Overview Personal Genomics Nutrition and Cancer A N Tony

Overview: Personal Genomics Nutrition and Cancer A.-N. Tony Kong, Ph.D. Center for Cancer Prevention Director, Graduate Program In Pharmaceutical Science Ernest Mario School of Pharmacy Rutgers, The State University of New Jersey The 2008 Research Conference on Food, Nutrition, Physical Activity & Cancer November 6 & 7, 2008, Capital Hilton Hotel, Washington DC SNPs facts More than 3 billion base pairs in 23 pairs of chromosomes exist in a human cell. Two single nucleotide polymorphisms (SNPs) occur in about 2,200 nucleotides (0.09%). Approximately 10 million SNPs estimated to occur commonly in our human genome The number of tag SNPs that contain most of the information about the patterns of genetic variation is estimated to be about 300,000 to 600,000, which are far fewer than the 10 million common SNPs. Genome-Wide Associations (GWA) Karyogram http://www.hapmap.org/karyogram/gwas.html Genome-Wide Associations (GWA) Karyogram Genome-Wide Associations (GWA) Karyogram Nutrigenomics Personalized nutrigenomics - the use of molecular analysis to better manage an individual’s respond to nutritional factors so that optimal health outcomes can be achieved by choosing the best approaches (PK & PD) suited to their genetic and environmental/nutrition profile. The response to dietary food factors is determined in part by the effective concentrations of the bioactive food components reaching the target tissues and the threshold amount requires to elicit a response. Hence this threshold response to food factors will vary from individual to individual depending on their genetic make ups. 6 Intestine Not Phytochemicals absorbed Absorption Efflux Epithelium Colon Gut microflora -Hydrolysis -Fermentation Absorption Epithelium Fecal excretion Metabolism -Enteric hydrolysis -Phase II Metabolism -Phase II Portal circulation Liver Metabolism -Phase I -Phase II Biliary excretion - Phase III : Efflux transport Systematic circulation Metabolism & PD Receptor interactions Signaling interactions Tissues Renal excretion -Efflux transport Sources of variation in phytochemicals metabolism and disposition Adopted from Johanna W. Lampe and Jyh-Lurn Chang, Semin Cancer Biol. 2007 Kidney Polymorphism of Phase II GSTs GSTM1 and GSTT1, deletions resulting in no enzyme activity in about 53% and 20% of Caucasian populations (CEBP 2001) GSTP1, a single nucleotide polymorphism on exon 5 resulting an amino acid substitution (Ile105Val) with reduce activity towards Sulforaphane; with 38-49% (+/-) and 7-12% (-/-) (Pharmacogenetics 2001) GSTA1, a haplotype with SNP in the SP1-binding element (C-69T), reduces expression in ~15% of population (Pharmacogenetics 2001) 8 GST Null Genotypes, Crucifers & Colon Cancer Lin et al. reported GSTM1 null genotype (from Southern California) with high broccoli intake had lower prevalence of colorectal