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Development of Novel ImmunoPET Tracers to Image Human PD-1 Checkpoint Expression on Tumor-Infiltrating Lymphocytes in a Humanized Mouse Model

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Abstract

Purpose

It is well known that cancers exploit immune checkpoints (programmed death 1 receptor (PD-1) and its ligand (PD-L1)) to evade anti-tumor immune responses. Although immune checkpoint (IC) blockade is a promising approach, not all patients respond. Hence, imaging of tumor-infiltrating lymphocytes (TILs) is of high specific interest, as they are known to express PD-1 during activation and subsequent exhaustion in the tumor microenvironment and are thought to be potentially predictive of therapeutic responses to IC blockade.

Procedures

We developed immune-tracers for positron emission tomography (PET) to image hPD-1 status of human peripheral blood mononuclear cells (hPBMCs) adoptively transferred to NOD-scid IL-2Rγnull (NSG) mice (hNSG) bearing A375 human skin melanoma tumors. The anti-PD-1 human antibody (IgG; keytruda) was labeled with either Zr-89 or Cu-64 radiometals to image PD-1-expressing human TILs in vivo.

Results

[89Zr] Keytruda (groups = 2; NSG-ctl (control) and hNSG-nblk (non-blocking), n = 3–5, 3.2 ± 0.4 MBq/15–16 μg/200 μl) and [64Cu] Keytruda (groups = 3; NSG-ctl, NSG-blk (blocking), and hNSG-nblk; n = 4, 7.4 ± 0.4 MBq /20-25 μg/200 μl) were administered in mice. PET-CT scans were performed over 1–144 h ([89Zr] Keytruda) and 1–48 h ([64Cu] Keytruda) on mice. hNSG mice exhibited a high tracer uptake in the spleen, lymphoid organs and tumors. At 24 h, human TILs homing into melanoma of hNSG-nblk mice exhibited high signal (mean %ID/g ± SD) of 3.8 ± 0.4 ([89Zr] Keytruda), and 6.4 ± 0.7 ([64Cu] Keytruda), which was 1.5- and 3-fold higher uptake compared to NSG-ctl mice (p = 0.01), respectively. Biodistribution measurements of hNSG-nblk mice performed at 144 h ([89Zr] Keytruda) and 48 h ([64Cu] Keytruda) p.i. revealed tumor to muscle ratios as high as 45- and 12-fold, respectively.

Conclusions

Our immunoPET study clearly demonstrates specific imaging of human PD-1-expressing TILs within the tumor and lymphoid tissues. This suggests these anti-human-PD-1 tracers could be clinically translatable to monitor cancer treatment response to IC blockade therapy.

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Acknowledgements

We would like to thank The Canary Foundation, The Ben and Catherine Ivy Foundation, and the National Cancer Institute for their support and for helping to fund this research. We acknowledge the supports of Drs. Mark Stolowitz, Timothy Doyle, Frezghi Habte, and Lingyun Xu; Sindhuja Ramakrishnan; and Michelle Tran for the experiments performed. MicroPET/CT imaging and gamma counter measurements were performed in the SCi3 Stanford Small Animal Imaging Service Center.

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Authors and Affiliations

  1. Department of Radiology, School of Medicine, Stanford University, Stanford, CA, USA

    Arutselvan Natarajan, Aaron T. Mayer, Robert E. Reeves & Sanjiv Sam Gambhir

  2. Department of Bioengineering, Stanford University, Stanford, CA, USA

    Aaron T. Mayer & Sanjiv Sam Gambhir

  3. Department of Comparative Medicine, Stanford University, Stanford, CA, USA

    Claude M. Nagamine

  4. Department of Materials Science and Engineering, Stanford University, Stanford, CA, USA

    Sanjiv Sam Gambhir

  5. Molecular Imaging Program at Stanford, Department of Radiology, Stanford University, James H. Clark Center, 318 Campus Drive, E153, Stanford, CA, 94305, USA

    Sanjiv Sam Gambhir

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  1. Arutselvan Natarajan
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Correspondence to Sanjiv Sam Gambhir.

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Natarajan, A., Mayer, A.T., Reeves, R.E. et al. Development of Novel ImmunoPET Tracers to Image Human PD-1 Checkpoint Expression on Tumor-Infiltrating Lymphocytes in a Humanized Mouse Model. Mol Imaging Biol 19, 903–914 (2017). https://doi.org/10.1007/s11307-017-1060-3

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