Presentation is loading. Please wait.

Presentation is loading. Please wait.

Tobacco Carcinogen and Toxicant Biomarkers Stephen S. Hecht, Ph.D. Masonic Cancer Center University of Minnesota.

Similar presentations


Presentation on theme: "Tobacco Carcinogen and Toxicant Biomarkers Stephen S. Hecht, Ph.D. Masonic Cancer Center University of Minnesota."— Presentation transcript:

1 Tobacco Carcinogen and Toxicant Biomarkers Stephen S. Hecht, Ph.D. Masonic Cancer Center University of Minnesota

2 Outline of Presentation Tobacco and cancer Background on tobacco carcinogen biomarkers Examples of tobacco carcinogen biomarkers –Total NNAL –Formaldehyde-DNA adducts

3 Cancers Caused by Smoking: IARC Monograph Series Volume 38, 1986 Lung Oral cavity Pharynx Larynx Esophagus Pancreas Bladder Volume 83, 2004, added: Nasal Cavity Stomach Liver Kidney Ureter Cervix Myeloid leukemia Volume 100E, 2009, added: Colorectum Ovary (mucinous)

4 Cancer Deaths Due to Smoking Worldwide: 21% (1,420,000 per year) United States: 33% (185,000 per year) IARC World Cancer Report, 2008

5 Tobacco Use Prevalence, 2008 Adult smokers, U.S.: 46,000,000 (20.6%) Ex-smokers, U.S.: 48,100,000 (21.5%) Smokers, worldwide: 1,300,000,000 Smokeless, worldwide: xxx,000,000 CDC, MMWR, November 13, 2009

6 O. Shafey, M. Ericksen, H. Ross, J. Mackay (2009) The Tobacco Atlas, 3 rd Ed. World Smoking Prevalence: Males

7 O. Shafey, M. Ericksen, H. Ross, J. Mackay (2009) The Tobacco Atlas, 3 rd Ed. World Smoking Prevalence: Females

8 Overall Goal Elucidate mechanisms of tobacco-induced cancer and apply this knowledge to cancer prevention.

9 Conceptual Framework for Understanding Tobacco Carcinogenesis S.S. Hecht, JNCI, 91:1194-1210 (1999), Nature Rev. Cancer 3:733-744 (2003); Cancer: Principles and Practice of Oncology, 8 th Edition, 147-155 (2008)

10 Significantly Mutated Genes in Lung Adenocarcinoma: Based on Sequencing of 623 Genes in 188 Tumors Ding et al, Nature, 455:1069-1075, 2008

11 Other Factors Contributing to Tobacco-Induced Cancer Receptor mediated effects:nicotine, nitrosamines Direct activation of EGFR and COX-2 Down-regulation of FHIT Hyper-methylation of tumor suppressors Tumor promotion and co-carcinogenesis Oxidative damage and inflammation Cilia-toxicity H. Takahashi et al, Cancer Cell 17: 89 (2010); H. Schuller, Nature Rev. Cancer 9: 195-205 (2009); K.A. West et al, J. Clin. Invest. 111: 81-90 (2003); S.A. Belinsky, Carcinogenesis 26: 1481 (2005); D’Agostini et al, Cancer Res 66: 3936-3941 (2006); Jin et al, Carcinogenesis 29: 1614-1622 (2008); Bhutani et al, Cancer Prev. Res. 1: 39-44 (2008)

12 Conceptual Framework for Understanding Tobacco Carcinogenesis S.S. Hecht, JNCI, 91:1194-1210 (1999), Nature Rev. Cancer 3:733-744 (2003); Cancer: Principles and Practice of Oncology, 8 th Edition, 147-155 (2008)

13 IARC Carcinogens in Tobacco Smoke S.S. Hecht, In: DeVita et al, Cancer (2010); IARC Monographs No. 83 (2004); D. Hoffmann and S.S. Hecht, Handbook Exp. Pharmacol. 94:63-102 (1990)

14 Goal Develop and validate tobacco carcinogen and toxicant biomarkers –Urinary metabolites –DNA and protein adducts –Metabolites in blood, saliva, breath, nails, hair Use these biomarkers to identify those smokers susceptible to cancer.

