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Edit Comment Close Premium member Presentation Transcript The Endocannabinoid System and Its Role in Obesity and Cardiovascular Disease: The Endocannabinoid System and Its Role in Obesity and Cardiovascular Disease Discovery Timeline of the Endocannabinoid System: Discovery Timeline of the Endocannabinoid System 2600 BC – Chinese emperor Huang Ti advised taking Cannabis for relief of cramps, and rheumatic and menstrual pain 1839 – Publication by British physician O'Shaughnessy publicizes therapeutic potential of Cannabis to Western world 1964 – Isolation of 9-THC, the active constituent of Cannabis sativa 1991 – Cloning of the human CB1 receptor 1988 – First identification of cannabinoid receptors in rat brain 1990 – Cloning of the first G protein- coupled cannabinoid receptor, CB1 in rat Discovery Timeline (cont’d): 1992 – Discovery of anandamide, the first endogenous cannabinoid 1993 – Cloning of the CB2 receptor 1994 – Characterization of the first CB1 receptor inhibitor, SR141716 (rimonabant) 1995 – 'Endocannabinoid' coined by Di Marzo 1995 – Isolation of a second endocannabinoid, 2-AG 2000s – Wide tissue distribution of CB1 receptors Characterization of peripheral ECS Role of ECS in human obesity and obesity-related metabolic disorders Discovery Timeline (cont’d) Foundations of Endocannabinoid Biology: Foundations of Endocannabinoid Biology Receptors Ligands Pathways of synthesis and degradation Signaling Introduction to physiological effects 4 Slide5: I. The Endocannabinoid System (ECS): Receptors CB1 are located in several areas of the brain and in a variety of tissues such as adipose tissue, liver, muscle, the gastrointestinal tract, pancreas, gonad and sensory neurons CB2 receptors are located in immune cells such as T-cells T-cells B-cells Monocytes Spleen Tonsils 5 Howlett AC et al., Pharmacol Rev. 2002, 54: 161-202. Slide6: II. The ECS: Ligands that Bind to Cannabinoid Type 1 (CB1) Receptors Plant-derived Endogenously produced 3 types: endogenously produced, plant-derived, and synthetic Endogenous ligands (endocannabinoids) synthesized from membrane-derived phospholipids As lipophilic compounds, endocannabinoids are not stored within the synaptic vesicle cytoplasm, but can diffuse freely across cell membranes Di Marzo V, et al. Nat Neurosci. 2005;8:585-589. 6 D9-Tetrahydrocannabinol Slide7: III. The ECS: Pathways of Ligand Synthesis and Degradation Endocannabinoids are produced on demand from phospholipid-derived precursors in the cell membrane Anandamide and 2-AG act as retrograde messengers Fatty Acid Amide Hydrolase rapidly degrades these ligands 7 Freund TF et al. Physiol Rev. 2003, 83: 1017-1066. Classical neurotransmitter: Acetylcholine (Ach): Classical neurotransmitter: Acetylcholine (Ach) IV. The ECS Signaling: Contrast Between Classical Neurotransmission and the ECS 8 Di Marzo V, et al. Nat Rev Drug Discov. 2004;3:771-784. Slide9: Di Marzo V, et al. Nat Rev Drug Discov. 2004;3:771-784. V. The ECS: Cellular Effects of CB1 Receptor Activation 9 Gaps in Current Understanding of the Endocannabinoid System: Gaps in Current Understanding of the Endocannabinoid System How are endocannabinoids taken back up into cells? What is the relevance of circulating endocannabinoid levels? If they are produced on demand and degraded immediately, how can circulating levels be measured? How do endocannabinoids act in peripheral tissues? Where do they come from? Are peripheral effects dependent on neuronal innervation? 