Sebastian Lehner1, Cajetan Lang2, Georgios Kaissis1, Andrei Todica1, Mathias Johannes Zacherl1, Guido Boening1, Christine Spitzweg3, Nadja Herbach4, Wolfgang-Michael Franz5,6, Bernd Joachim Krause7, Gustav Steinhoff8, Peter Bartenstein1, Marcus Hacker9, Robert David10. 1. Department of Nuclear Medicine, University of Munich, Munich, Germany. 2. Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), University of Rostock, Rostock, Germany. 3. Department of Internal Medicine II-Campus Grosshadern, University of Munich, Munich, Germany. 4. Institute of Veterinary Pathology, University of Munich, Munich, Germany. 5. University Clinic of Internal Medicine III, Cardiology, Innsbruck Medical University, Innsbruck, Austria. 6. Munich Heart Alliance, University of Munich, Munich, Germany. 7. Department of Nuclear Medicine, University of Rostock, Rostock, Germany. 8. Department of Cardiac Surgery, Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), University of Rostock, Rostock, Germany. 9. Department of Biomedical Imaging and Image-Guided Therapy, Division of Nuclear Medicine, Radiopharmacy and Experimental Nuclear Medicine, Medical University of Vienna, Vienna, Austria. 10. Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), University of Rostock, Rostock, Germany. robert.david@med.uni-rostock.de.
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.
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 humansodium 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 SCIDmice 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 tumouraliodine 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 murinePSCs. 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.
Entities:
Keywords:
In vivo monitoring; Reporter gene; Small animal PET; Sodium iodide symporter; Stem cell therapy; Teratoma formation
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