15 Outline of Presentation Tobacco and cancer Background on tobacco carcinogen biomarkers Examples of tobacco carcinogen biomarkers –Total NNAL –Formaldehyde-DNA adducts

16 Definitions Biomarker: A distinctive biological or biologically derived indicator (as a metabolite) of a process, event, or condition (Merriam-Webster’s Collegiate Dictionary) Tobacco carcinogen biomarker: Any quantifiable substance, such as a metabolite, that can be specifically related to human exposure to a given tobacco carcinogen.

17 Tobacco Carcinogen Biomarkers DNA Adducts Protein Adducts –Hemoglobin –Albumin Metabolites –Breath –Saliva –Nails and Hair –Urine –Blood Reviewed in Carcinogenesis 23: 907 and 1979 (2002); Nature Rev. Cancer 3: 733 (2003)

18 Applications of Tobacco Carcinogen Biomarkers Assessing exposure in smokers, smokeless tobacco users, and non-smokers exposed to secondhand smoke Regulation of tobacco products Understanding mechanisms of human carcinogenesis and identifying susceptible individuals Not specifically designed for early detection of cancer, but could have applications in screening

19 Range of recent mean values (nmol/24h unless noted otherwise) Urinary biomarkersSourceSmokersNon-smokers Nicotine equivalentsNicotine70.4-154 µmol/24 hNot Detected Total NNALNNK1.1 - 2.9Not Detected Total NNNNNN0.049 - 0.24Not Detected 1-HOP or PheTPAH0.50 - 1.450.18 - 0.50 MHBMA1,3-Butadiene15.5 - 3220.65 - 7.5 SPMABenzene3.2 - 32.10.17 - 3.14 HPMAAcrolein5,869 - 11,1901,131 - 1,847 HBMACrotonaldehyde1,965 - 26,000242 - 3,200 HEMAEthylene oxide19.1 - 1026.51 - 38.8 CdCadmium2.3 - 12.81.34 - 8.04 8-epi-PGF 2  Oxidative damage1.48 - 2.800.62 - 1.13 PGE-MInflammation54 - 6031.6 - 45.3 Based on 1.3g creatinine per 24h in smokers and 1.5g creatinine per 24h in non-smokers, or 1.5 l urine per 24h. A Panel of Tobacco Carcinogen and Toxicant Biomarkers S.S. Hecht, J-M Yuan, and D. Hatsukami, Chem. Res. Toxicol., 2010

20 Structures of the Urinary Biomarkers

21 Recent data (pmol/g globin; mean ± S.D.) Hemoglobin adductsSourceSmokersNon-smokers CyanoethylvalineAcrylonitrile112 ± 816.5 ± 6.4 CarbamoylethylvalineAcrylamide84.1 ± 41.827.8 ± 7.1 HydroxyethylvalineEthylene oxide132 ± 9221.1 ± 12.7 4-Aminobiphenyl-globin4-Aminobiphenyl0.26 ± 0.006 a 0.067 ± 0.009 a (fmol/µmol dN; mean ± S.D.) Leukocyte DNA adductsSourceSmokersNon-smokers N 6 -hydroxymethyl-dAdoFormaldehyde179 ± 20515.5 ± 33.8 N 2 -ethylidene-dGuoAcetaldehyde1,310 ± 1,720705 ± 438 Mean concentrations OtherSourceSmokersNon-smokers Exhaled COCarbon monoxide17.4 - 34.4 ppm2.6 - 6.5 ppm CarboxyhemoglobinCarbon monoxide3.4 - 7.1 %0.35 - 1.45 % A Panel of Biomarkers

22 Biomarker Validation Analytical –Specificity, sensitivity, accuracy, precision With respect to tobacco –Decreases upon cessation –Dose-response With respect to cancer risk

23 Persistence of Biomarkers Study Smokers provide baseline 24h urine samples. Eight days later, they quit smoking and receive nicotine replacement therapy. They provide 24h urine samples on days 3, 7, 14, 21, 28, 42, and 56 after quitting. Urine samples are analyzed for mercapturic acids (by LC-MS/MS) and other biomarkers. S.G. Carmella, M. Chen, S. Han, A. Briggs, J. Jensen, D. K. Hatsukami, and S. S. Hecht Chem. Res. Toxicol., 22: 734-741 (2009)

