Skip to main content

Advertisement

SpringerLink
Log in

Comparative Analysis of Human Nucleoside Kinase-Based Reporter Systems for PET Imaging

  • Research Article
  • Published:
Molecular Imaging and Biology Aims and scope Submit manuscript

Abstract

Purpose

Radionuclide-based reporter gene imaging has the sensitivity to monitor gene- and cell-based therapies in human subjects. Potential immunogenicity of current viral transgenes warrants development of human-based reporter systems. We compared human nucleoside kinase reporters to a panel of nucleoside analogs of FEAU, FMAU, and FIAU, including the first in vivo assessment of l-[18F]FEAU.

Procedures

Human isogenic U87 cell lines were transduced to express different human reporter genes including dCK-R104M/D133A (dCKDM), dCK-R104Q/D133N (dCKep16A), dCK-A100V/R104M/D133A (dCK3M), and TK2-N93D/L109F (TK2DM), and wild-type dCK (dCK) and herpes simplex virus type-1 (HSVTK) reporter gene as references. In vitro cell uptake assays were performed with [18F]FEAU, l-[18F]FEAU, [14C]FMAU, l-[18F]FMAU, and [124I]FIAU. Micro-positron emission tomography/X-ray computed tomography imaging of xenograft-bearing nu/nu mice was conducted with [18F]FEAU, l-[18F]FEAU, l-[18F]FMAU, and [124I]FIAU on consecutive days. A cell viability assay was also performed to assess sensitivities to gemcitabine and bromovinyldeoxyuridine (BVdU).

Results

In vitro, dCKep16A and dCKDM with [18F]FEAU exhibited the highest sensitivity and selectivity of the human reporters, second only to HSVTK/[18F]FEAU. l-[18F]FEAU biodistribution in mice was on par with [18F]FEAU and l-[18F]FMAU. l-[18F]FMAU uptake in isogenic xenografts was highest for all human reporter genes. However, [18F]FEAU was the most selective of the short half-life reporter probes due to its minimal recognition by human dCK and relative sensitivity, whereas [124I]FIAU permitted imaging at a later time point, improving signal-to-background ratio. Of the human reporter genes, dCKep16A consistently outperformed the other tested reporters. Reporter genes of interest increased potency to the nucleoside analog prodrugs gemcitabine and BVdU.

Conclusions

We demonstrate that human nucleoside kinase reporter systems vary significantly in their sensitivity and selectivity for in vivo imaging. The sufficiently high signal-to-background ratios and enhanced suicide gene potential support clinical translation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Brader P, Serganova I, Blasberg RG (2013) Noninvasive molecular imaging using reporter genes. J Nucl Med: Off Publ Soc Nucl Med 54:167–172

    Article  CAS  Google Scholar 

  2. Koehne G, Doubrovin M, Doubrovina E et al (2003) Serial in vivo imaging of the targeted migration of human HSV-TK-transduced antigen-specific lymphocytes. Nat Biotechnol 21:405–413

    Article  CAS  PubMed  Google Scholar 

  3. Likar Y, Dobrenkov K, Olszewska M et al (2008) A new acycloguanosine-specific supermutant of herpes simplex virus type 1 thymidine kinase suitable for PET imaging and suicide gene therapy for potential use in patients treated with pyrimidine-based cytotoxic drugs. J Nucl Med : Off Publ Soc Nucl Med 49:713–720

    Article  CAS  Google Scholar 

  4. 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 U S A 97:2785–2790

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Yaghoubi SS, Gambhir SS (2006) PET imaging of herpes simplex virus type 1 thymidine kinase (HSV1-tk) or mutant HSV1-sr39tk reporter gene expression in mice and humans using [18F]FHBG. Nat Protoc 1:3069–3075

    Article  CAS  PubMed  Google Scholar 

  6. Miyagawa T, Gogiberidze G, Serganova I et al (2008) Imaging of HSV-tk Reporter gene expression: comparison between [18F]FEAU, [18F]FFEAU, and other imaging probes. J Nucl Med: Off Publ Soc Nucl Med 49:637–648

    Article  CAS  Google Scholar 

  7. Su H, Chang DS, Gambhir SS, Braun J (2006) Monitoring the antitumor response of naive and memory CD8 T cells in RAG1−/− mice by positron-emission tomography. J Immunol 176:4459–4467

