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.
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.
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.
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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Slavin S, Nagler A, Naparstek E et al (1998) Nonmyeloablative stem cell transplantation and cell therapy as an alternative to conventional bone marrow transplantation with lethal cytoreduction for the treatment of malignant and nonmalignant hematologic diseases. Blood 91:756–763
Koc NO, Gerson LS, Cooper WB et al (2016) Rapid hematopoietic recovery after coinfusion of autologous-blood stem cells and culture-expanded marrow mesenchymal stem cells in advanced breast. J Clin Oncologia 18:307–316
Maus MV, Grupp SA, Porter DL, June CH (2014) Antibody modified T cells: CARs take the front seat for hematologic malignancies. Blood 123:2625–2635
Robbins PF, Lu Y-C, El-Gamil M et al (2013) Mining exomic sequencing data to identify mutated antigens recognized by adoptively transferred tumor-reactive T cells. Nat Med 19:747–752
Hsu FJ, Benike C, Fagnoni F et al (1996) Vaccination of patients with B-cell lymphoma using autologous antigen-pulsed dendritic cells. Nat Med 2:2338–2344
Nestle FO, Alijagic S, Gilliet M et al (1998) Vaccination of melanoma patients with peptide- or tumor lysate-pulsed dendritic cells. Nat Med 4:201–206
Lo Celso C, Fleming HE, Wu JW et al (2009) Live-animal tracking of individual haematopoietic stem/progenitor cells in their niche. Nature 457:92–96
Doherty WP, Bushberg TJ, Lipton JM et al (1978) The use of indium-111-labeled leukocytes for abscess detection. Clin Nucl Med 3:108–110
Sharpe M, Mount N (2015) Genetically modified T cells in cancer therapy : opportunities and challenges. Dis Model Mech 8:337–350
Griessinger CM, Kehlbach R, Bukala D et al (2014) In vivo tracking of Th1 cells by PET reveals quantitative and temporal distribution and specific homing in lymphatic tissue. J Nucl Med 2:301–307
Yaghoubi SS, Jensen MC, Satyamurthy N et al (2012) Noninvasive detection of therapeutic cytolytic T cells with PET in a glioma patient. Nat Rev Clin Oncologia 6:53–58
Rodriguez-porcel M, Kronenberg MW, Henry TD et al (2012) Cell tracking and the development of cell-based therapies: a view from the Cadiovascular Cell Therapty Research Network. JACC Cardiovasc Imaging 5:559–565
Kircher MF, Gambhir SS, Grimm J (2011) Noninvasive cell-tracking methods. Nat Rev Clin Oncol 8:677–688
Adonai NN, Nguyen KN, Walsh J et al (2002) Ex vivo cell labeling with 64Cu-pyruvaldehyde-bis(N4-methylthiosemicarbazone) for imaging cell trafficking in mice with positron-emission tomography. Proc Natl Acad Sci 99:3030–3035
Hofmann M, Wollert KC, Meyer GP et al (2005) Monitoring of bone marrow cell homing into the infarcted human myocardium. Circulation 111:2198–2202
Gambhir SS, Bauer E, Black ME et al (2000) A mutant herpes simplex virus type 1 thymidine kinase reporter gene shows improved sensitivity for imaging reporter gene expression with positron emission tomography. Proc Natl Acad Sci 97:2785–2790
Kang JH, Lee DS, Paeng JC et al (2005) Development of a sodium/iodide symporter (NIS)-transgenic mouse for imaging of cardiomyocyte-specific reporter gene expression. J Nucl Med 46:479–483
Moroz MA, Serganova I, Zanzonico P et al (2007) Imaging hNET reporter gene expression with 124I-MIBG. J Nucl Med 48:827–836
Ma B, Hankenson KD, Dennis JE et al (2005) A simple method for stem cell labeling with fluorine 18. Nucl Med Biol 32:701–705
Lee KS, Kim TJ, Pratx G (2015) Single-cell tracking with PET using a novel trajectory reconstruction algorithm. IEEE Trans Med Imaging 34:994–1003