10 Gaps in Current Understanding of the Endocannabinoid System: Gaps in Current Understanding of the Endocannabinoid System What are the relative contributions of central and peripheral endocannabinoid system activities to the overall clinical impact of CB1 blockade? To what extent are metabolic effects of CB1 blockade dependent on concomitant reductions in food intake and body weight, and to what degree are they attributable to direct pharmacological effects? 11 Some CB1 Receptor Antagonists Currently in Development: Some CB1 Receptor Antagonists Currently in Development Phase 1 Solvay Pharmaceuticals, Bristol-Myers Squibb SLV 319 Phase 2,3 Merck L-000899055 (MK0364) Phase 1 Pfizer CP-945598 Under FDA review Sanofi-Aventis Rimonabant (SR141716) andamp; SR141778 Stage of Development Manufacturer Drug 12 The ECS Modulates:: The ECS Modulates: 13 Energy Balance Feeding Behavior Hepatic Lipogenesis Glucose Homeostasis Sites of Action for CB1 Modulation of Energy Homeostasis: Energy Balance Feeding Behavior Gastric emptying GI motility Hepatic glucose output Lipogenesis Hepatic Lipogenesis Adipose Tissue Metabolism Glucose Homeostasis Glucose uptake Glucose, lipid oxidation Lipolysis Lipogenesis Ghrelin, PYY Limbic forebrain Motivation for palatable food Sites of Action for CB1 Modulation of Energy Homeostasis 14 Slide15: Energy balance EC blockade reduces food intake and body weight CB1 deletion is associated with a lean phenotype and resistance to diet-induced obesity Feeding behavior Stimulation of CB1 receptors increases food intake whereas CB1 blockade reduces food intake Supporting Evidence: 15 Wiley JL et al. Br J Pharmacol. 2005;145:293-300. Cota D et al. J Clin Invest. 2003;112(3):423–431. Hao S et al. Eur J Pharmacology. 2000; 392:147-156. Endocannabinoids Stimulate Food Intake in Mice: Endocannabinoids Stimulate Food Intake in Mice Hao S et al. Eur J Pharmacology. 2000; 392:147-156. Anandamide 0.001 mg/kg Vehicle *Pandlt;0.05; **Pandlt;0.01 vs vehicle 1 3 5 7 Day Food intake (grams/day) 7 6 5 4 3 * * * ** 16 CB1 Blockade Produces a Dose-Related Reduction in Food Intake in Mice: CB1 Blockade Produces a Dose-Related Reduction in Food Intake in Mice Wiley JL et al. Br J Pharmacol. 2005;145:293-300. 2.0 1.5 1.0 0.5 0.0 0.0 0.3 1.0 3.0 10.0 Rimonabant Dose (mg/kg-1) Food intake (g) 17 Slide18: CB1 Knockout (CB1-/- ) Mice Sustain a Reduced Body Weight vs. Pair-fed Wildtype Mice Cota D et al. J Clin Invest. 2003;112(3):423–431. Pair-feeding showed that activation of metabolic processes other than reduced food intake contribute to the lean phenotype in CB1 knockout mice 20 21 22 15 20 25 30 35 40 45 Age (week) Body weight (g) 18 CB1+/+ WILDTYPE CB1+/+ WILDTYPE CB1-/- KNOCKOUT Pair-fed Slide19: Hepatic lipogenesis High fat diet and CB1 activation increase hepatic SREBP-1c and the associated lipogenic enzymes FAS and ACC CB1 blockade attenuates these changes and the fatty liver that occurs as a consequence CB1 knockout animals are resistant to increased hepatic SREBP-1c and hepatic FAS and ACC expression as well as the resulting fatty liver Supporting Evidence: 19 Osei-Hyiaman D, et al. J. Clin. Invest. 2005;115:1298-1305. The ECS is Overactivated in Liver of Diet-Induced Obese Animals, Leading to Fatty Liver in Wildtype but not CB1 Knockout Animals: The ECS is Overactivated in Liver of Diet-Induced Obese Animals, Leading to Fatty Liver in Wildtype but not CB1 Knockout Animals Osei-Hyiaman D, et al. J. Clin. Invest.2005;115:1298-1305. Hepatic anandamide (AEA) levels are elevated in diet-induced obesity, more in wildtype vs CB1 knockout animals AEA (fmol/mg tissue) 20 Slide21: A. CB1 activation increases while CB1 blockade reduces expression of lipogenic enzymes Osei-Hyiaman D, et al. J. Clin. Invest. 