    Article  CAS  PubMed  Google Scholar 

  8. Yaghoubi SS, Jensen MC, Satyamurthy N et al (2009) Noninvasive detection of therapeutic cytolytic T cells with 18F-FHBG PET in a patient with glioma. Nat Clin Pract Oncol 6:53–58

    Article  CAS  PubMed  Google Scholar 

  9. Slovin SF, Wang X, Hullings M et al (2013) Chimeric antigen receptor (CAR+) modified T cells targeting prostate-specific membrane antigen (PSMA) in patients (pts) with castrate metastatic prostate cancer (CMPC). ASCO Meet Abstr 31:72

    Google Scholar 

  10. Sabini E, Ort S, Monnerjahn C, Konrad M, Lavie A (2003) Structure of human dCK suggests strategies to improve anticancer and antiviral therapy. Nat Struct Biol 10:513–519

    Article  CAS  PubMed  Google Scholar 

  11. Iyidogan P, Lutz S (2008) Systematic exploration of active site mutations on human deoxycytidine kinase substrate specificity. Biochemistry 47:4711–4720

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Hazra S, Ort S, Konrad M, Lavie A (2010) Structural and kinetic characterization of human deoxycytidine kinase variants able to phosphorylate 5-substituted deoxycytidine and thymidine analogues. Biochemistry 49:6784–6790

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Ponomarev V, Doubrovin M, Shavrin A et al (2007) A human-derived reporter gene for noninvasive imaging in humans: mitochondrial thymidine kinase type 2. J Nucl Med: Off Publ Soc Nucl Med 48:819–826

    Article  CAS  Google Scholar 

  14. Campbell DO, Yaghoubi SS, Su Y et al (2012) Structure-guided engineering of human thymidine kinase 2 as a positron emission tomography reporter gene for enhanced phosphorylation of non-natural thymidine analog reporter probe. J Biol Chem 287:446–454

    Article  CAS  PubMed  Google Scholar 

  15. Likar Y, Zurita J, Dobrenkov K et al (2010) A new pyrimidine-specific reporter gene: a mutated human deoxycytidine kinase suitable for PET during treatment with acycloguanosine-based cytotoxic drugs. J Nucl Med: Off Publ Soc Nucl Med 51:1395–1403

    Article  CAS  Google Scholar 

  16. McCracken MN, Gschweng EH, Nair-Gill E et al (2013) Long-term in vivo monitoring of mouse and human hematopoietic stem cell engraftment with a human positron emission tomography reporter gene. Proc Natl Acad Sci U S A 110:1857–1862

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Chung J-K, Kang J, Kang K (2010) Reporter Gene Imaging with PET/SPECT. In: Gambhir SS (ed) Molecular Imaging with Reporter Genes. Cambridge University Press, Cambridge, pp 70–87

    Chapter  Google Scholar 

  18. Zhang H, Cantorias MV, Pillarsetty N, Burnazi EM, Cai S, Lewis JS (2012) An improved strategy for the synthesis of [(1)(8)F]-labeled arabinofuranosyl nucleosides. Nucl Med Biol 39:1182–1188

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Loening AM, Gambhir SS (2003) AMIDE: a free software tool for multimodality medical image analysis. Mol Imaging 2:131–137

    Article  PubMed  Google Scholar 

  20. Gil JS, Machado HB, Campbell DO et al (2013) Application of a rapid, simple, and accurate adenovirus-based method to compare PET reporter gene/PET reporter probe systems. Mol Imaging Biol: MIB: Off Publ Acad Mol Imaging 15:273–281

    Article  Google Scholar 

  21. Tseng JR, Dandekar M, Subbarayan M et al (2005) Reproducibility of 3'-deoxy-3'-(18)F-fluorothymidine microPET studies in tumor xenografts in mice. J Nucl Med: Off Publ Soc Nucl Med 46:1851–1857

    CAS  Google Scholar 

  22. Lee DJ, Prensky W, Krause G, Hughes WL (1976) Blood thymidine level and iododeoxyuridine incorporation and reutilization in DNA in mice given long-acting thymidine pellets. Cancer Res 36:4577–4583

    CAS  PubMed  Google Scholar 

  23. Radu CG, Shu CJ, Nair-Gill E et al (2008) Molecular imaging of lymphoid organs and immune activation by positron emission tomography with a new [18F]-labeled 2'-deoxycytidine analog. Nat Med 14:783–788

