Fritz F. Parl, M.D., Ph.D.
Prof Pathology Dir Clinical Chem
Vanderbilt University Medical Center
4605 The Vanderbilt Clinic
Nashville, TN 37232-2562
Breast cancer; molecular genetics; estrogens and estrogen receptor;
Mammary estrogen metabolism leads to the formation of catechol estrogens, the 2-OH and 4-OH derivatives of 17b- estradiol (E2) and estrone (E1). Catechol estrogens have been implicated to play a causative role in breast cancer because they can directly or indirectly induce DNA damage. However, given the shared exposure of all women to estrogens and thereby to the potentially carcinogenic catechol estrogens, it is unclear how to explain differences in breast cancer risk between individuals. In normal breast tissue and breast cancer, catechol estrogens are produced by cytochromes P450 1A1 (CYP1A1) and 1B1 (CYP1B1) and inactivated by catechol-O-methyltransferase (COMT). Since the CYP1A1, CYP1B1, and COMT genes exist as wild type and polymorphic variants, we hypothesize that the inherited enzyme variants hold the potential to define differences in catechol estrogen/carcinogen exposure and thereby explain differences in breast cancer risk. In order to test this hypothesis, we will pursue the following specific aims. Aim 1. To determine whether the amino acid substitutions in variant CYP1B1 affect estrogen metabolism. To this end, we will perform a systematic analysis of each of the four common polymorphisms in exon 2 (codon 48:Arg & Gly; codon 119:Ala & Ser) and exon 3 (codon 432:Val & Leu; codon 453:Asn & Ser) individually and in combination. Each recombinant variant protein will be expressed in E. coli DH5aF''Iq and purified by Ni-chelate affinity chromatography using an N-terminal hexahistidine tag. We will carry out in vitro reconstitution assays combining recombinant purified CYP1B1 and NADPH-P450 reductase with E2 and E1 as substrates. Estrogen metabolites will be extracted and trimethylsilyl derivatives prepared for GC/MS analysis with quantitation by the stable isotope dilution method. These experiments will allow precise analysis of 4-OH and 2-OH catechol estrogen levels and 4-OH/2-OH ratios and define the catalytic effect (Km, Vmax) of each CYP1B1 variant on oxidative estrogen metabolism. Aim 2. To determine whether the amino acid substitutions in variant CYP1A1 and COMT affect estrogen metabolism. Variants of CYP1A1 (codons 171Leu & Ile, 461Thr & Asn, 462Ile & Val) and COMT (codon 158Val & Met) will be expressed, purified, and analyzed similar to CYP1B1 variants. Aim 3. To examine whether the in vitro interaction of CYP1A1, CYP1B1, and COMT variants affects estrogen metabolism. We will compare 4-OH and 2-OH catechol estrogen levels and 4-OH/2-OH ratios produced by wild type CYP1A1, CYP1B1, and COMT with those of variant CYP1A1, CYP1B1, and COMT genotype combinations identified in Aims 1 and 2 to deviate significantly from wild type activity. Aim 4. To determine whether the CYP1A1, CYP1B1, and COMT variants affect in vivo estrogen metabolism in human breast cancer cell lines, normal breast tissue and breast cancer biopsies. Guided by the results of the in vitro assays, we will quantitate 4-OH and 2-OH catechol estrogen levels and 4-OH/2-OH ratios in ten different human breast cancer cell lines, for which we already determined the CYP1B1 and COMT genotypes. Enzyme induction experiments with 2,3,7,8- tetrachlorodibenzo-p-dioxin (TCDD), 12-O- tetradecanoylphorbol-13-acetate (TPA), and indolo[3,2-b] carbazole (ICZ) will answer the question whether the level of expression of variant forms of enzyme differs from that of wild type due to differences in inducibility. The final test of the hypothesis will consist of quantitation of 4-OH and 2-OH catechol estrogen levels and 4-OH/2-OH ratios in 25 - 30 breast tissues (benign and malignant) selected by CYP1A1, CYP1B1, and COMT genotyping. This project will define certain variant CYP1A1, CYP1B1, and COMT genotypes that, alone or in combination, are associated with higher life-long exposure to catechol estrogens and thereby potentially increased risk of breast cancer. .
