PD-1 immuno-PET is a safe and feasible imaging modality to assess the response to PD-1/PD-L1 blockade in NSCLC.

Materials and methods: A syngeneic mouse model responder to anti-PD-1 therapy was used. Tumor growth and response to PD-1 blockade were monitored by conventional 2-deoxy-2-[18F]fluoro-D-glucose ([18F]-FDG) PET scans. Additionally, tumor lymphocyte infiltration was analyzed by the use of an [89Zr]-labeled anti-PD-1 antibody and measured as 89Zr tumor uptake.

Results: Conventional [18F]-FDG-PET scans failed to detect the antitumor activity exerted by anti-PD-1 therapy. However, [89Zr]-anti-PD-1 uptake was substantially higher in mice that responded to PD-1 blockade. The analysis of tumor-infiltrating immune cell populations and interleukins demonstrated an increased anti-tumor effect elicited by activation of effector immune cells in PD-1-responder mice. Interestingly, a positive correlation between [89Zr]-anti-PD-1 uptake and the proportion of tumor-infiltrating lymphocytes (TILs) was found (Cor = 0.8; p = 0.001).

Conclusion: Our data may support the clinical implementation of immuno-PET as a promising novel imaging tool to predict and assess the response of PD-1/PD-L1 inhibitors in patients with NSCLC.

Keywords: PD-1 inhibition; immuno-PET; inhibitor of differentiation 1; lung adenocarcinoma; pseudoprogression.

PubMed Disclaimer

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision.

