The Role of Cyp2b in Toxicant Metabolism and Sensitivity

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DESCRIPTION (provided by applicant): The cytochrome P450s (CYPs) are key enzymes involved in the detoxification of numerous chemicals including steroids, bile acids, pharmaceuticals, and environmental chemicals. Cyp2b is considered the forgotten CYP because not as much is known about its substrate specificity, it is expressed at lower levels in the liver than some other detoxification CYPs, and the FDA does not mandate the use of recombinant Cyp2b in drug metabolism testing. However, Cyp2b is highly inducible, highly polymorphic, gender predominant, and recent work shows it is expressed at higher levels than once thought, especially in hispanics. We will investigate the role of this CYP in the metabolism of environmental toxicants, determine whether individuals with low Cyp2b are sensitive to specific toxicants that are Cyp2b substrates, and evaluate whether CYP inhibition by toxicants could potentially lead to metabolic disorders such as obesity. We have recently developed a Cyp2b-knockdown mouse using RNAi that represses the expression of all five murine Cyp2b members. These mice demonstrate lower Cyp2b protein and activity and TCPOBOP the potent Cyp2b inducer is unable to out-compete the shRNA-mediated repression. In aim 1, we will use this mouse model to estimate Cyp2b metabolism using microsomes from untreated and TCPOBOP-treated wild-type and Cyp2b-KD mice. In aim 2, we will test whether Cyp2b-KD mice are susceptible to specific toxicants such as nonylphenol or parathion that are metabolized by Cyp2b. Last, recent preliminary data following ip injections with TCPOBOP or corn oil as a carrier revealed that Cyp2b-KD mice but not wild-type mice are unable to properly depurate the unsaturated fatty acids from the liver. This suggests an important role for Cyp2b in lipid homeostasis. In aim 3, we will test whether mice with low Cyp2b activity are susceptible to high-fat diet (Western diet) induced hepatic steatosis (fatty liver) and obesity. If Cyp2b-KD mice are susceptible to obesity, this would suggest that Cyp2b inhibitors may act as environmental obesogens. Overall, the purpose of this project is determine the role of Cyp2b in the metabolism, fate, and toxic effects of environmental toxicants in vivo using a novel murine model recently developed in our laboratory. PUBLIC HEALTH RELEVANCE: The cytochrome P450s (CYPs) are key enzymes involved in the detoxification of numerous chemicals including steroids, bile acids, pharmaceuticals, and environmental chemicals. Cyp2b is considered the forgotten CYP because it is expressed at lower levels in the liver than some other detoxification CYPs. However, it is highly inducible and recent work shows it is expressed at higher levels than once thought. We will investigate the role of this CYP in the metabolism of environmental toxicants, determine whether individuals with low Cyp2b are sensitive to specific toxicants, and evaluate whether CYP inhibition by toxicants could potentially lead to metabolic disorders.