adenomas (CEBP 1998) Seow et al. showed that only GSTM1,T1 null genotypes (not single; Singapore), associated with lower risk of colon cancer with high cruciferous vegetables (Carcinogenesis 2002) However, other studies including Tijhuis et al. reported GSTP1 (A331G) and GSTA1 (C-69T) (low expression; from Netherlands) had increase risk of colorectal adenomas with higher cruciferous 9 vegetables intake (2005). GST Genotypes, GST induction and Pharmacokinetics (PK) of ITCs Dr. Lampe et al. showed that in healthy subjects receiving 6-day brassica vegetables, a 26% increased vs 18% increased of GST-α in GSTM1-null over GTSM1+ women (CEBP, 2000) Dr. Zeisel et al. as well as others reported there were no substantial differences in 24-hr urinary ITC levels among all 4 GST genotypes, either individually or in combination following a known dose of broccoli diet (J. Nut. 2006). 10 GSTp1/p2 -/- Mice have increased skin tumorigenesis Hendersen et al. showed that GSTp1/p2 null mice had increased skin papillomas initiated by DMBA and promoted by TPA as compared to the wild-type mice (PNAS 1998). Suggesting that GST enzyme protects against carcinogenesis It is unclear why there is a difference (if any) between mutations of GST in human versus in the mice (many other genes mutations in human?) Nrf2 KO mice models in carcinogenesis and cancer 11 chemoprevention by dietary phytochemicals Healthy Compounds Isolated from Fruits, Vegetables and Tea Tea polyphenols – Green & black Tea HO OH OH O O HO O OH OH OH Epigallocatechin-3-gallate OH Curcumin – Turmeric curry O O O O Curcumin O O Isothiocyanates – Brussels Sprout, broccoli H3C N S O Sulforaphane C S Cancer, Cardiovascular, CNS, Inflammatory and Metabolic Dis. Concept of Chemoprevention Chemopreventive Compounds Blocking Agents Suppressing Agents Normal Cells Initiation Mutant Cells Promotion Benign Tumors Progression Malignant Tumors Carcinogenesis Lee Wattenberg, 1971 Phytochemicals-induced Chemical Stress – ITC, CUR,EGCG Chem/Oxidative Stress GSH / Protein Thiols Pre-Initiated Cells Normal Cells Survival Response Ca++ Ras Raf MEK1/2 ERK ARE: Defense Genes - GST, QR, MT, HO Apoptotic Response Mitochond Bcl-2 PKC MEKK1-3 TAK1 NFκB IκB . MEK3/6 Caspases 3 (CPP32), 6, 7 MEK4/7 JNK p38 AP-1, NFκB Survival Genes: c-Jun, c-Fos, cdks Apoptosis Concept of Chemoprevention Chemopreventive Compounds Blocking Agents Suppressing Agents Normal Cells Initiation Mutant Cells Promotion Benign Tumors Progression Malignant Tumors Carcinogenesis Lee Wattenberg, 1971 Blocking Agents • Blocking activation of potential carcinogens (P450) Induce Phase II Detoxifying and Cellular Defense Enzymes – more critical? • Isothiocyanates – water cress, broccoli, cauliflower CH3 S O Sulforaphane N C S Induction of ARE-mediated Reporter Gene By Various Isothiocyanates in HepG2 Cells Tom Rushmore & Cecil Pickett Pharm. Res. 13: 1043, 1996 Induction