Figures

Figure 1
Figure 1
[18F]-FDG does not detect the antitumor effect of PD-1 blockade in LLC tumors. (A) Representative PET images at days 6 and 13 of 1[18F]-FDG uptake in LLC cells (LLC Sc) injected in Id1+/+ (C57BL/6J) or Id1-/- (IDKO) (n = 4) mice groups. (B) Quantification of [18F]-FDG SUVmax representing the fold change between the uptake at days 6 and 13 of the four mice groups described in (A). (C) Representative PET images of [18F]-FDG uptake at days 6 and 13 of Id1 silenced LLC cells (LLC sh-ID1) injected in Id1+/+ (C57BL/6J) or Id1-/- (IDKO) (n = 4) mice groups. (D) Quantification of [18F]-FDG SUVmax representing the fold change between the uptake at days 6 and 13 of the four mice groups described in (C). Error bars denote SD.
Figure 2
Figure 2
[89Zr]-anti-PD-1 allows an accurate evaluation of tumor response to immunotherapy in a lung cancer mouse model. (A) Outline of the mouse model. LLC [sh_Control (Sc) and Id1 silenced cells (sh-ID1)] were subcutaneously injected in Id1 +/+ C57BL/6J (C57) and Id1 -/- C57BL/6J (IDKO) mice, and animals were treated with PBS or an anti-PD-1 mAb (days 7, 10; RPM-14 100mg, intraperitoneally) and with [89Zr]-anti-PD-1 (day 14; 100mg, intraperitoneally) (n=4 mice per group). Tumor volume was measured using a caliper (mm3), using [18F]-FDG-PET scan analyses (days 6 and 13) or using [89Zr]-anti-PD-1 PET scans analysis (days 15, 17 and 20 after LLC inoculation). (B) Representative PET-CT images of 89Zr signal at days 15, 17 and 20 after LLC inoculation. (C) 89Zr signal LLC Sc and sh-ID1 cells injected in C57 and IDKO treated with anti-PD-1 at days 15, 17, and 20 after LLC inoculation. Asterisks denote significance (*p < 0.05, **p < 0.005, ***p < 0.001), and error bars denote SD.
Figure 3
Figure 3
[89Zr]-anti-PD-1 uptake correlates with immune T cell infiltration. (A) Representative IHC images illustrating: Left: CD3+ T cells; Middle: CD8+ T cells; Right: CD4+ T cells; Scale bar: 200μm. (B) Quantification of the relative stained area of: Left: CD3+ T cells; Middle: CD8+ T cells; Right: CD4+ T cells. (C) Correlation between 89Zr uptake at day 20 after LLC inoculation and: Left: CD3+ T cells proportion area (Cor = 0.8); Right: CD8+ T cells proportion area (Cor = 0.8). Asterisks denote significance (*p < 0.05, ***p < 0.001), and error bars denote SD.
Figure 4
Figure 4
Id1 inhibition at the tumor-microenvironment promotes proinflammatory interleukin expression and T cell infiltration. (A) Representative IHC images illustrating CD3+ T cells, CD8+ T cells and Id1+ cells of LLC cells inoculated in C57 and IDKO mice. Scale bar: 200μm. (B) Left: Quantification of proportion of relative stained area of CD3+ T cells. Right: Quantification of the proportion of relative stained area of CD8+ T cells of tumor samples illustrated in (A). (C) Representative images of multiplex immunofluorescence staining panel with nuclei (white), Id1 (red), CD3 (green), CD8 (yellow) of LLC cells inoculated in C57 and IDKO mice. Scale bar: 200μm. (D) Left: Quantification of multiplex immunofluorescence staining of CD3+ T cells. Right: Quantification of multiplex immunofluorescence staining of CD8+ T cells of tumor samples illustrated in (C). (E) Relative mRNA expression levels of Il-1b, Ifn-γ and Tnf-α in LLC tumors in C57 and IDKO mice. Asterisks denote significance (*p < 0.05, ***p < 0.001), and error bars denote SD.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Siegel RL, Miller KD, Wagle NS, Jemal A. Cancer statistics, 2023. CA Cancer J Clin (2023) 73:17–48. doi: 10.3322/caac.21763 - DOI - PubMed
    1. Chaft JE, Rimner A, Weder W, Azzoli CG, Kris MG, Cascone T. Evolution of systemic therapy for stages I-III non-metastatic non-small-cell lung cancer. Nat Rev Clin Oncol (2021) 18:547–57. doi: 10.1038/s41571-021-00501-4 - DOI - PMC - PubMed
    1. Król K, Mazur A, Stachyra-Strawa P, Grzybowska-Szatkowska L. Non-small cell lung cancer treatment with molecularly targeted therapy and concurrent radiotherapy-A review. Int J Mol Sci (2023) 24:5858. doi: 10.3390/ijms24065858 - DOI - PMC - PubMed
    1. Rodríguez-Abreu D, Powell SF, Hochmair MJ, Gadgeel S, Esteban E, Felip E, et al. . Pemetrexed plus platinum with or without pembrolizumab in patients with previously untreated metastatic nonsquamous NSCLC: protocol-specified final analysis from KEYNOTE-189. Ann Oncol (2021) 32:881–95. doi: 10.1016/j.annonc.2021.04.008 - DOI - PubMed
    1. Havel JJ, Chowell D, Chan TA. The evolving landscape of biomarkers for checkpoint inhibitor immunotherapy. Nat Rev Cancer (2019) 19:133–50. doi: 10.1038/s41568-019-0116-x - DOI - PMC - PubMed

Publication types

MeSH terms

Substances

Grants and funding

AP was supported by Fundación Persán and Ayudas predoctorales para la realización de programas de doctorado de interés para Navarra 2021 fellowships. MRR was supported by a donation from the family of José Luis Larrea. DA, and IGB were supported by a grant (RD12/0036/0040) from Red Temática de Investigación Cooperativa en Cáncer, Instituto de Salud Carlos III, Spanish Ministry of Economy and Competitiveness & European Regional Development Fund “Una manera de hacer Europa” (FEDER; PI17/00411). IGB was also supported by two grants from Instituto de Salud Carlos III (PI15/02223 and PI19/00678), two grants from the Gobierno de Navarra cofunded by the Fondo Europeo de Desarrollo Regional 2014-2020 of Navarra (44/2017 and 53/2021). DA was also supported by the Fundación Científica de la Asociación Española Contra el Cáncer (IDEAS211016AJON), Gobierno de Navarra cofunded by the Fondo Europeo de Desarrollo Regional 2014-2020 of Navarra (51-2021), and Fondo de Investigación Sanitaria-Fondo Europeo de Desarrollo Regional Una manera de hacer Europa (PI20/00419). This study has received a grant for medical writing and article editing from Sociedad Española de Oncología (SEOM). The author(s) declare financial support was received for the research, authorship, and/or publication of this article.