Providing the Understanding of Drug Induced Effects on Metabolism Cell Stress Signaling response UPR/ER stress Mito UPR UPR/ER stress Mito UPR Autophagy/Mi tophagy Senescence Apoptosis/ Necroptosis Apoptosis/ Necroptosis Survival THE DRUG INDUCED CELL STRESS RESPONSE

Providing the Understanding of Drug Induced Effects on Metabolism ATP/ADP/AMP. NADH/NAD+ NADPH/NADP+. Membrane potential. ROS/RNS. METABOLITE SHUTTLES Citrate Malate Aspartate Glutamate Pyruvate Acetyl CoA mtUPR. Calcium RETROGRADE SIGNALING FROM MITOCHONDRIA. Where ATP required for transport Where membrane potential/proton gradient drives transport PROTEIN IMPORT/ Nfkappa B, ADHERENCE BCl1, PINK1,MCl1 etc AND RELEASE. AIF,Cytochrome c etc.

Providing the Understanding of Drug Induced Effects on Metabolism Hypoxia Drugs mtDNA depletion ROS ionophores Low Ca+ uptake & ATP down Higher cytosolic Ca+ Low Ca+ efflux calcineurin Activation of NFAT & NFkappaB Ca+ dependent PKC JNK, p38MAPK activation Ca+ induced mitochondrial retrograde response Ryanodine receptor

Providing the Understanding of Drug Induced Effects on Metabolism HSP60 JNK PATHWAY C-JUN PKR eIF2alpha PERK Induces same genes as CHOP1 ATF4 Up reg of CHOP1 Mitochondrial UPR (yellow) ER UPR(blue) APOPTOSIS INHIBIT PROTEIN TRANSLATION AUTOPHAGY p38MAPK HSP90 Up reg of Heme Oxygenase HRI P P P P CELLULAR UNFOLDED PROTEIN RESPONSE eIF2alpha ATG13 P P

Providing the Understanding of Drug Induced Effects on Metabolism P-AMPK S-NITROSYLATION OF JNK1 BECLIN RELEASE FROM BCL2 INACTIVATION OF mTOR AUTOPHAGY/M ITOPHAGY ATG4 HIF1 BECLIN DISSOCIATES FORM BCL2 BNIP3 ??? SSS-NITROSYLATION OF IKK BETA- ROS/RNS Hypoxia H2O2 O2- NO ONOO RETROGRADE SIGNALING INVOLVING ROS AND RNS SOD1 SOD2 Glutathione reductase Peroxyredoxin ATG13

Providing the Understanding of Drug Induced Effects on Metabolism AMPK ACC2 active ACC2 active Acetyl CoA Malonyl CoA PHOSPHOLIPID SYNTHESIS FATTY ACID OXIDATION CONTROL OF LIPID METABOLISM BY AMPK (high ATP) HADHB Citrate,glutamate Dicarboxylic acids Palmitoyl CoA, short & long Chain FAs

Providing the Understanding of Drug Induced Effects on Metabolism AMPK ACC2 inactive ACC2 inactive Acetyl CoA Malonyl CoA PHOSPHOLIPID SYNTHESIS FATTY ACID OXIDATION CONTROL OF LIPID METABOLISM BY AMPK (low ATP) HADHB PI Energy production

Providing the Understanding of Drug Induced Effects on Metabolism PATHWAY OF FATTY ACID SYNTHESIS citrate Mitochondrial Acetyl CoA lipoate elongated saturated fatty acids unsaturated fatty acids palmitate malonyl CoA cytosolic acetyl CoA cytochrome b5 NADH cyt b5 reductase ACC1 ACC2 fatty acid synthase fatty acid synthase ATP citrate lyase ACP Complex I malonyl transferase Pi PiPi PiPi pipi pipi

Providing the Understanding of Drug Induced Effects on Metabolism PATHWAY OFMITOCHONDRIAL FATTY ACID OXIDATION unsaturated F.A. saturated F.A. acyl CoA DH’s acetyl CoA short chain enoyl-CoA hydrolase short chain enoyl-CoA hydrolase LCADVLCAD SCAD SCHA D ETF.Q CPT1 CPT2 DECR ETF HADH B MCAD ACAA2