of Luciferase by Green Tea Compounds 16 14 12 10 8 6 4 2 0 Catechin 0 uM Fo ld EC 25 uM ECG 50 uM 100 uM EGC 250 uM EGCG 500 uM GTP Arch. Pharmacal. Res. 605-612, 2000 MAPK Cascade in Phase 2 Enzymes Induction Phase 2 Enzyme Inducers BHA, tBHQ, PEITC, SUL Ras MAPKKK Raf MEKK1/ASK1 TAK1 MAPKK MAPK Aim 1 MAPK Pathways MEK1/2 ERK SEK (MKK4/7) MKK3/6 p38 Aim 3 Phosphorylation Kong, 1995 + JNK + Nrf2/Maf - Aim 2 Transcription ARE-Mediated Phase 2 Gene NQO1 Effects of PEITC (water cress) on MAPK (JNK) PEITC Activates JNK1 Activity in HeLa and HT1080 Sarcoma Cells HeLa Cells PEITC (µΜ) 0 1 µΜ) 5 10 30 50 100 300 500 GST-c-Jun HT 1080 Cells PEITC (µΜ) µΜ) 0 10 30 50 100 NaCl GST-c-Jun HeLa Cells Time (min) 0 15 30 60 90 120 180 240 360 GST-c-Jun Cancer Res 56: 2954, 1996 Sulforaphane Activates ERK1/2 MAPK in vivo rat liver Figure 3 E – Sulforaphane Treated Rat Livers Phos phor ylati on of MAPKs after 50 µ mol S UL/rat Time: 0 1 2 4 8 12 16 24 36 48 (h) P hospho-p44 P hospho-p42 Fold: 1.0 2.2 3.6 3.0 3.0 2.7 1.0 1.9 2.8 3.4 Time : 0 Fold: 1.0 1 2 4 8 12 16 24 36 48 (h) P hospho-p54 P hospho-p46 0.9 1.0 0.9 0.9 1.0 1.1 1.1 1.2 0.9 Time : 0 Fold: 1.0 1 2 4 8 12 16 24 36 48 (h) P hospho-p38 1.0 1.0 1.0 1.0 1.0 1.0 1.0 0.9 0.9 Time: 0 1 2 4 8 12 16 24 36 48 (h) Actin Fold: 1.0 1.0 1.0 0.9 0.9 1.0 1.0 1.1 1.0 0.9 Hu et al., JPET, July 2004 MAPK Cascade in Phase 2 Enzymes Induction Phase 2 Enzyme Inducers BHA, tBHQ, PEITC, SUL Ras MAPKKK Raf MEKK1/ASK1 TAK1 MAPKK MAPK Aim 1 MAPK Pathways MEK1/2 ERK SEK (MKK4/7) MKK3/6 p38 Aim 3 Phosphorylation Kong, 1995 + JNK + Nrf2/Maf - Aim 2 Transcription ARE-Mediated Phase 2 Gene NQO1 Regulation of Nrf2 signaling by Phytochemicals Kinas e Dietary Chemopreventive Compounds Transform Translocation Activate Antagonize/inhibit Yu et al., Curr Cancer Drug Targets. 2007 Aug B-ZIP Transcription Factor Nuclear Factor-Erythroid 2 (NF-E2)Related Factor 2 (Nrf2) Pol-II Keap1 Nrf2-wt 1 Neh-2 Neh-4 S S Neh-5 S Neh-6 SS CNC DNA Zipper S Neh-3 589 CBP Small Maf Negative regulation Transcription Activation Neh-1 DNA binding & heterodmerization YW Kan, PNAS, 1994 Nrf2 was cloned using AP-1 site probe to screen a lambda gt11 cDNA expression library from K562 cells PEITC Releases Nrf2-GFP from Keap1-DsRED EGFP-Nrf2 + DsRed-Keap1 DAPI Keap1 Nrf2 Merged EGFP-Nrf2 + DsRed-Keap1+ PEITC DAPI Keap1 Nrf2 Merged Molecular Cancer Ther. Aug 2006 ERK Releases Nrf2-GFP from Keap1-DsRED MEK1+ERK2+EGFP-Nrf2+DsRed-Keap1 DAPI Keap1 Nrf2 p-ERK DAPI Keap1 Nrf2 DAPI Keap1 pERK DAPI Nrf2 p-ERK DAPI Keap1 Nrf2 p-ERK Molecular Cancer Ther. Aug 2006 Phosphorylation of Nrf2 by MAPK Nrf2 c-Jun ATF2 MBP MBP + Nrf2 + - + c-Jun JNK1 - + Nrf2 - + Nrf2 + ATF2 p38δ δ Erk2 Phosphorylation of Nrf2 by ERK2/MEK1 increases Nrf2 binding to small Maf proteins Erk2: MEK1: Nrf2: p-Erk1/2 Nrf2 sMafs Input + + + + 2 + + 0.8 Erk2: DNEE-MEK1: GST-Nrf2 p-T/S-P GST-Nrf2 is phosphorylated