Stratification of nucleoside analog chemotherapy using 1-(2'-deoxy-2'-18F-fluoro-β-D-arabinofuranosyl)cytosine and 1-(2'-deoxy-2'-18F-fluoro-β-L-arabinofuranosyl)-5-methylcytosine PET.
The ability to measure tumor determinants of response to nucleoside analog (NA) chemotherapy agents such as gemcitabine and related compounds could significantly affect the management of several types of cancer. Previously we showed that the accumulation in tumors of the new PET tracer 1-(2'-deoxy-2'-(18)F-fluoro-β-d-arabinofuranosyl)cytosine ((18)F-FAC) is predictive of responses to gemcitabine. (18)F-FAC retention in cells requires deoxycytidine kinase (dCK), a rate-limiting enzyme in the deoxyribonucleoside salvage metabolism and in gemcitabine conversion from an inactive prodrug to a cytotoxic compound. The objectives of the current study were to determine whether (18)F-FAC tumor uptake is also influenced by cytidine deaminase (CDA), a determinant of resistance to gemcitabine; to develop a new PET assay using (18)F-FAC and the related probe 1-(2'-deoxy-2'-(18)F-fluoro-β-l-arabinofuranosyl)-5-methylcytosine (l-(18)F-FMAC) to profile tumor lesions for both dCK and CDA enzymatic activities; and to determine whether this PET assay can identify the most effective NA chemotherapy against tumors with differential expression of dCK and CDA.


METHODS
Isogenic murine leukemic cell lines with defined dCK and CDA activities were generated by retroviral transduction. A cell viability assay was used to determine the sensitivity of the isogenic cell lines to the dCK-dependent NA prodrugs gemcitabine and clofarabine. In vitro enzymatic and cell-based tracer uptake assays and in vivo PET with (18)F-FAC and l-(18)F-FMAC were used to predict tumor responses to gemcitabine and clofarabine.


RESULTS
dCK and CDA activities measured by kinase and tracer uptake assays correlated with the sensitivity of isogenic cell lines to gemcitabine and clofarabine. Coexpression of CDA decreased the sensitivity of dCK-positive cells to gemcitabine treatment in vitro by 15-fold but did not affect responses to clofarabine. Coexpression of CDA decreased (18)F-FAC but not l-(18)F-FMAC, phosphorylation, and uptake by dCK-positive cells. (18)F-FAC and l-(18)F-FMAC PET estimates of the enzymatic activities of dCK and CDA in tumor implants in mice were predictive of responses to gemcitabine and clofarabine treatment in vivo.


CONCLUSIONS
These findings support the utility of PET-based phenotyping of tumor nucleoside metabolism for guiding the selection of NA prodrugs.
Ahmanson Translational Imaging Division, Department of Molecular and Medical Pharmacology, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.
Jason T
JT
Lee
Dean O
DO
Campbell
Nagichettiar
N
Satyamurthy
Johannes
J
Czernin
Caius G
CG
Radu
D000227:Adenine Nucleotides / Q000494:pharmacology / Q000627:therapeutic use*; D000818:Animals; D001087:Arabinonucleosides / Q000494:pharmacology / Q000627:therapeutic use*; D045744:Cell Line, Tumor; D000077866:Clofarabine; D003561:Cytarabine / Q000031:analogs & derivatives*; D003564:Cytidine Deaminase / Q000378:metabolism; D003596:Cytosine; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use; D003842:Deoxycytidine Kinase / Q000378:metabolism; D005462:Fluorine Radioisotopes; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D007945:Leukemia, Lymphoid / Q000000981:diagnostic imaging / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism; D051379:Mice; D049268:Positron-Emission Tomography / Q000379:methods*; D016896:Treatment Outcome; D000093542:Gemcitabine
C571030:1-(2'-deoxy-2'-fluoro-arabinofuranosyl)cytosine; D000227:Adenine Nucleotides; D001087:Arabinonucleosides; D005462:Fluorine Radioisotopes; D003561:Cytarabine; D003841:Deoxycytidine; D000077866:Clofarabine; C034934:2'-fluoro-5-methylarabino-furanosylcytosine; D003596:Cytosine; D003842:Deoxycytidine Kinase; D003564:Cytidine Deaminase; D000093542:Gemcitabine