Two Nonsynonymous Single Nucleotide Polymorphisms of Human Carbonyl Reductase 1 Demonstrate Reduced in Vitro Metabolism of Daunorubicin and Doxorubicin

Abstract

ABSTRACT:Carbonyl reductases (CBRs) are a group of metabolic enzymes belonging to the short-chain dehydrogenase family with NADPHdependent oxidoreductase activity. These enzymes are known to metabolize the anthracyclines doxorubicin (DOX) and daunorubicin (DAUN). Both DOX and DAUN are highly effective in cancer therapy; however, there is considerable interpatient variability in adverse effects seen in patients undergoing treatment with these drugs. This may be attributed to altered metabolism associated with nonsynonymous single nucleotide polymorphisms (ns-SNPs) in the genes encoding for CBRs. In this study, we examine the effect of the V88I and P131S mutations in the human CBR1 gene on the metabolism of anthracyclines to their respective major metabolites, doxorubicinol and daunorubicinol. Kinetic studies using purified, histidine-tagged, recombinant enzymes in a high-performance liquid chromatography-fluorescence assay demonstrated that the V88I mutation leads to a significantly reduced maximal rate of activity (V max ) (2090 ؎ 112 and 257 ؎ 11 nmol/min ⅐ mg of purified protein for DAUN and DOX, respectively) compared with that for the wild-type (3430 ؎ 241 and 364 ؎ 37 nmol/min ⅐ mg of purified protein for DAUN and DOX, respectively). In the case of the P131S mutation, a significant increase in substrate affinity (K m ) was observed for DAUN only (89 ؎ 13 M) compared with that for the wild-type (51 ؎ 13 M). In the presence of either anthracycline, both variants exhibited a 20 to 40% decrease in catalytic efficiency (k cat /K m ) compared with that for the wild-type enzyme. Therefore, the ns-SNPs generating both these mutations may alter bioavailability of these anthracyclines in cancer patients and should be examined in clinical studies as potential biomarkers for DAUN-and DOX-induced adverse effects.

Resveratrol Reduces the Hypoxia-Induced Resistance to Doxorubicin in Breast Cancer Cells

“…Recently, we found that resveratrol inhibits enzymatic activity of carbonyl reductase 1 (CBR1), a doxorubicin-metabolizing enzyme, by directly binding to CBR1 (14). CBR1 is an NADPH-dependent enzyme belonging to the short chain dehydrogenase family and catalyzes a large number of biologically and pharmacologically active substrates, including endogenous substrates such as prostaglandin E2 and S-nitrosogluthathione and xenobiotic substrates such as anthracycline anti- cancer drugs (doxorubicin and daunorubicin) (15)(16)(17). Since doxorubicin causes DNA damage by intercalating itself into the DNA base pairs of the double helix, it is used for the treatment of broad range of solid tumors, including breast cancer and hepatocellular carcinoma cells (18,19).…”

Single nucleotide polymorphisms in ABCB1 and CBR1 can predict toxicity to R-CHOP type regimens in patients with diffuse non-Hodgkin lymphoma

“…Lars P. Jordheim, 1,2,3 Vincent Ribrag, 4 Hervé Ghesquieres, 5 Sophie Pallardy, 6 Richard Delarue, 7 Hervé Tilly,8 Corinne Haioun, 9 Fabrice Jardin, 10 Delphine Demangel, 2,3 Gilles A. Salles 11 …”

“…7 CBR1 has also been suspected to be involved in the occurrence of anthracycline-related toxicities as non-synonymous SNP were associated with reduced metabolism of doxorubicin and daunorubicin, and as these metabolisms were correlated to the expression of carbonyl reductases. 8,9 In addition, polymorphisms in CBR1 were correlated with altered pharmacokinetics with increased exposure to doxorubicin. 10 Finally, the cardioprotectant flavonoid 7-monohydroxyethyl rutoside was shown to behave as a CBR1 inhibitor.…”

Association between CBR1 polymorphisms and NSCLC in the Chinese population

“…CBR1 is also hypothesized to participate in cellular processes such as signal transduction (7), apoptosis (8), carcinogenesis (9) and drug resistance (10), and serves an important role in endometrial cancer (11), acute myeloid leukemia (12) and hepatocellular carcinoma (13). Schlager and Powis (14) reported that the level of CBR1 activity was significantly increased in lung tumors compared within normal tissue.…”