24 Metabolism of 1,3-Butadiene to Mercapturic Acids C.L. Sprague and A.A. Elfarra, Chem. Res. Toxicol., 17: 819-826 (2004) a. GSH, GSTs; b.  -glutamyltranspeptidase; c. cysteinylglycine dipeptidase; d. cysteine S-conjugate N-acetyltransferase

25 Mean Urinary MHBMA Reduction Upon Smoking Cessation, N=17

26 Metabolism of Acrolein, Crotonaldehyde, Benzene, and Ethylene Oxide to Mercapturic Acids a. GSH, GSTs; b.  -glutamyltranspeptidase; c. cysteinylglycine dipeptidase; d. cysteine S-conjugate N-acetyltransferase

27 Mean Urinary HPMA Reduction Upon Smoking Cessation, N=17

28 Mean Urinary HBMA Reduction Upon Smoking Cessation, N=17

29 Mean Urinary SPMA Reduction Upon Smoking Cessation, N=17

30 Mean Urinary HEMA Reduction Upon Smoking Cessation, N=17

31 Structures of Urinary Biomarkers S.G. Carmella, et al, Chem. Res. Toxicol., 22: 734-741 (2009)

32 Mean Urinary 1-HOP Reduction Upon Smoking Cessation, N=15

33 Mean Urinary Total NNAL Reduction Upon Smoking Cessation, N=17

34 Outline of Presentation Tobacco and cancer Background on tobacco carcinogen biomarkers Examples of tobacco carcinogen biomarkers –Total NNAL –Formaldehyde-DNA adducts

35 Range of recent mean values (nmol/24h unless noted otherwise) Urinary biomarkersSourceSmokersNon-smokers Nicotine equivalentsNicotine70.4-154 µmol/24 hNot Detected Total NNALNNK1.1 - 2.9Not Detected Total NNNNNN0.049 - 0.24Not Detected 1-HOP or PheTPAH0.50 - 1.450.18 - 0.50 MHBMA1,3-Butadiene15.5 - 3220.65 - 7.5 SPMABenzene3.2 - 32.10.17 - 3.14 HPMAAcrolein5,869 - 11,1901,131 - 1,847 HBMACrotonaldehyde1,965 - 26,000242 - 3,200 HEMAEthylene oxide19.1 - 1026.51 - 38.8 CdCadmium2.3 - 12.81.34 - 8.04 8-epi-PGF 2  Oxidative damage1.48 - 2.800.62 - 1.13 PGE-MInflammation54 - 6031.6 - 45.3 Based on 1.3g creatinine per 24h in smokers and 1.5g creatinine per 24h in non-smokers, or 1.5 l urine per 24h. A Panel of Tobacco Carcinogen and Toxicant Biomarkers S.S. Hecht, J-M Yuan, and D. Hatsukami, Chem. Res. Toxicol., 2010

36 Essential Facts About NNK, A Tobacco-Specific Lung Carcinogen Present in tobacco and tobacco smoke; specific to tobacco products Systemic lung carcinogen in rats, mice, hamsters, and ferrets. Also induces tumors of the pancreas, nasal cavity, and liver in rats Considered to be a cause of lung, oral cavity and pancreatic cancer in people exposed to tobacco products NNK and NNN- Carcinogenic to humans; Group 1 (IARC Volume 89, 2007); reaffirmed (Vol 100E, 2009) S.S. Hecht, Chem. Res. Toxicol. 11:559 (1998); Nature Rev. Cancer 3:733 (2003)

37 Metabolism of NNK by Carbonyl Reduction

38 NNAL Plus NNAL-Glucs (Total NNAL): A Biomarker of NNK Exposure Quantified by GC-TEA or LC-MS/MS High analytical specificity and sensitivity Specific to tobacco product exposure Responsive to dose Measures uptake of a lung carcinogen S.S. Hecht, Carcinogenesis 23 907 (2002); S.G.Carmella et al, CEBP 4: 635 (1995); 12: 1257 (2003); D. Hatsukami et al, Nic. Tob. Res. 8: 169 (2006)