by Erk2 ’n -p ST rf2 rf2 G N N Nrf2 Nrf2 IP: GFP sMafs sMafs Co-transfection of Nrf2 with Erk2/MEK1 and MafG increases Nrf2 binding to small Maf proteins in the cells Input sMafs GST pull down + + + In vitro phosphorylation increases GST-Nrf2 binding to small Maf proteins The localization of Nrf2 is regulated by a balance between multiple NLS/NES Neh2 Neh4 Neh5 NESTA 176 Neh6 CNC + + bNLS Neh1 LLLLLL Neh3 NESzip b3 b4 b5 b6 L-S-I-P-E-L-Q-C-L-N-I186 C183: L – Q – C (IAB) – L – N – I – E y5 JBC Aug 2005 The NESTA of Nrf2 can sense redox signal and translocate into nucleus FRET analysis indicates NESTA is not binding with Keap1 NESTA is translocated into nucleus upon oxidative stimuli EGFP-NESTA CFP-NESTA + YFP-Keap1 +DMSO 255 +DEM +H2O2 +tBHQ CFP-Nrf2NT + YFP-Keap1 0 +GSH +Sul +GSH & Sul +Sul YFP Channel CFP Channel FRET Channel EGFP-NESTA EGFP-NESTA C183A JBC Sep 2006 Redox Regulation of Nrf2: In vitro signaling circuitry Dietary phytochemicals activate kinases, increase Nrf2 protein level and induce detoxifying enzymes MAPKs phosphorylate Nrf2 in vitro and in vivo. Phosphorylation of Nrf2 increases its binding to small Maf proteins and potentiates Nrf2/ARE signaling. Small Maf proteins compete with Crm1 to sequester Nrf2 in nucleus, thus stabilize Nrf2 protein. The localization of Nrf2 is regulated by multiple NLS/NES. Nrf2 itself could sense redox signal and shuttle between cytoplasm and nucleus Keap1 dimer BTB BTB SH SH Unstimulated SH SH Nuclear Translocation DLG Current Nrf2 Model ETGE Ub Ub Ub Ub kNrf2 Total Nrf2 fNrf2 Proteasomal Degradation Protein Translation Keap1 dimer Oxidative Stress Stimulated by Oxid. Stress / ITCs, CUR/DBM Mol. Carcinogen. 2008 BTB S S BTB S S Nuclear Translocation Kinases kNrf2 Total Nrf2 fNrf2 ETGE G DL Proteasomal Degradation Protein Translation Nrf2 binds to p160 SRC and modulate Nrf2/ARE transcription JBMB, 2006 SH P Chemical Signal Kinases Keap1 Nrf2 SH Maf ARE 300 P/P CB CoA CARM AN 1 OC SRCCo P/C AF SAM Methyl Acetyl Ac Co A Phase II Gene Expression POL II SM RT 1 TFIIB TBP Complex Nrf-2 knock-out Mouse Vehicle Preventive Compounds Wild Type wt/V wt/Rx Nrf-2 -/- ko/V ko/Rx Affy 45 K Probes Chips Gene expression profiling using Nrf2 (-/-) and wild type C57BL/6J mice VEHICLE TREATMENT VEHICLE TREATMENT (+/+) (+/+) (-/-) (-/-) 45 K Probes Affinity Chips Nrf2-dependent genes Genes differentially regulated over 2-fold in Nrf2(+/+) mice Genes differentially regulated over 2-fold in Nrf2(-/-) mice SFN 3 h Nrf2 WT C57B6 vs Nrf2 KO - liver ARE-Mediated Detoxifying Genes 1448239_at 12.24 1424296_at 5.74 1455959_s_at 4.85 1421041_s_at 4.08 1421040_a_at 2.80 1419435_at 2.67 1451612_at 2.66 1423869_s_at 2.44 MAPK Pathways 1419248_at 1418726_a_at 1425575_at 1456927_at 1450036_at 1427083_a_at 1416272_at 1419055_a_at 1452697_at 1452318_a_at 1442767_s_at 1416681_at 1450161_at 1454690_at 1434633_at 1420444_at heme oxygenase (decycling) 1 glutamate-cysteine ligase, catalytic subunit glutamate-cysteine ligase, catalytic subunit glutathione S-transferase, alpha 2 (Yc2) glutathione S-transferase, alpha 2 (Yc2) aldehyde oxidase 1 metallothionein 1 thioredoxin reductase 3 42.47 27.14 24.92 12.70 10.51 7.63 2.40 9.82 6.19 10.35 7.01 3.61 4.40 3.54 4.25 3.84 regulator of G-protein signaling 2 troponin T2, cardiac Eph receptor A3 microtubule associated serine/threonine kinase 2 serum/glucocorticoid regulated kinase 3 mitogen-activated protein kinase kinase kinase kinase 5 mitogen-activated protein kinase kinase 1 interacting protein 1 protein tyrosine phosphatase, non-receptor type 21 CTD (carboxy-terminal domain, RNA polymerase II, polypeptide A) phosphatase, subunit 1 heat shock protein 1A ubiquitin-activating enzyme E1, Chr X ubiquitin protein ligase E3A inhibitor of kappaB kinase gamma inhibitor of kappaB kinase gamma CREB binding protein solute carrier family 22 (organic cation transporter), member 3 EGCG/Curcumin – Small Intestine vs Liver Gene name Homeostasis Curcumin Curcumin EGCG EGCG Liver 3h SIT 3h Liver 3h SIT 3h 4.89 2.12 2.94 heme oxygenase 1 hemopexin thioredoxin reductase 1 thioredoxin interacting protein thioredoxin reductase 2 Transferase glutathione S-transferase alpha 2 (Yc2) glutathione S-transferase mu 1 glutathione S-transferase, mu 3 glutathione S-transferase, alpha 3 glutathione S-transferase, alpha 4 glutathione S-transferase, theta 2 sulfotransferase family 3A, member 1 UDP-glucuronosyltransferase 2 family, member 5 glutamate-cysteine ligase, catalytic subunit methyltransferase-like 1 76.06 7.84 2.35 0.35 4.16 108.00 0.44 9.58 4.42 3.72 2.62 2.49 2.18 2.53 4.82 2.07 2.41 4.28 4.72 2.60 4.52 Nrf2-dependent regulation of Phase II genes by chemopreventive agents in mice liver PEITC SFN 3h 5.7 4.8 4.1 2.0 2.3 2.2 2.0 12h 4.5 4.3 4.4 2.7 4.4 3.1 2.0 2.3 2.2 EGCG 3h 12h 2.6 2.4 2.3 2.7 Curcumin 3h 2.1 2.4 12h 3.2 4.3 2.3 2.7 2.6 Gene Name Phase II Genes GCL, catalytic subunit GCL, catalytic subunit GST, alpha 2 GST, alpha 4 GST, mu 1 GST, mu 3 GST, mu 5 3h 12h 3.6 2.7 2.6 3.6 3.4 Pharm Res., Nov. 2005; Mol. Can. Ther. 2006; Cancer Let, 2006; Life Sci, 2006 Combinations of Phytonutrients • • • Synergistic Effect – CUR + SFN on ARE-activities in HepG2-C8 cells Synergistic Effect of Combination of PEITC and Sulforaphane or Curcumin (CUR) and Sulforaphane (SFN) in the Inhibition of Inflammation. Pharm Res. Oct 2008 Synergistic effect of combination of SFN and EGCG on AP-1 activities in HT-29 cells. Pharm Res. 2008 Nrf2-dependent genes regulated by dietary phytochemicals Detoxification Electron transport Transporters Cell adhesion Cytoskeleton Ubiquitination proteolysis Nrf2/ARE Proliferation Apoptosis Transcription factors Stress response DNA repair Kinase Phosphatase Pharm Res. 2005 Nov; Mol Cancer Ther. 2006 Jan; Pharm Res. 2006 Nov; Cancer Lett. 2006 Nov; J