Abstract
Positron emission tomography (PET) reporter genes allow noninvasive whole-body imaging of transplanted cells by detection with radiolabeled probes. We used a human deoxycytidine kinase containing three amino acid substitutions within the active site (hdCK3mut) as a reporter gene in combination with the PET probe [(18)F]-L-FMAU (1-(2-deoxy-2-(18)fluoro-β-L-arabinofuranosyl)-5-methyluracil) to monitor models of mouse and human hematopoietic stem cell (HSC) transplantation. These mutations in hdCK3mut expanded the substrate capacity allowing for reporter-specific detection with a thymidine analog probe. Measurements of long-term engrafted cells (up to 32 wk) demonstrated that hdCK3mut expression is maintained in vivo with no counter selection against reporter-labeled cells. Reporter cells retained equivalent engraftment and differentiation capacity being detected in all major hematopoietic lineages and tissues. This reporter gene and probe should be applicable to noninvasively monitor therapeutic cell transplants in multiple tissues.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Arabinofuranosyluracil / analogs & derivatives
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Arabinofuranosyluracil / chemistry
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Arabinofuranosyluracil / metabolism
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Blotting, Western
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Cell Line, Tumor
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Deoxycytidine Kinase / genetics
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Deoxycytidine Kinase / metabolism*
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Female
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Fluorine Radioisotopes / chemistry
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Hematopoietic Stem Cell Transplantation / methods*
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Hematopoietic Stem Cells / diagnostic imaging*
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Hematopoietic Stem Cells / metabolism
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Immunohistochemistry
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Interleukin Receptor Common gamma Subunit / deficiency
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Interleukin Receptor Common gamma Subunit / genetics
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Kaplan-Meier Estimate
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Luminescent Proteins / genetics
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Luminescent Proteins / metabolism
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Mice
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Mice, Inbred C57BL
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Mice, Inbred NOD
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Mice, Inbred Strains
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Mice, Knockout
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Mice, SCID
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Mutation
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Positron-Emission Tomography / methods*
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Thymus Gland / diagnostic imaging
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Thymus Gland / metabolism
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Time Factors
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Transplantation, Heterologous
Substances
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Fluorine Radioisotopes
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Il2rg protein, mouse
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Interleukin Receptor Common gamma Subunit
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Luminescent Proteins
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Arabinofuranosyluracil
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Deoxycytidine Kinase
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clevudine