Nonalcoholic Fatty Liver Disease and Diabetes Are Associated with Decreased CYP3A4 Protein Expression and Activity in Human Liver

Abstract

Nonalcoholic fatty liver disease (NAFLD) is a major cause of chronic liver disease in the Western population. We investigated the association of nonalcoholic fatty liver disease (NAFLD) and diabetes mellitus on CYP3A4 activity in human liver tissue from brain dead donors ( n = 74). Histopathologically graded livers were grouped into normal ( n = 24), nonalcoholic fatty liver (NAFL, n = 26), and nonalcoholic steatohepatitis (NASH, n = 24) categories. The rate of conversion of midazolam to its 1-hydroxy metabolite was used to assess in vitro CYP3A4 activity in human liver microsomes (HLM). A proteomics approach was utilized to quantify the protein expression of CYP3A4 and related enzymes. Moreover, a physiologically based pharmacokinetic (PBPK) model was developed to allow prediction of midazolam concentration in NAFL and NASH livers. CYP3A4 activity in NAFL and NASH was 1.9- and 3.1-fold ( p < 0.05) lower than normal donors, respectively. Intrinsic clearance (CLint) was 2.7- ( p < 0.05) and 4.1-fold ( p < 0.01) lower in donors with NAFL and NASH, respectively. CYP3A4 protein expression was significantly lower in NAFL and NASH donors ( p < 0.05) and accounted for significant midazolam hydroxylation variability in a multiple linear regression analysis (β = 0.869, r = 0.762, P < 0.01). Diabetes was also associated with decreased CYP3A4 activity and protein expression. Both midazolam CLint and CYP3A4 protein abundance decreased significantly with increase in hepatic fat accumulation. Age and gender did not exhibit any significant association with the observed alterations. Predicted midazolam exposure was 1.7- and 2.3-fold higher for NAFL and NASH, respectively, which may result in a longer period of sedation in these disease-states. Data suggests that NAFLD and diabetes are associated with the decreased hepatic CYP3A4 activity. Thus, further evaluation of clinical consequences of these findings on the efficacy and safety of CYP3A4 substrates is warranted.

Quantitative Proteomics and Mechanistic Modeling of Transporter‐Mediated Disposition in Nonalcoholic Fatty Liver Disease

“…About 30% of total morphine metabolism is thought to be mediated by CYP3A . NASH‐induced reduction of CYP3A expression and function (~ 50–70%) may increase morphine glucuronide formation in NASH. However, PBPK analysis implied such changes (captured in the current model) may only partly explain the clinical observation.…”

Non-alcoholic Steatohepatitis (NASH) Drug Discovery – Building a Consensus on ADME Screening Tools and Clinical Pharmacology Strategies to Aid Candidate Development

“…The CYP3A4 expression in diabetes+NAFLD and diabetes+NASH was 4.2 and 2.9-fold lower as compared to normal diabetic liver (26).…”

“…The CYP3A4 activity in livers from diabetic donors with NAFLD and NASH was 4.5 and 5.7-fold lower as compared to the normal diabetic liver. The intrinsic clearance in diabetic livers with NAFLD and NASH was 1.4-fold lower as compared to diabetic liver without fatty disease (26).…”

“…The results showed a 1.9 and 3.1-fold decrease in CYP3A4 activity in NAFLD and NASH, respectively. Also, the intrinsic clearance was 2.7 and 4.1-fold lower in NAFLD and NASH liver donors, respectively with an overall decrease in CYP3A4 expression by 2-fold in both NAFLD and NASH (26). The CYP3A4 activity in livers from diabetic donors with NAFLD and NASH was 4.5 and 5.7-fold lower as compared to the normal diabetic liver.…”

“…Na et al demonstrated from in vitro and in vivo Cyp cocktail assays, Cyp1A, Cyp2B, Cyp2C and Cyp3A were significantly decreased, whereas Cyp2D remained unchanged in MCD diet mice as compared to normal mice (25). Jamwal et al (2018) studied the association of NAFLD, NASH and diabetes mellitus on CYP3A4 activity in human liver tissue from brain dead donors (26). The results showed a 1.9 and 3.1-fold decrease in CYP3A4 activity in NAFLD and NASH, respectively.…”