Langford ST, Wiggins CS, Santos R et al (2017) Three-dimensional spatiotemporal tracking of fluorine-18 radiolabeled yeast cells via positron emission particle tracking. PLoS One 12:7
Wiggins C, Santos R, Ruggles A (2016) A novel clustering approach to positron emission particle tracking. Nucl Instrum Methods Phys Res Sect A 811:18–24
Parker DJ, Broadbent CJ, Fowles P et al (1993) Positron emission particle tracking—a technique for studying flow within engineering equipment. Nucl Inst Methods Phys Res A 326:592–607
Bemrose CR, Fowles P, Hawkesworth MR, O’Dwyer MA (1988) Application of positron emission tomography to particulate flow measurement in chemical engineering processes. Nucl Instrum Methods Phys Res A 273:874–880
Patel N, Wiggins C, Ruggles A (2017) Positron emission particle tracking in pulsatile flow. Exp Fluids 58:42
Ouyang Y, Kim TJ, Pratx G (2016) Evaluation of a BGO-based PET system for single-cell tracking performance by simulation and phantom studies. Mol Imaging 15:1–8
Zhang Y, DaSilva JN, Hadizad T et al (2012) F-18-FDG cell labeling may underestimate transplanted cell homing: more accurate, efficient, and stable cell labeling with hexadecyl-4-[F-18]fluorobenzoate for in vivo tracking of transplanted human progenitor cells by positron emission tomography. Cell Transplant 21:1821–1835
Pratx G, Chen K, Sun C et al (2012) Radioluminescence microscopy: measuring the heterogeneous uptake of radiotracers in single living cells. PLoS One 7:1–9
Pratx G, Chen K, Sun C et al (2013) High-resolution radioluminescence microscopy of 18F-FDG uptake by reconstructing the β-ionization track. J Nucl Med 54:1841–1846
Kim TJ, Türkcan S, Pratx G (2017) Modular low-light microscope for imaging cellular bioluminescence and radioluminescence. Nat Protoc 12:1055–1076
Wu M, Kwon HY, Rattis F et al (2017) Imaging hematopoietic precursor division in real time. Cell Stem Cell 1:541–554
Chang HH, Hemberg M, Barahona M et al (2008) Transcriptome-wide noise controls lineage choice in mammalian progenitor cells. Nature 453:544–547
Ravin R, Hoeppner DJ, Munno DM et al (2008) Potency and fate specification in CNS stem cell populations in vitro. Cell Stem Cell 3:670–680
Naumova AV, Modo M, Moore A et al (2014) Clinical imaging in regenerative medicine. Nat Biotechnol 32:804–818
Ahmadi A, Thorn SL, Alarcon EI et al (2015) PET imaging of a collagen matrix reveals its effective injection and targeted retention in a mouse model of myocardial infarction. Biomaterials 49:18–26
Kim MH, Woo SK, Lee KC et al (2015) Longitudinal monitoring adipose-derived stem cell survival by PET imaging hexadecyl-4-124I-iodobenzoate in rat myocardial infarction model. Biochem Biophys Res Commun 456:13–19
Kim MH, Woo SK, Kim KI et al (2015) Simple methods for tracking stem cells with 64 cu-labeled DOTA-hexadecyl-benzoate. ACS Med Chem Lett 6:528–530
Natarajan A, Türkcan S, Gambhir SS, Pratx G (2015) Multiscale framework for imaging radiolabeled therapeutics. Mol Pharm 12:4554–4560
Kim TJ, Tuerkcan S, Ceballos A, Pratx G (2015) Modular platform for low-light microscopy. Biomed Opt Express 6:4585–4598
Sengupta D, Pratx G (2016) Single-cell characterization of FLT uptake with radioluminescence microscopy. J Nucl Med 2764:1136–1141
Kim MH, Lee KC, Il AG et al (2015) Evaluation of safety and efficacy of adipose-derived stem cells in rat myocardial infarction model using hexadecyl-4-[(124)I]iodobenzoate for cell tracking. Appl Radiat Isot 108:116–123
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).
Conflict of Interest
The authors declare that they have no conflict of interest.
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
- Cell tracking
- Radioluminescence microscopy (RLM)
- Single-cell imaging
- Hexadecyl-4-[18F]fluorobenzoate ([18F]HFB)