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Principal Investigator: FRANK J GONZALEZ
Organization: DIVISION OF BASIC SCIENCES - NCI
Fiscal Year: 2020
Award: $866,441
Funding agency: National Cancer Institute
Nonalcoholic steatohepatitis (NASH) is the progressive stage of nonalcoholic fatty liver disease that highly increases the risk of cirrhosis and liver cancer, and there are few therapeutic options available in the clinic. While virus-induced hepatitis has been sharply reduced with vaccine application and curative drugs, there continues to be an increase in NAFLD incidence due to the rapid rise of obesity and diabetes. NASH is currently listed as the second leading cause of liver disease among adults awaiting liver transplantation in the United States and is estimated to overtake hepatitis C virus infection as the leading cause of liver transplantation in the United States in the coming decades. Withaferin A (WA), extracted from the ayurvedic medicine Withania somnifera, has a wide range of pharmacological activities; however, little is known about its effects on NASH. To explore the role of WA in treating NASH, two well defined NASH models were used, the methionine-choline-deficient diet and the 40 kcal% high-fat diet (HFD). In both NASH models, WA treatment or control vehicle was administered to evaluate its hepatoprotective effects. As assessed by biochemical and histologic analyses, WA prevented and therapeutically improved liver injury in both models, as revealed by lower serum aminotransaminases, hepatic steatosis, liver inflammation, and fibrosis. In the HFD-induced NASH model, both elevated serum ceramides and increased hepatic oxidative stress were decreased in the WA-treated group compared with the control vehicle-treated group. To further explore whether WA has an anti-NASH effect independent of its known action in leptin signaling associated with obesity, leptin signaling-deficient ob/ob mice maintained on an HFD were used to induce NASH. WA therapeutically reduced NASH in HFD-treated leptin-deficient ob/ob mice, thus demonstrating a leptin-independent hepatoprotective effect. This study revealed that WA treatment could be an option for NASH treatment and prevention of hepatocellular carcinogenesis. Inflammatory bowel diseases (IBD) are chronic relapsing gastrointestinal disorders, commonly exemplified by ulcerative colitis or Crohn's disease. IBD pathogenesis is closely related to life style, genetic, and environmental etiological factors, while chronic IBD presents a high risk for colorectal cancer both in mice and humans. Globally, an increasing incidence of IBD has been found over the past decades and IBD has added economic and psychological burden for patients. Although numerous therapeutic advances have emerged in the clinic in recent years, no drugs are available to cure this refractory disease, and thus novel therapeutic choices are urgently needed. Rutaecarpine (RUT), isolated from the Traditional Chinese Medicine (TCM) of Evodia rutaecarpa, was reported to suppress IBD. However, the mechanism by which RUT ameliorates dextran sulfate sodium (DSS)-induced IBD is largely unknown. By use of nuclear factor-erythroid 2-related factor 2 (NRF2) knockout mice, cell-based studies, surface plasmon resonance (SPR), western blotting analysis, and molecular docking studies, the mechanism by which RUT affects DSS-induced colitis was explored. In DSS-treated wild-type mice but not in Nrf2-null mice, RUT significantly improved colitis as revealed by rescued body weight loss, improved histology and inflammation, and induced expression of NRF2 target genes in colon and ileum. Cell-based studies showed that RUT significantly increased the LD50 for hydrogen peroxide (H2O2)-induced cell damage, activated NRF2 nuclear translocation, and suppressed the production of reactive oxygen species