Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
Proc Natl Acad Sci U S A. 2013 Jan 29;110(5):1857-62. doi: 10.1073/pnas.1221840110. Epub 2013 Jan 14.
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.
正电子发射断层扫描（PET）报告基因允许通过放射性标记探针检测，对移植细胞进行非侵入性全身成像。我们使用在活性位点内含有三个氨基酸取代的人脱氧胞苷激酶（hdCK3mut）作为报告基因，与 PET 探针 [(18)F]-L-FMAU（1-(2-脱氧-2-(18)氟-β-L-阿拉伯呋喃糖基)-5-甲基尿嘧啶）结合，以监测小鼠和人造血干细胞（HSC）移植模型。hdCK3mut 中的这些突变扩大了底物容量，允许用胸苷类似物探针进行报告基因特异性检测。对长期植入细胞（长达 32 周）的测量表明，hdCK3mut 表达在体内得以维持，没有针对报告基因标记细胞的反选择。报告细胞在所有主要的造血谱系和组织中均保持着等效的植入和分化能力，这可以被检测到。这种报告基因和探针应该适用于非侵入性地监测多种组织中的治疗性细胞移植。
