Phenanthrene Metabolism in Smokers: Use of a Two-step Diagnostic Plot Approach to Identify Subjects with Extensive Metabolic Activation

Polycyclic aromatic hydrocarbons (PAHs) in cigarette smoke are among the most likely causes of lung cancer. PAHs require metabolic activation to initiate the carcinogenic process. Phenanthrene (Phe), a non-carcinogenic PAH, was used as a surrogate of benzo[α]pyrene (BaP) and related PAHs to study the metabolic activation of PAHs in smokers. A dose of 10 µg deuterated Phe ([D10]Phe) was administered to 25 healthy smokers in a crossover design, either as an oral solution or by smoking cigarettes containing [D10]Phe. Phe was deuterated to avoid interference from environmental Phe. Intensive blood and urine sampling was performed to quantitate the formation of deuterated r -1,t-2,3,c-4-tetrahydroxy-1,2,3,4-tetrahydrophenanthrene ([D10]PheT), a biomarker of the diol epoxide metabolic activation pathway. In both the oral and smoking arms approximately 6% of the dose was metabolically activated, with a large inter-subject variability in the formation of [D10]PheT observed. Two diagnostic plots were developed to identify subjects with large systemic exposure and significant lung contribution to metabolic activation, respectively. The combination of the two plots led to the identification of subjects with substantial local exposure. These subjects produced, in one single pass of [D10]Phe through the lung, a [D10]PheT exposure equivalent to the systemic exposure of a typical subject, and may be an indicator of lung cancer susceptibility. Polymorphisms in PAH metabolizing genes of the 25 subjects were also investigated. The integration of phenotyping and genotyping results indicated that GSTM1 null subjects produced approximately 2-fold more [D10]PheT than did GSTM1 positive subjects.