in H2O2-treated HCT116 cells, activated NRF2 luciferase reporter activities in HCT116 cells and HepG2 cells, and induced expression of NRF2 target genes in primary intestinal epithelial cells. Molecular docking in silico and SPR assays indicated that RUT interacted with kelch-like ECH-associated protein 1 (KEAP1), and extracellular incubation studies revealed that RUT bound to the KEAP1 kelch domain with a calculated equilibrium dissociation constant Kd of 19.6 uM. In conclusion, these results demonstrate that RUT ameliorates DSS-induced colitis, dependent on NRF2, and could be a potential therapeutic option for IBD patients. Mechanistically, RUT potentiates NRF2 nuclear translocation to upregulate NRF2-mediated antioxidant response by directly inhibiting KEAP1-NRF2 interaction. The cytochrome P450 2D (CYP2D) subfamily belongs to the cytochrome P450-dependent mixed function oxidase system that includes several major drug-metabolizing enzymes. CYP2D isoforms are expressed in various tissues, such as the liver, intestine, kidney and brain and, in humans, catalyze the hepatic metabolism of about 25% of the prescribed drugs including the vast majority of antidepressants and antipsychotics, several analgesics, antiarrythmics and anticancer drugs. CYP2D is also implicated in the synthesis and metabolism of neurotransmitters (dopamine and serotonin) and neurosteroids in the human and rat brain and mainly in brain regions where disturbances in these neurotransmitter systems contribute to the pathophysiology of depression and schizophrenia. It is also noteworthy that CYP2D6 polymorphisms have been implicated in personality traits and neurological or psychiatric disorders; reduced CYP2D6 activity is associated with increased susceptibility to Parkinson's disease. Interestingly, the reduced serotonin metabolism in CYP2D6 poor metabolizers (PMs) has been associated with a cluster of behavioral traits such as anxiety, impulsivity and attention deficits. However, the precise role of CYP2D6 in brain physiology remains an enigma. The CYP2D subfamily catalyses the metabolism of about 25% of prescribed drugs, including the majority of antidepressants and antipsychotics. At present, the mechanism of hepatic CYP2D regulation remains largely unknown. This study investigated the role of sex steroid hormones in CYP2D regulation. For this purpose, Cyp2d22 expression was assessed in the distinct phases of the estrous cycle of normocyclic C57BL/6J (WT) female mice. Cyp2d22 was also evaluated in ovariectomised WT and CYP2D6-humanized (hCYP2D6) mice that received hormonal supplementation with either 17beta-estradiol (E2) and/or progesterone. Comparisons were also made to male mice. The data revealed that hepatic Cyp2d22 mRNA, protein and activity levels were higher at estrous compared to the other phases of the estrous cycle and that ovariectomy repressed Cyp2d22 expression in WT mice. Tamoxifen, an anti-estrogenic compound, also repressed hepatic Cyp2d22 via activation of GH/STAT5b and PI3k/AKT signaling pathways. Both hormones prevented the ovariectomy-mediated Cyp2d22 repression. In case of progesterone, this may be mediated by inhibition of the PI3k/AKT/FOX01 pathway. Notably, Cyp2d22 mRNA levels in WT males were similar to those in ovariectomised mice and were markedly lower compared to females at estrous, a differentiation potentially regulated by the GH/STAT5b pathway. Sex steroid hormone-related alterations in Cyp2d22 mRNA expression were highly correlated with Hnf1a mRNA. Interestingly, fluctuations in Cyp2d22 in hippocampus and cerebellum followed those in liver. In contrast to WT mice, ovariectomy induced hepatic CYP2D6 expression in hCYP2D6 mice, whereas E2 and/or progesterone prevented this induction. Apparently, sex steroid hormones display a significant gender- and species-specific role in the regulation of CYP2D.
