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Comparison of Near-Infrared Imaging Agents Targeting the PTPmu Tumor Biomarker

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Abstract

Purpose

Maximal, safe resection of solid tumors is considered a critical first step in successful cancer treatment. The advent of fluorescence image-guided surgery (FIGS) using non-specific agents has improved patient outcomes, particularly in the case of glioblastoma. Molecularly targeted agents that recognize specific tumor biomarkers have the potential to augment these gains. Identification of the optimal combination of targeting moiety and fluorophore is needed prior to initiating clinical trials.

Procedures

A 20-amino acid peptide (SBK2) recognizing the receptor protein-tyrosine phosphatase mu (PTPmu)–derived tumor-specific biomarker, with or without a linker, was conjugated to three different near-infrared fluorophores: indocyanine green (ICG), IRDye® 800CW, and Tide Fluor™ 8WS. The in vivo specificity, time course, and biodistribution were evaluated for each using mice with heterotopic human glioma tumors that express the PTPmu biomarker to identify component combinations with optimal properties for FIGS.

Results

SBK2 conjugated to ICG demonstrated excellent specificity for gliomas in heterotopic tumors. SBK2-ICG showed significantly higher in vivo tumor labeling compared to the Scram-ICG control from 10 min to 24 h, p < 0.01 at all timepoints, following injection, as well as a significantly higher ex vivo tumor signal at 24 h, p < 0.001. Inserting a six-amino acid linker between the targeting peptide and ICG increased the clearance rate and resulted in significantly higher in vivo tumor signal relative to its linker-containing Scrambled control from 10 min to 8 h, p < 0.05 at all timepoints, after dosing. Agents made with the more hydrophilic IRDye® 800CW and Tide Fluor™ 8WS showed no specific tumor labeling relative to the controls. The IRDye 800CW-conjugated agents cleared within 1 h, while the non-specific fluorescent tumor signal generated by the Tide Fluor 8WS-conjugated agents persists beyond 24 h.

Conclusions

The SBK2 PTPmu-targeting peptide conjugated to ICG specifically labels heterotopic human gliomas grown in mice between 10 min and 24 h following injection. Similar molecules constructed with more hydrophilic dyes demonstrated no specificity. These studies present a promising candidate for use in FIGS of PTPmu biomarker–expressing tumors.

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Data Availability

The data that support the findings of this study are available from the corresponding author (SBK) upon reasonable request.

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Acknowledgements

We thank Jennifer Major and Mamuni Swain for expert technical assistance, and Zoey Lockwood for assistance with the Varian spectrophotometers. An NIH shared instrument grant, 1S10RR031537-01, purchased the Orbitrap Elite LC-MS that was utilized for these agents in the Lerner Research Institute Proteomics Core at the Cleveland Clinic. SBK and AES were funded by a National Institutes of Health grant (R01CA217956). SBK was funded by the Tabitha Yee-May Lou Endowment Fund for Brain Cancer Research. Additional support was obtained from the National Institutes of Health sponsored Case Comprehensive Cancer Center, their Cancer Imaging Program and its cores (P30 CA043703).

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Authors and Affiliations

  1. Department of Molecular Biology and Microbiology, Case Western Reserve University School of Medicine, 10900 Euclid Ave., Cleveland, OH, 44106, USA

    Mette L. Johansen, Jason Vincent, Marissa Rose & Susann M. Brady-Kalnay

  2. Department of Neurological Surgery, Case Western Reserve University and University Hospitals, Cleveland, OH, 44106, USA

    Andrew E. Sloan

  3. The Case Comprehensive Cancer Center, Case Western Reserve University, 10900 Euclid Ave., Cleveland, OH, 44106, USA

    Andrew E. Sloan & Susann M. Brady-Kalnay

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Correspondence to Susann M. Brady-Kalnay.

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Conflict of Interest

Dr. Brady-Kalnay has licensed these agents to the biotechnology company NeoIndicate, where she serves as the Chief Scientific Officer. Mette Johansen is an inventor on the relevant patents.

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Johansen, M.L., Vincent, J., Rose, M. et al. Comparison of Near-Infrared Imaging Agents Targeting the PTPmu Tumor Biomarker. Mol Imaging Biol 25, 744–757 (2023). https://doi.org/10.1007/s11307-023-01799-5

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