2005;115:1298-1305. 21 Vehicle CB1 CB1 CB1 agonist + agonist blockade blockade 300 200 100 0 200 100 0 200 100 0 * * SREBP-1c mRNA (% of control) FAS mRNA (% of control) ACC1 mRNA (% of control) *Pandlt;0.05 vs Vehicle Fatty acid synthase Acetyl CoA carboxylase Hepatic Lipogenesis is Modulated by CB1 Activation Slide22: Osei-Hyiaman D, et al. J. Clin. Invest. 2005;115:1298-1305. 22 Rate of fatty acid synthesis (mmol/g/h) 80 40 0 Normal diet High fat diet Vehicle Rimonabant Vehicle Rimonabant * B. De novo hepatic fatty acid synthesis is increased by high-fat diet through CB1 receptor activation and is attenuated by CB1 blockade Hepatic Lipogenesis is Modulated by CB1 Activation *P andlt; 0.01 vs. vehicle Slide23: Glucose homeostasis After high fat feeding, CB1 knockout animals have a lower glucose response to insulin injection than wild-type animals Administration of CB1 blockade for 7 days to obese animals leads to increased insulin-stimulated glucose uptake by isolated soleus muscle Supporting Evidence: 23 Ravinet Trillou C et al. Int J Obes. 2004;28:640–648. Liu YL et al. Int J Obes Relat Metab Disord. 2005;29:183-187. Slide24: CB1 Knockout Mice Are Leaner and Resistant to Diet-Induced Obesity and Its Metabolic Consequences Ravinet Trillou C et al. Int J Obes. 2004;28:640–648. **Pandlt;0.01 vs CB+/+ on standard diet Standard High fat Glucose response to intraperitoneal (IP) insulin 200 150 100 50 0 ** Glucose AUC 0-180 min Glycemia AUC from 0-180 min after i.p. insulin (0.6 U/kg) injection CB1-/- Knockout CB1+/+ Wildtype 24 Slide25: Effect of 7-day treatment with Rimonabant (10 mg/kg; i.p. once daily) on glucose uptake (formation of 2-deoxy[3H]-glucose-6-phosphate) in isolated soleus muscle in the presence of 10 nM insulin CB1 Receptor Blockade Stimulates Muscle Glucose Uptake in Obese Rats Muscle glucose uptake Liu YL et al. Int J Obes Relat Metab Disord. 2005;29:183-187. 25 Slide26: Adipose tissue metabolism EC stimulation with CB1 agonist increases adipose tissue LPL expression while CB1 blockade inhibits this effect CB1 stimulation reduces while blockade increases adiponectin synthesis CB1 blockade reverses the histological changes in adipose tissue produced by diet-induced obesity EC stimulation reduces the expression of AMP kinase in visceral fat Supporting Evidence: 26 Cota D et al. J Clin Invest. 2003;112:423. Matias I, et al. XV ICRS Symposium June 24-27, 2005; Clearwater, Fla. Jbilo O, et al. FASEB J. 2005;19:1567-1569. Slide27: The Peripheral ECS in Adipose Tissue Cota D et al. J Clin Invest. 2003;112:423. * Pandlt;0.05 vs vehicle control † Pandlt;0.05 vs CB1 agonist Rimonabant inhibits the increase in lipoprotein lipase expression produced by CB1 stimulation 300 200 100 0 LPL activity (% of control) - + + - - - + + CB1 agonist Rimonabant * † † 27 Slide28: The Peripheral ECS in Adipose Tissue Matias I, et al. XV ICRS Symposium June 24-27, 2005; Clearwater, Fla. CB1 stimulation significantly reduced, while rimonabant increased, adiponectin. 28 Relative Adiponectin Expression on day 12 ** ** Control CB1 CB1 stimulation blockade (HU-210) (Rimonabant) **Pandlt;0.01 vs Control Slide29: The Peripheral ECS in Adipose Tissue Adipose tissue of obese mice fed a high fat diet (HFD) plus rimonabant resembles that of lean mice fed a standard diet (STD) Jbilo O, et al. FASEB J. 2005;19:1567-1569. 29 The ECS is Overactivated in:: The ECS is Overactivated in: 30 Animal models of genetic obesity Animal models of diet-induced obesity Human obesity Supporting Evidence: Supporting Evidence Animal models of genetic obesity Increased endocannabinoid levels in several genetic obesity models Increased endocannabinoid levels and CB1 receptor expression in diet-induced obesity 31 Bensaid M, et al. Mol Pharm. 