Terms: <2-amino-ethanesulfonic acid><21+ years old><5-HT><5-Hydroxytryptamine><5HT><AKT><AKT Signaling Pathway><Abnormal Assessment of Metabolism><Active Oxygen><Adult><Adult Human><Adult-Onset Diabetes Mellitus><Affect><Akt protein><Amino Acids><Aminoacetic Acid><Ammon Horn><Analgesic Agents><Analgesic Drugs><Analgesic Preparation><Analgesics><Anodynes><Anti-Cancer Agents><Anti-Estrogens><Antidepressant Agent><Antidepressant Drugs><Antidepressants><Antidepressive Agents><Antineoplastic Agents><Antineoplastic Drugs><Antineoplastics><Antinociceptive Agents><Antinociceptive Drugs><Antioxidants><Antipsychotic Agents><Antipsychotic Drugs><Antipsychotics><Anxiety><Aquadiol><Area><Ashwagandha><Assay><Attentional deficit><Autoregulation><Ayurvedic Medicine><Back><Behavioral><Bile Acid Biosynthesis><Bile Acid Biosynthesis Pathway><Bile Acids><Bioassay><Biochemical><Biologic Assays><Biological><Biological Assay><Biological Markers><Blood><Blood Circulation><Blood Reticuloendothelial System><Blood Serum><Bloodstream><Body Tissues><Body Weight decreased><Brain><Brain Nervous System><Brain region><Breast Neoplasms><Breast Tumors><CCR><CPD6><CYP 2D6><CYP2D><CYP2D6><CYP2D6 gene><CYP2DL1><CYPIID6><Cancer Causing Agents><Cancer Control><Cancer Control Science><Cancer Detection><Cancer Drug><Cancer Model><Cancer Patient><Cancer Screening for Patients><Cancer Treatment><CancerModel><Cancers><Carcinogen Metabolism><Carcinogens><Case-Base Studies><Case-Comparison Studies><Case-Compeer Studies><Case-Control Studies><Case-Referent Studies><Case-Referrent Studies><Catabolism><Causality><Cell Body><Cell Communication and Signaling><Cell Signaling><Cells><Cellular injury><Ceramides><Cerebellum><Chemicals><Chemoprotection><Chinese Herbs><Chinese Traditional Medicine><Chinese medicine><Cholesterol><Chronic><Chung I Hsueh><Circulation><Cirrhosis><Clinic><Clinical><Colitis><Colon><Colon Cancer><Colon Carcinoma><Colonic Carcinoma><Common Rat Strains><Complex Mixtures><Cornu Ammonis><Corpus Luteum Hormone><Crohn disease><Crohn's><Crohn's disease><Crohn's disorder><Cytochrome P-450><Cytochrome P-450 CYP2D6><Cytochrome P-450 Enzyme System><Cytochrome P450><Cytochrome P450 2D6><Cytochrome P450 Family Gene><Cytochrome P450 Subfamily IID Polypeptide 6><DSS-induced colitis><Data><Debrisoquine 4-Hydroxylase><Debrisoquine 4-Monooxygenase><Debrisoquine Hydroxylase><Delta4-pregnene-3,20-dione><Diabetes Mellitus><Diagnostic><Dimenformon><Diogyn><Diogynets><Discontinuous Capillary><Disease><Disorder><Dissociation><Distal><Docking><Dopamine><Dorsum><Drug Prescribing><Drug Prescriptions><Drugs><Dysfunction><Economics><Encephalon><Endocrine Gland Secretion><Enteramine><Enzyme Gene><Enzymes><Epithelial Cells><Equilibrium><Estrace><Estradiol><Estradiol-17 beta><Estradiol-17beta><Estraldine><Estrogen Antagonists><Estrous Cycle><Estrus><Etiology><Euodia ruticarpa><Evodia rutaecarpa><Evodiae Fructus><Fatty Liver><Female><Fibrosis><Functional disorder><Gall Bladder><Gallbladder><Gallbladder / Biliar><Gallbladder/Biliary system><Gastrointestinal Diseases><Gender><Genes><Genetic><Genetic Models><Genetic Polymorphism><Glycine><Gonadal Steroid Hormones><Granulomatous Enteritis><H2O2><HCT 116 Cells><HCT- 116><HCT-116><HCT116><HCT116 Cells><HCV infection><Hep G2><HepG2><HepG2 cell line><Hepatic><Hepatic Cancer><Hepatic Disorder><Hepatic Transplantation><Hepatitis><Hepatitis C><Hepatitis C virus infection><Hepatitis, Viral, Non-A, Non-B, Parenterally-Transmitted><Hepatitus C><Hepatocarcinogenesis><Herbal Medicine><High Fat Diet><Hippocampus><Hippocampus (Brain)><Hippophaine><Histologic><Histologically><Histology><Homeostasis><Hormonal><Hormones><Human><Hydrogen Peroxide><Hydroperoxide><Hydroxylases><Hydroxytyramine><Hyperlipemia><Hyperlipidemia><Imipramine 2-Hydroxylase><Impulsivity><Incidence><Individual><Inflammation><Inflammatory Bowel Diseases><Inflammatory