39 GC-TEA Chromatogram of NNAL in a Smoker's Urine

40 Applications of the Total NNAL Biomarker Cessation of smoking or smokeless tobacco Reduction of smoking Carcinogen uptake from new and old tobacco products: –Omni, light and ultra-light cigarettes –Snus and other smokeless products –Ultra low nicotine cigarettes Evaluation of carcinogen dose in various groups –Reducers vs. light smokers –Smokers of differing numbers of cigarettes –Ethnic groups, gender, and teen-age smokers –Smokeless vs. smokers –Duration of smokeless use Carcinogen uptake from secondhand cigarette smoke Relationship to lung cancer D. Hatsukami, J. Jensen, A. Joseph, S. E. Murphy, S.G. Carmella, S.S. Hecht, and co-workers. Cancer Res., JNCI, CEBP, Nic. Tob. Res., 1999-2008

41 Tobacco Harm Reduction: Continuum of Risk Conventional cigarettes Modified tobacco cigarettes Cigarette reduction Cigarette-like delivery devices; Extra-low nicotine cigarettes Smokeless tobacco products Nicotine delivery devices Smoking Cessation Most toxic Least toxic D. Hatsukami et al., Nicotine Tob. Res. 9:S537-S553 2007

42 Applications of the Total NNAL Biomarker Cessation of smoking or smokeless tobacco Reduction of smoking Carcinogen uptake from new and old tobacco products: –Omni, light and ultra-light cigarettes –Snus and other smokeless products –Ultra low nicotine cigarettes Evaluation of carcinogen dose in various groups –Reducers vs. light smokers –Smokers of differing numbers of cigarettes –Ethnic groups, gender, and teen-age smokers –Smokeless vs. smokers –Duration of smokeless use Carcinogen uptake from secondhand cigarette smoke Relationship to lung cancer D. Hatsukami, J. Jensen, A. Joseph, S. E. Murphy, S.G. Carmella, S.S. Hecht, and co-workers. Cancer Res., JNCI, CEBP, Nic. Tob. Res., 1999-2008

43 Non-Smokers’ Exposure to NNK Throughout Life by Measurement of Urinary Total NNAL Exposed GroupType of Exposure Total NNAL (fmol/ml urine) % of Amount in Smokers' Urine a FetusTransplacental 25 ± 29 (amniotic fluid) 1.3 NewbornsTransplacental130 ± 1506.5 Infants (<1 year old)Air83 ± 204.2 Elementary School Children MinneapolisAir56 ± 762.8 MoldovaAir90 ± 774.5 Women Living with Smokers Air50 ± 682.5 Hospital WorkersAir59 ± 283.0 Casino PatronsAir18 ± 150.9 Restaurant and Bar Workers Air33 ± 341.7 a based on 2 pmol/ml total NNAL in smokers S.S. Hecht, Carcinogenesis 23:907 (2002); S.S. Hecht et al, CEBP 15:988 (2006)

44 Total NNAL measurements in secondhand smoke-exposed non- smokers have impact It can only come from secondhand smoke. It represents uptake of a lung carcinogen. It is found in the urine of non-smokers. It is the only lung carcinogen biomarker consistently elevated in exposed non-smokers. These studies should spur clean air legislation in the remaining countries, states and localities.

45 Median Serum Cotinine Levels in Non-Smokers, by Age Group: 1988-2002 CDC NHANES Study; MMWR 55: 1130 (2006)

46 Regulation of Indoor Smoking and Tobacco Control Regulation of indoor smoking –Reduces cues for smoking –Reduces amount smoked –Can change social norms Regulation of indoor smoking, along with counter-advertising and taxation, are the most effective methods in tobacco control.

