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Single-Cell Imaging Using Radioluminescence Microscopy Reveals Unexpected Binding Target for [18F]HFB

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Abstract

Purpose

Cell-based therapies are showing great promise for a variety of diseases, but remain hindered by the limited information available regarding the biological fate, migration routes and differentiation patterns of infused cells in trials. Previous studies have demonstrated the feasibility of using positron emission tomography (PET) to track single cells utilising an approach known as positron emission particle tracking (PEPT). The radiolabel hexadecyl-4-[18F]fluorobenzoate ([18F]HFB) was identified as a promising candidate for PEPT, due to its efficient and long-lasting labelling capabilities. The purpose of this work was to characterise the labelling efficiency of [18F]HFB in vitro at the single-cell level prior to in vivo studies.

Procedures

The binding efficiency of [18F]HFB to MDA-MB-231 and Jurkat cells was verified in vitro using bulk gamma counting. The measurements were subsequently repeated in single cells using a new method known as radioluminescence microscopy (RLM) and binding of the radiolabel to the single cells was correlated with various fluorescent dyes.

Results

Similar to previous reports, bulk cell labelling was significantly higher with [18F]HFB (18.75 ± 2.47 dpm/cell, n = 6) than 2-deoxy-2-[18F]fluoro-D-glucose ([18F]FDG) (7.59 ± 0.73 dpm/cell, n = 7; p ≤ 0.01). However, single-cell imaging using RLM revealed that [18F]HFB accumulation in live cells (8.35 ± 1.48 cpm/cell, n = 9) was not significantly higher than background levels (4.83 ± 0.52 cpm/cell, n = 12; p > 0.05) and was 1.7-fold lower than [18F]FDG uptake in the same cell line (14.09 ± 1.90 cpm/cell, n = 13; p < 0.01). Instead, [18F]HFB was found to bind significantly to fragmented membranes associated with dead cell nuclei, suggesting an alternative binding target for [18F]HFB.

Conclusion

This study demonstrates that bulk analysis alone does not always accurately portray the labelling efficiency, therefore highlighting the need for more routine screening of radiolabels using RLM to identify heterogeneity at the single-cell level.

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Funding

This work was funded in part by NIH grants to Dr. Pratx (5R21HL127900 and 5R01CA186275) and by a NIH training grant to Dr. Kim (T32CA11868).

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Authors and Affiliations

  1. Department of Radiation Oncology, Stanford University School of Medicine, Stanford, CA, USA

    Louise Kiru, Tae Jin Kim & Guillem Pratx

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

    Bin Shen & Frederick T. Chin

Authors
  1. Louise Kiru
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  2. Tae Jin Kim
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  3. Bin Shen
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  4. Frederick T. Chin
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  5. Guillem Pratx
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Correspondence to Guillem Pratx.

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Research Involving Human Participants and/or Animals

This article does not contain any studies with human participants or animals performed by any of the authors.

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Kiru, L., Kim, T.J., Shen, B. et al. Single-Cell Imaging Using Radioluminescence Microscopy Reveals Unexpected Binding Target for [18F]HFB. Mol Imaging Biol 20, 378–387 (2018). https://doi.org/10.1007/s11307-017-1144-0

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