Bowel Disorder><Inflammatory Intestinal Disease><Inflammatory Intestinal Disorder><Injury to Liver><Insulin Resistance><Intermediary Metabolism><Intestinal><Intestines><Intracellular Communication and Signaling><Investigators><Isoforms><KO mice><Ketosis-Resistant Diabetes Mellitus><Kidney><Kidney Urinary System><Knock-out Mice><Knockout Mice><LD-50><LD50><Lead><Leptin><Lethal Dose 50><Life Style><Lifestyle><Lipid Trafficking><Lipids><Liver><Liver Carcinogenesis><Liver Grafting><Liver Steatosis><Liver Transplant><Liver diseases><Luciferase Immunologic><Luciferases><Major Tranquilizers><Major Tranquilizing Agents><Malignant Neoplasm Therapy><Malignant Neoplasm Treatment><Malignant Neoplasms><Malignant Tumor><Malignant Tumor of the Lung><Malignant neoplasm of liver><Malignant neoplasm of lung><Mammary Cancer><Mammary Neoplasms><Maps><Maturity-Onset Diabetes Mellitus><Mediating><Medication><Mental Depression><Mental disorders><Mental health disorders><Messenger RNA><Metabolic><Metabolic Diseases><Metabolic Disorder><Metabolic Processes><Metabolic Studies><Metabolism><Metabolism Studies><Methionine><Methodology><Mice><Mice Mammals><Minor><Mixed Function Oxidases><Mixed Function Oxygenases><Modeling><Modern Man><Molecular><Monitor><Monooxygenases><Murine><Mus><NAFLD><NASH><NF-E2 protein><NF-E2 transcription factor><NFE2 protein><NIDDM><Natural Products><Neoplastic Disease Chemotherapeutic Agents><Nerve Transmitter Substances><Nervous System Diseases><Neuroleptic Agents><Neuroleptic Drugs><Neuroleptics><Neurologic Disorders><Neurological Disorders><Neurotransmitters><Non-Insulin Dependent Diabetes><Non-Insulin-Dependent Diabetes Mellitus><Noninsulin Dependent Diabetes><Noninsulin Dependent Diabetes Mellitus><Nuclear Translocation><Null Mouse><Ob Gene Product><Ob Protein><Obese Gene Product><Obese Mice><Obese Protein><Obesity><Oncogens><Oophorectomy><Organism><Ovariectomy><Ovocyclin><Ovocylin><Oxidative Stress><Oxygen Radicals><P450><P450-2D6><P450-DB1><P450C2D><P450DB1><PI3CG><PI3KGamma><PI3k><PIK3><PIK3CG><PIK3CG gene><PPAR alpha><PPAR-α><PPARalpha><PPARα><Paralysis Agitans><Parkinson><Parkinson Disease><Parkinson's disease><Parkinsons disease><Pathogenesis><Pathway interactions><Patients><Pb element><Peroxisome Proliferator-Activated Receptor alpha><Peroxisome Proliferator-Activated Receptor α><Personality Traits><Pharmaceutic Preparations><Pharmaceutical Preparations><Pharmacology><Phase><Physiological Homeostasis><Physiology><Physiopathology><Portal Vein><Portal vein structure><Predisposition><Pregn-4-ene-3,20-dione><Pregnenedione><Prevention><Primary Parkinsonism><Pro-Oxidants><Production><Progesterone><Progynon><Protein Isoforms><Protein Kinase B><Proteins><Proto-Oncogene Proteins c-akt><Psychiatric Disease><Psychiatric Disorder><Pulmonary Cancer><Pulmonary malignant Neoplasm><RAC-PK protein><Rat><Rats Mammals><Rattus><Reaction><Reactive Oxygen Species><Receptor Protein><Refractory Disease><Regulation><Relapse><Reporter><Reporting><Repression><Research Personnel><Researchers><Risk><Role><Sampling><Schizophrenia><Schizophrenic Disorders><Screening for cancer><Serotonin><Serum><Sex Hormones><Sex Steroid Hormones><Signal Transduction><Signal Transduction Systems><Signaling><Sinusoid><Sinusoidal Capillary><Slow-Onset Diabetes Mellitus><Small Intestines><Sodium Dextran Sulfate><Sparteine Monooxygenase><Stable Diabetes Mellitus><Subfamily IID Cytochrome P450><Subfamily IID-Like 1 Cytochrome P450><Supplementation><Surface Plasmon Resonance><Susceptibility><System><T2 DM><T2D><T2DM><Tamoxifen><Tauphon><Taurine><Technology><Tetradium ruticarpum><Therapeutic><Therapeutic Estradiol><Therapeutic Hormone><Therapeutic Progesterone><Thesaurismosis><Tissues><Toxic effect><Toxicities><Tumor-Specific Treatment Agents><Type 2 Diabetes Mellitus><Type 2 diabetes><Type II Diabetes Mellitus><Type II diabetes><Ulcerated