Molecular Imaging and Biology

, Volume 19, Issue 6, pp 903–914 | Cite as

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

  • Arutselvan Natarajan
  • Aaron T. Mayer
  • Robert E. Reeves
  • Claude M. Nagamine
  • Sanjiv Sam Gambhir
Research Article

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.

Key words

ImmunoPET Tumor-infiltrating lymphocytes PD-1 Keytruda 64-Cu 89-Zr 

Notes

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.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

11307_2017_1060_MOESM1_ESM.docx (3.1 mb)
ESM 1(PDF 3.07 mb)

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Copyright information

© World Molecular Imaging Society 2017

Authors and Affiliations

  • Arutselvan Natarajan
    • 1
  • Aaron T. Mayer
    • 1
    • 2
  • Robert E. Reeves
    • 1
  • Claude M. Nagamine
    • 3
  • Sanjiv Sam Gambhir
    • 1
    • 2
    • 4
    • 5
  1. 1.Department of Radiology, School of MedicineStanford UniversityStanfordUSA
  2. 2.Department of BioengineeringStanford UniversityStanfordUSA
  3. 3.Department of Comparative MedicineStanford UniversityStanfordUSA
  4. 4.Department of Materials Science and EngineeringStanford UniversityStanfordUSA
  5. 5.Molecular Imaging Program at Stanford, Department of RadiologyStanford UniversityStanfordUSA

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