2003;63:908-914. The ECS Is Overactivated in Adipose Tissue of Genetically Obese Rats: The ECS Is Overactivated in Adipose Tissue of Genetically Obese Rats Bensaid M, et al. Mol Pharm. 2003;63:908-914. 32 CB1 Receptor mRNA Level (Arbitrary Units) 0 1 2 3 4 5 Lean Obese Rats U 3T3 F442A Cells 0 1 2 3 4 5 0 1 2 3 4 5 D Up-regulation of CB1 receptor mRNA In white adipose tissue of obese (fa/fa) vs. lean Zucker rats) In mouse 3T3 F442A adipocyte cells Slide33: 10 8 6 4 2 0 16 14 12 10 8 5 4 18.5-24.9 25.0-29.9 andgt;30.0 BMI (kg/m2) 18.5-24.9 25.0-29.9 andgt;30.0 Caucasians African Americans Pandlt;0.05* Pandlt;0.01* Pandlt;0.05* BMI (kg/m2) Percent of subjects with FAAH 385 A/A genotype by BMI category % of subjects with FAAH 385 A/A * vs normal BMI A Mutation in the Enzyme That Degrades Endocannabinoids is Associated with Increased BMI Sipe JC et al. Int J Obes Relat Metab Disord.2005;29:755-759. 33 Slide34: Engeli S, et al. Diabetes 2005; 54:2838–2843. * Pandlt;0.05 vs lean women The ECS is Upregulated in Human Obesity 34 Supporting Evidence: Supporting Evidence The endocannabinoid system is up-regulated in human obesity Missense mutation in FAAH, the primary enzyme involved in endocannabinoid degradation, is associated with increased BMI Circulating endocannabinoid levels are increased, while FAAH is reduced in obese women FAAH expression in adipose tissue is reduced in obese vs. lean women 35 Sipe JC et al. Int J Obes Relat Metab Disord.2005;29:755-759. ECS Stimulation, Centrally and Peripherally, Favors Metabolic Processes that Lead to:: ECS Stimulation, Centrally and Peripherally, Favors Metabolic Processes that Lead to: 36 Weight Gain Lipogenesis Insulin Resistance Dyslipidemia Impaired Glucose Homeostasis Slide37: ECS stimulation centrally and peripherally favors metabolic processes that lead to: Weight gain Lipogenesis: through stimulation of hepatic lipogenic enzymes and increased fatty acid synthesis Altered glucose homeostasis: overactivation of the ECS is associated with increased insulin and glucose levels Dyslipidemia: diet-induced obesity, associated with ECS up-regulation, produces increased levels of LDL, HDL, total cholesterol, and triglycerides Supporting Evidence: Non-Clinical 37 Slide38: In animal models, CB1 blockade reverses or ameliorates endocannabinoid stimulated: Weight gain Impaired glucose homeostasis Insulin resistance Lipogenesis Dyslipidemia Supporting Evidence: Non-Clinical 38 Slide39: STD+vehicle HFD+vehicle HFD+ Rimonabant (10 mg/kg) * * *Pandlt;0.01 vs other groups CB1 Blockade Ameliorates the Glycemic Abnormalities Associated with Diet-induced Obesity in Mice Ravinet Trillou C, et al. Am J Physiol Regul Physiol. 2003;284:R345-353. 39 You do not have the permission to view this presentation. In order to view it, please contact the author of the presentation.
ECS 1 Foundations of Endocannabinoid Biology Peppar Download Post to : URL : Related Presentations : Share Add to Flag Embed Email Send to Blogs and Networks Add to Channel Uploaded from authorPOINT Insert YouTube videos in PowerPont slides with aS Desktop Copy embed code: (To copy code, click on the text box) Embed: URL: Thumbnail: WordPress Embed Customize Embed The presentation is successfully added In Your Favorites. Views: 1005 Category: Science & Tech.. License: All Rights Reserved Like it (1) Dislike it (0) Added: August 06, 2007 This Presentation is Public Favorites: 1 Presentation Description No description available. Comments Posting