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Nishii R, Volgin AY, Mawlawi O et al (2008) Evaluation of 2'-deoxy-2'-[18F]fluoro-5-methyl-1-beta-L: −arabinofuranosyluracil ([18F]-L: −FMAU) as a PET imaging agent for cellular proliferation: comparison with [18F]-D: −FMAU and [18F]FLT. Eur J Nucl Med Mol Imaging 35:990–998

    Article  PubMed  Google Scholar 

  25. Tehrani OS, Douglas KA, Lawhorn-Crews JM, Shields AF (2008) Tracking cellular stress with labeled FMAU reflects changes in mitochondrial TK2. Eur J Nucl Med Mol Imaging 35:1480–1488

    Article  CAS  PubMed  Google Scholar 

  26. De Clercq E (2004) Discovery and development of BVDU (brivudin) as a therapeutic for the treatment of herpes zoster. Biochem Pharmacol 68:2301–2315

    Article  PubMed  Google Scholar 

  27. Ponomarev V, Doubrovin M, Lyddane C et al (2001) Imaging TCR-dependent NFAT-mediated T-cell activation with positron emission tomography in vivo. Neoplasia 3:480–488

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

We thank Dr. Caius G. Radu at the Ahmanson Translational Imaging Division at UCLA for donating the pMSCV-hTK2 plasmid. We thank Dr. Pat Zanzonico and Valerie Longo at Memorial Sloan Kettering Cancer Center (MSKCC) and Waldemar Ladno and Dr. Olga Sergeeva at the UCLA Crump Institute for Molecular Imaging for their imaging technical assistance and expertise. We thank Eva M. Burnazi at MSKCC for her radiochemistry technical expertise. We thank Dr. Ronald Blasberg for his help in preparing this manuscript. Technical services were provided by the MSKCC Small Animal Imaging Core Facility and the UCLA Crump Institute’s Preclinical Imaging Technology Center.

Author information

Authors and Affiliations

  1. Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, David Geffen School of Medicine at UCLA, California NanoSystems Institute Rm 2151, 570 Westwood Plaza, Los Angeles, CA, 90095, USA

    Jason T. Lee, Jeffrey Collins & R. Michael van Dam

  2. Department of Radiology, Memorial Sloan Kettering Cancer Center, 1275 York Ave, New York, NY, 10065, USA

    Jason T. Lee, Hanwen Zhang, Maxim A. Moroz, Yury Likar, Larissa Shenker, Nikita Sumzin, Jose Lobo, Juan Zurita & Vladimir Ponomarev

  3. Sloan Kettering Institute, Molecular Pharmacology and Chemistry Program, Memorial Sloan Kettering Cancer Center, New York, NY, 10065, USA

    Vladimir Ponomarev

  4. Molecular Imaging Laboratory, Department of Radiology, Memorial Sloan Kettering Cancer Center, 1275 York Ave, New York, NY, 10065, USA

    Vladimir Ponomarev

Authors
  1. Jason T. Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hanwen Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Maxim A. Moroz
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yury Likar
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Larissa Shenker
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Nikita Sumzin
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jose Lobo
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Juan Zurita
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Jeffrey Collins
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. R. Michael van Dam
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Vladimir Ponomarev
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Vladimir Ponomarev.

Ethics declarations

Funding

This work was supported by the Molecular Imaging: Training for Oncology (MITO) of the National Institutes of Health, Cancer Education and Career Development Program 5R25CA096945-09 grant, NIH P50 CA86438-11, R01 CA161138, and R01 CA163980 grants. Technical services provided by the MSKCC Small-Animal Imaging Core Facility are supported in part by NIH Small-Animal Imaging Research Program (SAIRP), NIH Shared Instrumentation Grant No 1 S10 RR020892-01, NIH Shared Instrumentation Grant No 1 S10 RR028889-01, and NIH Center Grant P30 CA08748. Technical services provided by the UCLA Crump Institute’s Preclinical Imaging Technology Center are supported in part by NIH In Vivo Cellular and Molecular Imaging Centers Grant P50 CA086306, NIH Cancer Center Support Grant P30 CA016042, and NIH SPORE Grant P50 CA092131.

Conflict of Interest

The authors declare that they have no conflict of interest.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(PDF 1280 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lee, J.T., Zhang, H., Moroz, M.A. et al. Comparative Analysis of Human Nucleoside Kinase-Based Reporter Systems for PET Imaging. Mol Imaging Biol 19, 100–108 (2017). https://doi.org/10.1007/s11307-016-0981-6

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11307-016-0981-6

Key words

Search

Navigation