Colitis><Ulcerative Colitis><United States><Urine><Urine Urinary System><Vaccines><Virus><Weight Loss><Weight Reduction><Western Blotting><Western Immunoblotting><Wild Type Mouse><Withania somnifera><Xenobiotic Metabolism><Xenobiotics><Zhong Yi Xue><adiposity><adult onset diabetes><adulthood><aminoacid><anti-cancer drug><anti-cancer therapy><anti-depressant agent><anti-depressant drugs><anti-depressants><anti-depressive agents><anti-oxidant><anticancer agent><anticancer drug><anticancer therapy><antiestrogen><antiestrogenic><ayurveda><ayurvedic><bacteria in the gut><balance><balance function><base><bile acid anabolism><bile acid biosynthetic process><bile acid formation><bile acid metabolism><bile acid synthesis><bile duct><bile ductule><bile metabolism><bio-markers><biologic marker><biological signal transduction><biomarker><biomarker discovery><body weight loss><bowel><c-akt protein><cancer biomarkers><cancer chemoprevention><cancer in the colon><cancer markers><cancer recurrence><cancer therapy><carcinogenesis in the liver><carcinogenicity><causation><cell damage><cell injury><cellular damage><chemical carcinogen><chinese herbal><choline deficient diet><cirrhotic><colitis-induced dysbiosis><colo-rectal cancer risk><colorectal cancer risk><corpulence><corpulency><corpulentia><damage to cells><dementia praecox><depression><dextran sulfate sodium induced colitis><diabetes><disease causation><drug metabolism><drug/agent><early biomarkers><early cancer detection><early detection biomarkers><early detection markers><eleocolitis><estrogen inhibitor><estrous><extracellular><female gonadectomy><gastrointestinal disorder><gonadal steroids><gut bacteria><heavy metal Pb><heavy metal lead><hep C><hepatic body system><hepatic carcinogenesis><hepatic damage><hepatic disease><hepatic injury><hepatic metabolism><hepatic organ system><hepatic steatosis><hepatitis non A non B><hepatocellular carcinogenesis><hepatopathy><high risk><hippocampal><humanized mice><humanized mouse><ileum><improved><in silico><injury to cells><instrument><insulin resistant><intestinal epithelium><ketosis resistant diabetes><knockout gene><lipid transport><liver cancer><liver cancer pathogenesis><liver damage><liver disorder><liver inflammation><liver injury><liver metabolism><liver transplantation><liver tumorigenesis><living system><lung cancer><mRNA><mRNA Expression><male><malignancy><malignant liver tumor><mammary tumor><maturity onset diabetes><mental illness><metabolic abnormality assessment><metabolism disorder><metabolism measurement><metabolomics><metabonomics><mouse model><murine model><neoplasm/cancer><nervous system disorder><neuroactive steroids><neurological disease><neurosteroids><neurotransmitter metabolism><new drug treatments><new drugs><new therapeutics><new therapy><next generation therapeutics><non A, non B hepatitis><non-A, non-B hepatitis><non-alcohol fatty liver disease><non-alcohol induced steatohepatitis><non-alcoholic fatty liver disease><non-alcoholic liver disease><non-alcoholic steato-hepatitis><non-alcoholic steatohepatitis><nonalcoholic fatty liver disease><nonalcoholic steato-hepatitis><nonalcoholic steatohepatitis><novel><novel drug treatments><novel drugs><novel therapeutics><novel therapy><nuclear factor-erythroid 2><ob/ob mouse><obese><obese people><obese person><obese population><oncogenic agent><oxidation><pain killer><pain medication><pain reliever><painkiller><pathophysiology><pathway><polymorphism><prevent><preventing><programs><protein blotting><proto-oncogene protein RAC><proto-oncogene protein akt><psychiatric illness><psychologic><psychological><psychological disorder><rac protein kinase><receptor><regional enteritis><related to A and C-protein><renal><response><schizophrenic><sex steroid><small bowel><social role><trait><type 2 DM><type II DM><type two diabetes><uptake><wildtype mouse><wt-loss>