comment... By: hbalzac4 (15 month(s) ago) very well done if you will allow me to download I can use in my work/research Saving..... Post Reply Close Saving..... Edit Comment Close By: psychiatristme (16 month(s) ago) i feel sir this presentation is very comprehensive & extremly helpful for students of various fields,includin me a psychiatrist,i will be highly obliged if you allow me to download this presentation so that i will be able to prepare presentations with such fine competency.thank you Saving..... Post Reply Close Saving..... Edit Comment Close By: shankark (25 month(s) ago) Hello : I find his presentation very helpful. Is there is any way, I can get the ppt. slides? Thanks Saving..... Post Reply Close Saving..... Edit Comment Close By: feky (25 month(s) ago) very helpfull Saving..... Post Reply Close Saving..... Edit Comment Close Premium member Presentation Transcript The Endocannabinoid System and Its Role in Obesity and Cardiovascular Disease: The Endocannabinoid System and Its Role in Obesity and Cardiovascular Disease Discovery Timeline of the Endocannabinoid System: Discovery Timeline of the Endocannabinoid System 2600 BC – Chinese emperor Huang Ti advised taking Cannabis for relief of cramps, and rheumatic and menstrual pain 1839 – Publication by British physician O'Shaughnessy publicizes therapeutic potential of Cannabis to Western world 1964 – Isolation of 9-THC, the active constituent of Cannabis sativa 1991 – Cloning of the human CB1 receptor 1988 – First identification of cannabinoid receptors in rat brain 1990 – Cloning of the first G protein- coupled cannabinoid receptor, CB1 in rat Discovery Timeline (cont’d): 1992 – Discovery of anandamide, the first endogenous cannabinoid 1993 – Cloning of the CB2 receptor 1994 – Characterization of the first CB1 receptor inhibitor, SR141716 (rimonabant) 1995 – 'Endocannabinoid' coined by Di Marzo 1995 – Isolation of a second endocannabinoid, 2-AG 2000s – Wide tissue distribution of CB1 receptors Characterization of peripheral ECS Role of ECS in human obesity and obesity-related metabolic disorders Discovery Timeline (cont’d) Foundations of Endocannabinoid Biology: Foundations of Endocannabinoid Biology Receptors Ligands Pathways of synthesis and degradation Signaling Introduction to physiological effects 4 Slide5: I. The Endocannabinoid System (ECS): Receptors CB1 are located in several areas of the brain and in a variety of tissues such as adipose tissue, liver, muscle, the gastrointestinal tract, pancreas, gonad and sensory neurons CB2 receptors are located in immune cells such as T-cells T-cells B-cells Monocytes Spleen Tonsils 5 Howlett AC et al., Pharmacol Rev. 2002, 54: 161-202. Slide6: II. The ECS: Ligands that Bind to Cannabinoid Type 1 (CB1) Receptors Plant-derived Endogenously produced 3 types: endogenously produced, plant-derived, and synthetic Endogenous ligands (endocannabinoids) synthesized from membrane-derived phospholipids As lipophilic compounds, endocannabinoids are not stored within the synaptic vesicle cytoplasm, but can diffuse freely across cell membranes Di Marzo V, et al. Nat Neurosci. 2005;8:585-589. 6 D9-Tetrahydrocannabinol Slide7: III. The ECS: Pathways of Ligand Synthesis and Degradation Endocannabinoids are produced on demand from phospholipid-derived precursors in the cell membrane Anandamide and 2-AG act as retrograde messengers Fatty Acid Amide Hydrolase rapidly degrades these ligands 7 Freund TF et al. Physiol Rev. 2003, 83: 1017-1066. Classical neurotransmitter: Acetylcholine (Ach): Classical neurotransmitter: Acetylcholine (Ach) IV. The ECS Signaling: Contrast Between Classical Neurotransmission and the ECS 8 Di Marzo V, et al. Nat Rev Drug Discov. 