47 Applications of the Total NNAL Biomarker Cessation of smoking or smokeless tobacco Reduction of smoking Carcinogen uptake from new and old tobacco products: –Omni, light and ultra-light cigarettes –Snus and other smokeless products –Ultra low nicotine cigarettes Evaluation of carcinogen dose in various groups –Reducers vs. light smokers –Smokers of differing numbers of cigarettes –Ethnic groups, gender, and teen-age smokers –Smokeless vs. smokers –Duration of smokeless use Carcinogen uptake from secondhand cigarette smoke Relationship to lung cancer D. Hatsukami, J. Jensen, A. Joseph, S. E. Murphy, S.G. Carmella, S.S. Hecht, and co-workers. Cancer Res., JNCI, CEBP, Nic. Tob. Res., 1999-2008

48 Relationship of Urinary NNAL to Lung Cancer in Two Prospective Cohorts of Cigarette Smokers Collaboration with Professors Mimi Yu and Jian-Min Yuan Two prospective cohorts of Chinese cigarette smokers: Shanghai and Singapore Nested case control study of 246 cases of lung cancer and 245 matched controls Total NNAL and cotinine quantified in stored urine samples collected prior to lung cancer diagnosis

49 Joint Effect of Urinary Total NNAL and Cotinine on Lung Cancer Risk NNAL in tertile Cotinine in tertile 1 st (low)2 nd 3 rd (high) Ca/Co 1 OR (95% CI) 2 Ca/Co 1 OR (95% CI) 2 Ca/Co 1 OR (95% CI) 2 1 st (low)9/471.0023/253.93 (1.54, 10.05)11/105.08 (1.63, 15.89) 2 nd 14/243.01 (1.11, 8.10)31/324.15 (1.70, 10.12)22/264.48 (1.78, 11.31) 3 rd (high)8/103.41 (1.08, 11.25)30/255.58 (2.25, 13.84)93/468.47 (3.69, 19.46) J. Yuan, M. Yu, S.E. Murphy, S. Carmella, S.S. Hecht et al., Cancer Res.,69: 2990 (2009) 1 No. of cases/no. of controls 2 Odds ratios (OR) were adjusted for age, year of interview, year of sample collection, gender and dialect group, study location (Shanghai versus Singapore), number of cigarettes smoked per day, and number of years of smoking; CI, confidence interval.

50 Conclusions of the Shanghai and Singapore Study Total NNAL significantly associated with risk of lung cancer in a dose-dependent manner, after adjustment for smoking history and urinary cotinine. Cotinine was independently associated with lung cancer, consistent with previous data. Smokers in the highest tertiles of urinary total NNAL and cotinine exhibited an 8.5 fold increased risk for lung cancer, relative to those with comparable smoking history, but in the lowest tertiles. J. Yuan et al, Cancer Res. 69: 2990 (2009)

51 Similar Results in the PLCO Study The Prostate, Lung, Colon, and Ovarian Cancer Screening Trial: started 1993 77,468 subjects (25,000 smokers) screened; over 1,000 lung cancer cases diagnosed Questionnaire data and blood samples collected prospectively 100 lung cancer cases and 100 controls without lung cancer selected – all were smokers of > 10 CPD Pre-diagnostic serum analyzed for total NNAL and cotinine T. Church, K. Anderson, M. Geisser, Y. Zhong, C. Le, N. Caporaso, S. Carmella, A. Benoit, S. S. Hecht, CEBP,18: 260 (2009)

52 Total NNAL and Lung Cancer Total NNAL is a risk biomarker. Results are consistent with all experimental and previous clinical and epidemiologic data. Results further implicate NNK as an independent etiologic risk factor in lung cancer.

53 Cigarette Smoke Constituents Targeted for Regulation by WHO, and Their Biomarkers Benzo[a]pyrene NNK, NNN Acrolein Benzene 1,3-Butadiene Carbon monoxide Acetaldehyde Formaldehyde Nicotine 1-HOP or PheT in urine NNAL and NNN in urine 3-HPMA in urine SPMA in urine MHBMA in urine Exhaled CO Leukocyte DNA adducts Nicotine metabolites D.M. Burns et al. Tob. Control. 17: 132-141 (2008); S.S. Hecht, Carcinogenesis 23: 907-922 (2002); L. Chen et al. Chem. Res. Toxicol. 20: 108-133 (2007); S.G. Carmella et al. Chem. Res. Toxicol., 22: 734-741 (2009)