Pharm Sci. 2008 Jan Nrf2 -/- Knock-out Mice Nrf2-deficient mice have an increased susceptibility to dextran sulfate sodium-induced colitis DSS-induced colitis as demonstrated by increased colitis severity after one week oral administration of 1% DSS in Nrf2 -/- mice. Tin Oo Khor, Ph.D. Mou-Tuan Huang, Ph.D. Ki Han Kwon1, Ph.D. Jefferson Y. Chan, Ph.D. Bandaru S. Reddy, Ph.D Cancer Res. Dec 15 2006 Strong Nitrotyrosine immunoreactivity in the colonic crypt cells of DSS-administered C57/B6J (Nrf2 +/+) and Nrf2 (-/-) E F E - Immunoreactivity in Nrf2 (-/-) stronger than in Nrf2 (+/+) mice than in F - C57/B6J (Nrf2 +/+) G H G & H - Undetectable in the untreated control Nrf2 +/+ mice Cancer Res. Dec 15 2006 Nrf2 -/- increased expression of proinflammatory cytokines IL-1β, IL-6, TNF-α & proinflammatory mediators such as iNOS and COX2 BUT decreased Phase 2/antioxidant genes water 1% DSS water 1% DSS Nrf2 (+/+) Nrf2 (-/-) Nrf2 (+/+) Nrf2 (-/-) Nrf2 (+/+) Nrf2 (-/-) Nrf2 (+/+) Nrf2 (-/-) IL-1β β IL-6 TNF-α α iNOS COX-2 HO-1 NQO-1 UGT1A1 GAPDH Cancer Res. Dec 15 2006 Increased Susceptibility of Nrf2 Knockout Mice to Inflammatory Colitis-Associated (AOM-DSS) Colorectal Cancer 100 90 80 Incidence (% ) 70 60 50 40 30 20 10 0 C57BL/6 Nrf2KO (58.6%) tumor (39.6%) prolapsed rectum/ bleeding anus 50 45 40 35 30 25 20 15 10 5 0 WT + DSS/AOM KO + DSS/AOM * n it r o t y r o s in e s c o r e Cancer Prev. Res. Aug 2008 1 2 Sulforaphane suppressed LPS-induced inflammation in mouse peritoneal macrophages through Nrf2-dependent pathway Nrf2 KO macrophages TNFα Nrf2 WT macrophages IL-1β GAPDH LPS(1µg/ml) SFN (µM) + + 40 + 20 + 10 + 5 + + 40 + 20 + 10 + 5 Biochem. Pharmacol. Oct 2008 Nrf2 -/- Knock-out Mice Sulforaphane inhibits skin tumorigenesis in C57BL/6 mice associated with Nrf2 gene 2-Stage Carcinogenesis Model - Mice were treated with 200 nmol DMBA, and one week later, 8 nmol TPA were applied twice a week for 25 weeks. Changjiang Xu, Ph.D. Collaborators: MT Huang, Ph.D. Allan Conney, Ph.D. Cancer Res. Sept 2006 1 .0 Tumor incidence 0 .8 WT W T /S F N N r f2 - /N r f2 - /- /S F N 0 .6 0 .4 0 .2 0 .0 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 T im e ( w e e k s ) Fig 1. Skin tumor incidence. Mice were pretreated with 100 nmol SFN for 14 d, after that, 200 nmol DMBA was applied on the second day, one week later, 8 nmol TPA were applied twice a week for 25 weeks. Cancer Res. Sept 2006 Expression of Nrf2, HO-1, and pro-inflammatory genes in skin tumors Nrf2(+/+) Nrf2 HO-1 Actin c-Myc Bcl-2 Cox-2 VEGF β-catenin Nrf2(-/-) Nrf2(+/+) Nrf2(-/-) Normal samples Tumor samples Xu et al., (2006) Cancer Res. 66:8293-6 TRAMP Mice - Lose of Phase II and Antioxidant Enzymes Expression as Prostate Carcinogenesis Progress Int. J. Cancer 2008 (in press) Restoration of Nrf2, Phase II and Antioxidant Enzymes in the Prostate Tumors in TRAMP Mice Fed with 0.1% γTocopherol Enriched