2004;3:771-784. Slide9: Di Marzo V, et al. Nat Rev Drug Discov. 2004;3:771-784. V. The ECS: Cellular Effects of CB1 Receptor Activation 9 Gaps in Current Understanding of the Endocannabinoid System: Gaps in Current Understanding of the Endocannabinoid System How are endocannabinoids taken back up into cells? What is the relevance of circulating endocannabinoid levels? If they are produced on demand and degraded immediately, how can circulating levels be measured? How do endocannabinoids act in peripheral tissues? Where do they come from? Are peripheral effects dependent on neuronal innervation? 10 Gaps in Current Understanding of the Endocannabinoid System: Gaps in Current Understanding of the Endocannabinoid System What are the relative contributions of central and peripheral endocannabinoid system activities to the overall clinical impact of CB1 blockade? To what extent are metabolic effects of CB1 blockade dependent on concomitant reductions in food intake and body weight, and to what degree are they attributable to direct pharmacological effects? 11 Some CB1 Receptor Antagonists Currently in Development: Some CB1 Receptor Antagonists Currently in Development Phase 1 Solvay Pharmaceuticals, Bristol-Myers Squibb SLV 319 Phase 2,3 Merck L-000899055 (MK0364) Phase 1 Pfizer CP-945598 Under FDA review Sanofi-Aventis Rimonabant (SR141716) andamp; SR141778 Stage of Development Manufacturer Drug 12 The ECS Modulates:: The ECS Modulates: 13 Energy Balance Feeding Behavior Hepatic Lipogenesis Glucose Homeostasis Sites of Action for CB1 Modulation of Energy Homeostasis: Energy Balance Feeding Behavior Gastric emptying GI motility Hepatic glucose output Lipogenesis Hepatic Lipogenesis Adipose Tissue Metabolism Glucose Homeostasis Glucose uptake Glucose, lipid oxidation Lipolysis Lipogenesis Ghrelin, PYY Limbic forebrain Motivation for palatable food Sites of Action for CB1 Modulation of Energy Homeostasis 14 Slide15: Energy balance EC blockade reduces food intake and body weight CB1 deletion is associated with a lean phenotype and resistance to diet-induced obesity Feeding behavior Stimulation of CB1 receptors increases food intake whereas CB1 blockade reduces food intake Supporting Evidence: 15 Wiley JL et al. Br J Pharmacol. 2005;145:293-300. Cota D et al. J Clin Invest. 2003;112(3):423–431. Hao S et al. Eur J Pharmacology. 2000; 392:147-156. Endocannabinoids Stimulate Food Intake in Mice: Endocannabinoids Stimulate Food Intake in Mice Hao S et al. Eur J Pharmacology. 2000; 392:147-156. Anandamide 0.001 mg/kg Vehicle *Pandlt;0.05; **Pandlt;0.01 vs vehicle 1 3 5 7 Day Food intake (grams/day) 7 6 5 4 3 * * * ** 16 CB1 Blockade Produces a Dose-Related Reduction in Food Intake in Mice: CB1 Blockade Produces a Dose-Related Reduction in Food Intake in Mice Wiley JL et al. Br J Pharmacol. 2005;145:293-300. 2.0 1.5 1.0 0.5 0.0 0.0 0.3 1.0 3.0 10.0 Rimonabant Dose (mg/kg-1) Food intake (g) 17 Slide18: CB1 Knockout (CB1-/- ) Mice Sustain a Reduced Body Weight vs. Pair-fed Wildtype Mice Cota D et al. J Clin Invest. 2003;112(3):423–431. Pair-feeding showed that activation of metabolic processes other than reduced food intake contribute to the lean phenotype in CB1 knockout mice 20 21 22 15 20 25 30 35 40 45 Age (week) Body weight (g) 18 CB1+/+ WILDTYPE CB1+/+ WILDTYPE CB1-/- KNOCKOUT Pair-fed Slide19: Hepatic lipogenesis High fat diet and CB1 activation increase hepatic SREBP-1c and the associated lipogenic enzymes FAS and ACC CB1 blockade attenuates these changes and the fatty liver that occurs as a consequence CB1 knockout animals are resistant to increased hepatic SREBP-1c and hepatic FAS and ACC expression as well as the resulting fatty liver Supporting Evidence: 19 Osei-Hyiaman D, et al. J. Clin. Invest. 