54 Outline of Presentation Tobacco and cancer Background on tobacco carcinogen biomarkers Examples of tobacco carcinogen biomarkers –Total NNAL –Formaldehyde-DNA adducts

55 Formaldehyde – Genetic Toxicology Genotoxic –Mutagenic –DNA protein cross-links –DNA strand breaks –Sister chromatid exchanges –Chromosomal aberrations These changes initiated by reactions with DNA to form adducts No previous reports of formaldehyde DNA adducts in humans International Agency for Research on Cancer Monographs, Volume 88 (2006)

56 Structures of Formaldehyde-DNA Adducts R. Shapiro et al. (1980); F. Beland et al. (1984)

57 Conversion of N 6 -HOMe-dAdo to N 6 -Me-dAdo M. Wang et al. Chem. Res. Toxicol. 20: 1141-1148 (2007)

58 Outline of Analytical Method for N 6 -HOMe-dAdo in DNA

59 LC-ESI-MS/MS-SRM Chromatograms of N 6 -Me-dAdo in Smokers' Leukocyte DNA

60 Typical LC-ESI-MS/MS-SRM Chromatograms of N 6 -Me-dAdo in Leukocyte DNA

61 Levels of N 6 -HOMe-dAdo (as N 6 -Me-dAdo) in Leukocyte DNA of Smokers and Non-smokers

62 Previous Studies of Leukocyte DNA Adducts in Smokers vs. Non-smokers Most used 32 P-postlabelling and immunoassay – inconsistent results comparing smokers and non-smokers Mixed results in 8-OH-dG analyses Marginally higher levels of acetaldehyde-DNA adducts in smokers before stopping Significant difference in BPDE-DNA adducts (2 per 10 8 vs 1 per 10 8 nucleotides) Our results: 5 per 10 8 vs. 0.5 per 10 8 nucleotides IARC Monographs, Vol 83 (2004); Pavanello et al. Mutat. Res 611: 54 (2006); Chen et al Chem. Res. Toxicol. 20: 108 (2007)

63 Sources of Formaldehyde-DNA Adducts Formaldehyde itself in cigarette smoke, but blood levels were not elevated in volunteers exposed to similar amounts Smoking effect on endogenous metabolism Released as a metabolite from nicotine, NNK, or related compounds Transfer from formaldehyde-histone adducts Secondary metabolite from lipid peroxidation or inflammation caused by smoking

64 Conclusions – Formaldehyde DNA Adducts First study to detect formaldehyde-DNA adducts in humans Highly significant differences between smokers and non-smokers Results indicate a previously unrecognized and potentially important role for formaldehyde in smoking-induced cancer M. Wang, G. Cheng, S.S. Hecht et al. Cancer Res., 69: 7170 (2009)

65 Overall Goal Elucidate mechanisms of tobacco-induced cancer and apply this knowledge to cancer prevention.

66 We are Making Progress in Tobacco Control Smoke-free legislation Increased taxation Aggressive anti-tobacco advertising

67 Age-Adjusted Total U.S. Mortality Rates for Lung and Bronchus Cancer Source: SEER data http://seer.cancer.gov/faststats/

68

69 Dorothy Hatsukami

70 Acknowledgements Hecht Lab –Steven Carmella –Mingyao Wang –Irina Stepanov –Pramod Upadhyaya –Brad Hochalter –Silvia Balbo –Shaomei Han –Menglan Chen –Guang Cheng –Lei Meng –Yan Zhong –Aleks Knezevich –John Muzic Core Facilities –Pete Villalta –Chap Le –Xianghua Luo –Yan Zhang –Bruce Lindgren Dorothy Hatsukami –Joni Jensen –Amanda Anderson –Rachel Feuer Tim Church Kristin Anderson Mindy Geisser Jian-Min Yuan Mimi Yu

71 Research Support National Cancer Institute National Institute of Environmental Health Sciences National Institute on Drug Abuse/NCI/NIAAA (TTURC) American Cancer Society Wallin Chair in Cancer Prevention


Download ppt "Tobacco Carcinogen and Toxicant Biomarkers Stephen S. Hecht, Ph.D. Masonic Cancer Center University of Minnesota."

Similar presentations


Ads by Google