Cystathionine-beta-synthase cDNA transfection alters the sensitivity and metabolism of 1-beta-D-arabinofuranosylcytosine in CCRF-CEM leukemia cells in vitro and in vivo: a model of leukemia in Down syndrome.
The significantly higher event-free survival rates of Down syndrome (DS) children with acute myeloid leukemia compared with non-DS children is linked to increased sensitivity of DS myeloblasts to 1-beta-D-arabinofuranosylcytosine (ara-C) and the enhanced metabolism of ara-C to ara-C triphosphate (J. W. Taub et al., Blood, 87: 3395-3403, 1996). The cystathionine-beta-synthase (CBS) gene (localized to chromosome 21q22.3) may have downstream effects on reduced folate and S-adenosylmethionine pathways; ara-C metabolism and folate pools are linked by the known synergistic effect of sequential methotrexate and ara-C therapy. We have shown that relative CBS transcripts were significantly higher in DS compared with non-DS myeloblasts, and CBS transcript levels correlated with in vitro ara-C sensitivity (J. W. Taub et al., Blood, 94: 1393-1400, 1999). A leukemia cell line model to study the relationship of the CBS gene and ara-C metabolism/sensitivity was developed by transfecting CBS-null CCRF-CEM cells with the CBS cDNA. CBS-transfected cells were a median 15-fold more sensitive in vitro to ara-C compared with wild-type cells and generated 8.5-fold higher [3H]ara-C triphosphate levels after in vitro incubation with [3H]ara-C. Severe combined immunodeficient mice implanted with CBS-transfected CEM cells demonstrated greater responsiveness to therapy, reflected in significantly prolonged survivals after ara-C administration compared with mice implanted with wild-type cells and treated with the same dosage schedule. The transfected cells also demonstrated increased in vitro and in vivo sensitivity to gemcitabine. Deoxycytidine kinase (dCK) activity was approximately 22-fold higher in transfected CEM cells compared with wild-type cells. However, levels of dCK transcripts on Northern blots and protein levels on Western blots were nearly identical between CBS-transfected and wild-type cells. Collectively, these results suggest a posttranscriptional regulation of dCK in CBS-overexpressing cells that contributes to increased ara-C phosphorylation and drug activity. Further elucidating the mechanisms of increased sensitivity of DS cells to ara-C related to the CBS gene may lead to the application of these novel approaches to acute myeloid leukemia therapy for non-DS patients.
Taub
J W
JW
Division of Pediatric Hematology/Oncology, Children's Hospital of Michigan, Detroit 48201, USA.
Huang
X
X
Ge
Y
Y
Dutcher
J A
JA
Stout
M L
ML
Mohammad
R M
RM
Ravindranath
Y
Y
Matherly
L H
LH
D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000378:metabolism / Q000494:pharmacology*; D002891:Chromosomes, Human, Pair 21 / Q000235:genetics; D003541:Cystathionine beta-Synthase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D003561:Cytarabine / Q000378:metabolism / Q000494:pharmacology*; D018076:DNA, Complementary / Q000235:genetics*; D003842:Deoxycytidine Kinase / Q000378:metabolism; D004314:Down Syndrome / Q000150:complications* / Q000201:enzymology / Q000235:genetics; D005260:Female; D015870:Gene Expression; D006801:Humans; D007942:Leukemia, Experimental / Q000188:drug therapy / Q000201:enzymology* / Q000235:genetics; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D016513:Mice, SCID; D014162:Transfection; D014407:Tumor Cells, Cultured; D023041:Xenograft Model Antitumor Assays
D000964:Antimetabolites, Antineoplastic; D018076:DNA, Complementary; D003561:Cytarabine; D003842:Deoxycytidine Kinase; D003541:Cystathionine beta-Synthase