Mixed Tocopherol Diet Int. J. Cancer 2008 (in press) 8 we ek s we ek s we ek s we ek s we ek s 12 16 24 24 Cu rc um Restoration of Phase II UGT1A1 Enzyme in Prostate Tumor of TRAMP mice fed with PEITC, CUR alone and in combination Pharm. Res. Sept 2008 (re pe at ) in (1 PE 6 we IT ek C s) (1 6 we Co ek m s) bin at ion Cu (1 rc 6 um we PE in ek IT (2 s) C 4 we (2 Co 4 we eks m bin ) ek at s) ion (2 4 we ek s) ITCs and APCmin Mice Model • • • SFN inhibited adenomas in APCmin mice dose-dependently (Carcinogenesis 2006) DBM inhibited adenomas and potential synergistic with SFN (Cancer Res 2007) PEITC (0.05% or 500 ppm) inhibited adenomas about the same potency as SFN (Mol. Carcinog. 2008) PEITC and NOT DBM Blocks both initiation and post-Initiation in AOM/DSS Colon Model 90 80 70 60 Tumor incidence 50 40 30 20 10 0 Cont rol PEITC DBM Combinat ion bef ore af t er 2 1.8 Average tumor # 1.6 1.4 1.2 1 0.8 0.6 0.4 0.2 0 Cont rol PEITC DBM Combinat ion bef ore af t er DBM Block carcinogenesis before BUT NOT after PIN Development in TRAMP Mice Model Group 1 (8 weeks) Group 2 (12 weeks) Experiment terminated (24 weeks) 10 weeks Develop PIN 18 weeks Develop invasive adenocarcinoma 24-28 weeks Develop CaP that metastasizes to the lymph nodes and lungs A 35 30 25 20 15 10 5 0 control S1 G1 G2 B 60 50 40 30 20 10 0 control G1 G2 S1 Incidence of palpable tumor (%) Incidence of adenocarcinoma Conclusions • • • • SNPs of Phase 2 (GSTs), antioxidant enzymes and Nrf2 – affect the conc’n levels of dietary components Dietary phytochemicals activate kinases, increase Nrf2 protein level and induce Nrf2-mediated enzymes Nrf2 itself could sense redox signal and shuttle between cytoplasm and nucleus Nrf2 -/- mice are more prone to oxidative stress / inflammatory damages and carcinogenesis Conclusions • • • Phytochemicals CUR, DBM, ITCs PEITC/SFN & γ-Tocopherol inhibited TRAMP, APCmin, and/or AOM/DSS mice carcinogenesis Combinations of some phytochemicals - synergistic Examining the In Vivo Concentration Versus Effect (Response) Relationship of Phytochemicals in human will be critical ACKNOWLEDGEMENTS Current Lab members Wenge Li, Ph.D. Tin Oo Khor, Ph.D. Siwang Yu, Ph.D. Jung Hwan Kim Avantika Gopalkrishnan Wen Lin William Cheung Tien-Yuan Wu Collaborators Bandaru Reddy (Rutgers) Jefferson Chan (UC Irvine) Allan Conney (Rutgers) C. S. Yang (Rutgers) M. T. Huang (Rutgers) Cecil Pickett Xi Zheng (Rutgers) Yue Liu (Rutgers) Xingpei Hao (Rutgers) Hong Li (UMDNJ) Tong Liu (UMDNJ) Alumni Changjiang Xu (CAST) Chi Chen (UM) Rong Hu (NPU) Guoxiang Shen (BMS) Young Sam Keum (UM) Xiaoling Yuan (Yale) Auemduan Prawan (KKU) Supported by NIH Grants CA-073674, CA-094828, CA-118947 AAPS Slide Template [40 pt] For optimum legibility [36 pt] Do not exceed 7 words per line [32 pt] Do not exceed 7 lines per slide Select sans serif fonts (e.g., Arial) Employ Boldface letters KIS (Keep It Simple) 61