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124I-PET Assessment of Human Sodium Iodide Symporter Reporter Gene Activity for Highly Sensitive In Vivo Monitoring of Teratoma Formation in Mice

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Abstract

Purpose

Pluripotent stem cell (PSC)-based therapies possess great potential to restore the function of irreversibly damaged organs. PSCs can be differentiated in vitro into any cell type. However, pluripotent potential bears the risk of teratoma formation. In vivo monitoring of teratoma formation is indispensable, as 100 % purity of the cell preparation cannot be achieved. We aimed at establishing the human sodium iodide symporter (hNIS) as reporter gene for PET monitoring of teratoma formation.

Procedures

Murine PSC stably expressing hNIS were injected into the hind limbs of SCID mice to induce teratoma formation. Positron emission tomography (PET) scans were acquired weekly between days 14 and 42 after transplantation. Two teratomas were excised at each time point for histology and size measurement. Tracer uptake was correlated with teratoma weight. Specificity of tumoural iodine uptake was assessed by blocking hNIS in vivo with perchlorate.

Results

Neither hNIS expression nor I-124 exposure adversely impacted viability or differentiation potential of PSCs. Iodine uptake was highly specific in teratomas, as in vivo blocking of hNIS with perchlorate led to uptake rates comparable to tracer uptake in non-transgene tumours. Tumour mass and tracer uptake showed a positive correlation.

Conclusions

This is the first study to generate stably hNIS-expressing murine PSCs. Since the differentiation potential was preserved, hNIS-expressing cells are suitable for PSC-based forward programming approaches. Teratoma formation from undifferentiated cells can be monitored in vivo by PET with high specificity on a quantitative level. Due to its anticipated lack of immunogenicity in humans, hNIS is a promising reporter gene for clinical translation.

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Abbreviations

%ID:

Percentage of the injected dose

I-124:

Iodine-124

I-125:

Iodine-125

BMSC:

Bone marrow stem cells

DMEM:

Dulbecco’s modified eagle medium

EB:

Embryoid body

ESC:

Embryonic stem cells

[18F]FDG:

2-Deoxy-2-[18F]fluoro-d-glucose

hNIS:

Human sodium iodide symporter

iPS:

Induced pluripotent stem cells

LAD:

Left anterior descending artery

MAP:

Maximum a posteriori

MI:

Myocardial infarction

OSEM:

Ordered subset expectation maximization

PBS:

Phosphate buffered saline

PET:

Positron emission tomography

PFA:

Paraformaldehyde

PSC:

Pluripotent stem cells

SCID:

Severe combined immunodeficiency

VOI:

Volume of interest

[18F]FHBG:

9-(4-[18F]fluoro-3-[hydroxymethyl]butyl)guanine

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Acknowledgments

We thank Christiane Groß, Barbara Markieton and Judith Arcifa for the expert technical assistance. This work was supported by the FöFoLe Program of the LMU Munich [C.L. and W.M.F.], the BMBF [01GN0960 to R.D. and W.M.F.] and the Deutsche Forschungsgemeinschaft [DA 1296/2-1 to R.D. and FR 705/14-2 to W.M.F.]. This work has been funded by the Federal Ministry of Education and Research Germany (FKZ 0312138A and FKZ 316159) and the State Mecklenburg-Western Pomerania with EU Structural Funds (ESF/IV-WM-B34-0030/10 and ESF/IV-BM-B35-0010/12), by the DFG (DA 1296-1) and the German Heart Foundation (F/01/12). A substantial part of this work originated from the doctoral thesis of Georgios Kaissis.

Conflict of Interest

The authors declare that they have no conflict of interest.

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    Authors and Affiliations

    1. Department of Nuclear Medicine, University of Munich, Munich, Germany

      Sebastian Lehner, Georgios Kaissis, Andrei Todica, Mathias Johannes Zacherl, Guido Boening & Peter Bartenstein

    2. Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), University of Rostock, Rostock, Germany

      Cajetan Lang & Robert David

    3. University Clinic of Internal Medicine III, Cardiology, Innsbruck Medical University, Innsbruck, Austria

      Wolfgang-Michael Franz

    4. Munich Heart Alliance, University of Munich, Munich, Germany

      Wolfgang-Michael Franz

    5. Department of Cardiac Surgery, Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), University of Rostock, Rostock, Germany

      Gustav Steinhoff

    6. Department of Internal Medicine II—Campus Grosshadern, University of Munich, Munich, Germany

      Christine Spitzweg

    7. Department of Biomedical Imaging and Image-Guided Therapy, Division of Nuclear Medicine, Radiopharmacy and Experimental Nuclear Medicine, Medical University of Vienna, Vienna, Austria

      Marcus Hacker

    8. Department of Nuclear Medicine, University of Rostock, Rostock, Germany

      Bernd Joachim Krause

    9. Institute of Veterinary Pathology, University of Munich, Munich, Germany

      Nadja Herbach

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    Correspondence to Robert David.

    Additional information

    Robert David and Marcus Hacker share senior authorships.

    Sebastian Lehner and Cajetan Lang contributed equally to this work.

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    EB’s from transgenic ESC’s retained their cardiogenic potential in vitro, as evident from spontaneously contracting foci that occurred on day 12 of differentiation (A,B). (M4V 5188 kb)

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    Lehner, S., Lang, C., Kaissis, G. et al. 124I-PET Assessment of Human Sodium Iodide Symporter Reporter Gene Activity for Highly Sensitive In Vivo Monitoring of Teratoma Formation in Mice. Mol Imaging Biol 17, 874–883 (2015). https://doi.org/10.1007/s11307-015-0857-1

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