Providing the Understanding of Drug Induced Effects on Metabolism PEROXISOMAL FATTY ACID METABOLISM H2O2H2O2 phytanic acid pristanic acid C26:0 Ω fatty acids dicarboxylic acids Excretion adipic acid sebacic acid succinate acetyl CoA (chain shortened) ether lipids mitochondria bile salts unsaturated fatty acids taurocholate chenodeoxycholate PBE MFE2 FACL(4) ACCA1 ACOT 4/8 CATALASE ABCD1 ACOX2 ACOX1 BAAT ECH1 PEC1 PHYH/ PAHX CYP 4A IF ALDH 3A2

Providing the Understanding of Drug Induced Effects on Metabolism cholesterol and other sterols D- β hydroxybutyrate acetone +CO 2 acetoacetate Hydroxy- β -methylglutaryl CoA acetoacetyl CoA acetyl CoA D- β hydroxybutyrate dehydrogenase HMG CoA reductase HMG CoA synthase HMG CoA lyase HMG CoA lyase ACAT-1 (mit) ACAT-1 (mit) ACAT-2 cyt KETOGENESIS

Providing the Understanding of Drug Induced Effects on Metabolism AKT SIGNALING PATHWAY AKT active AKT active AKT PI3K PDK1 P P P P FOXO3 inactive FOXO3 inactive P P Bax inactive Bax inactive GSK3 Bad inactiv e Bad inactiv e Casp 9 inactive Casp 9 inactive P P P P Ikk P P Cell survival/proliferation GSK3 inactive P P P P P P mTOR S6 active S6 active S6 P P PI3 MDM2 P P FOXO3 ATP citrate lyase P P P P P53

Providing the Understanding of Drug Induced Effects on Metabolism INHIBITORS OF THE AKT SIGNALING PATHWAY AKT active AKT active AKT PI3K PDK1 P P P P FOXO3 inactive FOXO3 inactive GSK3 Cell survival/proliferation GSK3 inactive P P P P mTOR S6 inactive S6 inactive S6 active S6 active P P PI3 FOXO3 P P WORTMANNIN PIK90 NVP-BEZ235 LY P P RAPAMYCIN NVP-BEZ235 LY LITHIUM

Providing the Understanding of Drug Induced Effects on Metabolism GSK3 active GSK3 active GSK3 PI Glycogen synthase ATP citrate lyase eIF2B Catenins CyclinD1 Transcription of CREB, c-Jun etc. Adhesion and more Protein synthesis Glucose metabolism. Lipid synthesis CDK6 Apoptosis GSK PATHWAY

Providing the Understanding of Drug Induced Effects on Metabolism mTor sK6 4E-BP1 in S6 AKT AMPK GCN PERK eIF2a PROTEIN SYNTHESIS Pi inhibits INHIBITION OF PROTEIN SYNTHESIS AT THE RIBOSOME. Phospho 4E-BP1 & phospho S6 activate translation GSK3 UPR GLUCOSE & FAO METABOLISM eIF2a S6 Pi 4E-BP1 Pi

Providing the Understanding of Drug Induced Effects on Metabolism KEY PATHWAYS THAT CONVERT STRESS TO SURVIVAL OR DEATH P38MAPK active P38MAPK active CHOP ATF2 ERK1/ERK2 (P42/44MAPK) active ERK1/ERK2 (P42/44MAPK) active ASK1 inactive ASK1 inactive AKT active AKT active apoptosis autophagy RAS RAF TRX TRAFs Ox Stress p53 P P P P P P U U JNK C-Jun active C-Jun active P P CHOP P P mTOR PI3K

Providing the Understanding of Drug Induced Effects on Metabolism AUTOPHAGY JNK ERK1/ERK2 PIK3 BECLIN 1 MITOCHONDRIAL DAMAGE= ROS P38 MAPK MTOR CASPASE 8 CONTROL OF AUTOPHAGY BY p38MAPK & ERK1/2 NFkappaB P70s6 K Green=activates Red= inhibits

Providing the Understanding of Drug Induced Effects on Metabolism PGC-1alpha PPARs ERRs NRFs Fatty acid oxidation Glucose metabolismMitochondrial biogenesis AMPK SIRTs mTOR p38MAPK REGULATION OF METABOLISM BY PGC-1ALPHA GLUT4 ERRs = estrogen receptor related proteins