2005;115:1298-1305. The ECS is Overactivated in Liver of Diet-Induced Obese Animals, Leading to Fatty Liver in Wildtype but not CB1 Knockout Animals: The ECS is Overactivated in Liver of Diet-Induced Obese Animals, Leading to Fatty Liver in Wildtype but not CB1 Knockout Animals Osei-Hyiaman D, et al. J. Clin. Invest.2005;115:1298-1305. Hepatic anandamide (AEA) levels are elevated in diet-induced obesity, more in wildtype vs CB1 knockout animals AEA (fmol/mg tissue) 20 Slide21: A. CB1 activation increases while CB1 blockade reduces expression of lipogenic enzymes Osei-Hyiaman D, et al. J. Clin. Invest. 2005;115:1298-1305. 21 Vehicle CB1 CB1 CB1 agonist + agonist blockade blockade 300 200 100 0 200 100 0 200 100 0 * * SREBP-1c mRNA (% of control) FAS mRNA (% of control) ACC1 mRNA (% of control) *Pandlt;0.05 vs Vehicle Fatty acid synthase Acetyl CoA carboxylase Hepatic Lipogenesis is Modulated by CB1 Activation Slide22: Osei-Hyiaman D, et al. J. Clin. Invest. 2005;115:1298-1305. 22 Rate of fatty acid synthesis (mmol/g/h) 80 40 0 Normal diet High fat diet Vehicle Rimonabant Vehicle Rimonabant * B. De novo hepatic fatty acid synthesis is increased by high-fat diet through CB1 receptor activation and is attenuated by CB1 blockade Hepatic Lipogenesis is Modulated by CB1 Activation *P andlt; 0.01 vs. vehicle Slide23: Glucose homeostasis After high fat feeding, CB1 knockout animals have a lower glucose response to insulin injection than wild-type animals Administration of CB1 blockade for 7 days to obese animals leads to increased insulin-stimulated glucose uptake by isolated soleus muscle Supporting Evidence: 23 Ravinet Trillou C et al. Int J Obes. 2004;28:640–648. Liu YL et al. Int J Obes Relat Metab Disord. 2005;29:183-187. Slide24: CB1 Knockout Mice Are Leaner and Resistant to Diet-Induced Obesity and Its Metabolic Consequences Ravinet Trillou C et al. Int J Obes. 2004;28:640–648. **Pandlt;0.01 vs CB+/+ on standard diet Standard High fat Glucose response to intraperitoneal (IP) insulin 200 150 100 50 0 ** Glucose AUC 0-180 min Glycemia AUC from 0-180 min after i.p. insulin (0.6 U/kg) injection CB1-/- Knockout CB1+/+ Wildtype 24 Slide25: Effect of 7-day treatment with Rimonabant (10 mg/kg; i.p. once daily) on glucose uptake (formation of 2-deoxy[3H]-glucose-6-phosphate) in isolated soleus muscle in the presence of 10 nM insulin CB1 Receptor Blockade Stimulates Muscle Glucose Uptake in Obese Rats Muscle glucose uptake Liu YL et al. Int J Obes Relat Metab Disord. 2005;29:183-187. 25 Slide26: Adipose tissue metabolism EC stimulation with CB1 agonist increases adipose tissue LPL expression while CB1 blockade inhibits this effect CB1 stimulation reduces while blockade increases adiponectin synthesis CB1 blockade reverses the histological changes in adipose tissue produced by diet-induced obesity EC stimulation reduces the expression of AMP kinase in visceral fat Supporting Evidence: 26 Cota D et al. J Clin Invest. 2003;112:423. Matias I, et al. XV ICRS Symposium June 24-27, 2005; Clearwater, Fla. Jbilo O, et al. FASEB J. 2005;19:1567-1569. Slide27: The Peripheral ECS in Adipose Tissue Cota D et al. J Clin Invest. 2003;112:423. * Pandlt;0.05 vs vehicle control † Pandlt;0.05 vs CB1 agonist Rimonabant inhibits the increase in lipoprotein lipase expression produced by CB1 stimulation 300 200 100 0 LPL activity (% of control) - + + - - - + + CB1 agonist Rimonabant * † † 27 Slide28: The Peripheral ECS in Adipose Tissue Matias I, et al. XV ICRS Symposium June 24-27, 2005; Clearwater, Fla. CB1 stimulation significantly reduced, while rimonabant increased, adiponectin. 28 Relative Adiponectin Expression on day 12 ** ** Control CB1 CB1 stimulation blockade (HU-210) (Rimonabant) **Pandlt;0.01 vs Control Slide29: The Peripheral ECS in Adipose Tissue Adipose tissue of obese mice fed a high fat diet (HFD) plus rimonabant resembles that of lean mice fed a standard diet (STD) Jbilo O, et al. FASEB J. 2005;19:1567-1569. 29 The ECS is Overactivated in:: The ECS is Overactivated in: 30 Animal models of genetic obesity Animal models of diet-induced obesity Human obesity Supporting Evidence: Supporting Evidence Animal models of genetic obesity Increased endocannabinoid levels in several genetic obesity models Increased endocannabinoid levels and CB1 receptor expression in diet-induced obesity 31 Bensaid M, et al. Mol Pharm. 2003;63:908-914. The ECS Is Overactivated in Adipose Tissue of Genetically Obese Rats: The ECS Is Overactivated in Adipose Tissue of Genetically Obese Rats Bensaid M, et al. Mol Pharm. 2003;63:908-914. 32 CB1 Receptor mRNA Level (Arbitrary Units) 0 1 2 3 4 5 Lean Obese Rats U 3T3 F442A Cells 0 1 2 3 4 5 0 1 2 3 4 5 D Up-regulation of CB1 receptor mRNA In white adipose tissue of obese (fa/fa) vs. lean Zucker rats) In mouse 3T3 F442A adipocyte cells Slide33: 10 8 6 4 2 0 16 14 12 10 8 5 4 18.5-24.9 25.0-29.9 andgt;30.0 BMI (kg/m2) 18.5-24.9 25.0-29.9 andgt;30.0 Caucasians African Americans Pandlt;0.05* Pandlt;0.01* Pandlt;0.05* BMI (kg/m2) Percent of subjects with FAAH 385 A/A genotype by BMI category % of subjects with FAAH 385 A/A * vs normal BMI A Mutation in the Enzyme That Degrades Endocannabinoids is Associated with Increased BMI Sipe JC et al. Int J Obes Relat Metab Disord.2005;29:755-759. 33 Slide34: Engeli S, et al. Diabetes 2005; 54:2838–2843. * Pandlt;0.05 vs lean women The ECS is Upregulated in Human Obesity 34 Supporting Evidence: Supporting Evidence The endocannabinoid system is up-regulated in human obesity Missense mutation in FAAH, the primary enzyme involved in endocannabinoid degradation, is associated with increased BMI Circulating endocannabinoid levels are increased, while FAAH is reduced in obese women FAAH expression in adipose tissue is reduced in obese vs. lean women 35 Sipe JC et al. Int J Obes Relat Metab Disord.2005;29:755-759. ECS Stimulation, Centrally and Peripherally, Favors Metabolic Processes that Lead to:: ECS Stimulation, Centrally and Peripherally, Favors Metabolic Processes that Lead to: 36 Weight Gain Lipogenesis Insulin Resistance Dyslipidemia Impaired Glucose Homeostasis Slide37: ECS stimulation centrally and peripherally favors metabolic processes that lead to: Weight gain Lipogenesis: through stimulation of hepatic lipogenic enzymes and increased fatty acid synthesis Altered glucose homeostasis: overactivation of the ECS is associated with increased insulin and glucose levels Dyslipidemia: diet-induced obesity, associated with ECS up-regulation, produces increased levels of LDL, HDL, total cholesterol, and triglycerides Supporting Evidence: Non-Clinical 37 Slide38: In animal models, CB1 blockade reverses or ameliorates endocannabinoid stimulated: Weight gain Impaired glucose homeostasis Insulin resistance Lipogenesis Dyslipidemia Supporting Evidence: Non-Clinical 38 Slide39: STD+vehicle HFD+vehicle HFD+ Rimonabant (10 mg/kg) * * *Pandlt;0.01 vs other groups CB1 Blockade Ameliorates the Glycemic Abnormalities Associated with Diet-induced Obesity in Mice Ravinet Trillou C, et al. Am J Physiol Regul Physiol. 2003;284:R345-353. 39