- Parl FF, Dawling S, Roodi N, Crooke PS "Estrogen metabolism and breast cancer. A risk model." AA NY Acad Sci, 115568-75, 2009.
- Parl FF "A need for true GSTM1 and GSTT1 genotyping." Cancer Epidemiol Biomarkers Prev, 182793, 2009.
- Parl FF, Egan KM, Li C, Crooke PS "Estrogen exposure, metabolism, and enzyme variants in a model for breast cancer risk prediction." Cancer Informatics, 7109-121, 2009.
- Bradley, C, van der Meer, R, Roodi, N, Yan, H, Chandrasekharan, MB, Sun, ZW, Mernaugh, RL, Parl, FF Carcinogen-induced histone alteration in normal human mammary epithelial cells. Carcinogenesis, 28(10), 2184-92, 2007.
- Belous, AR, Hachey, DL, Dawling, S, Roodi, N, Parl, FF Cytochrome P450 1B1-mediated estrogen metabolism results in estrogen-deoxyribonucleoside adduct formation. Cancer Res, 67(2), 812-7, 2007.
- Crooke, PS, Ritchie, MD, Hachey, DL, Dawling, S, Roodi, N, Parl, FF Estrogens, enzyme variants, and breast cancer: a risk model. Cancer Epidemiol Biomarkers Prev, 15(9), 1620-9, 2006.
- Parl, FF Glutathione S-transferase genotypes and cancer risk. Cancer Lett, 221(2), 123-9, 2005.
- McLaren, BK, Gobbi, H, Schuyler, PA, Olson, SJ, Parl, FF, Dupont, WD, Page, DL Immunohistochemical expression of estrogen receptor in enlarged lobular units with columnar alteration in benign breast biopsies: a nested case-control study. Am J Surg Pathol, 29(1), 105-8, 2005.
- Canter, JA, Kallianpur, AR, Parl, FF, Millikan, RC Mitochondrial DNA G10398A polymorphism and invasive breast cancer in African-American women. Cancer Res, 65(17), 8028-33, 2005.
- Roodi, Nady, Dupont, William D, Moore, Jason H, Parl, Fritz F Association of homozygous wild-type glutathione S-transferase M1 genotype with increased breast cancer risk. Cancer Res, 64(4), 1233-6, 2004.
- Dawling, S, Hachey, DL, Roodi, N, Parl, FF In vitro model of mammary estrogen metabolism: structural and kinetic differences between catechol estrogens 2- and 4-hydroxyestradiol. Chem Res Toxicol, 17(9), 1258-64, 2004.
- Kallianpur, Asha R, Hall, Lynn D, Yadav, Meeta, Christman, Brian W, Dittus, Robert S, Haines, Jonathan L, Parl, Fritz F, Summar, Marshall L Increased prevalence of the HFE C282Y hemochromatosis allele in women with breast cancer. Cancer Epidemiol Biomarkers Prev, 13(2), 205-12, 2004.
- Dawling, Sheila, Roodi, Nady, Parl, Fritz F Methoxyestrogens exert feedback inhibition on cytochrome P450 1A1 and 1B1. Cancer Res, 63(12), 3127-32, 2003.
- Parl, F F Multiple mechanisms of estrogen receptor gene repression contribute to ER-negative breast cancer. Pharmacogenomics J, 3(5), 251-3, 2003.
- Hachey, David L, Dawling, Sheila, Roodi, Nady, Parl, Fritz F Sequential action of phase I and II enzymes cytochrome p450 1B1 and glutathione S-transferase P1 in mammary estrogen metabolism. Cancer Res, 63(23), 8492-9, 2003.
- Dawling, S, Roodi, N, Mernaugh, R L, Wang, X, Parl, F F Catechol-O-methyltransferase (COMT)-mediated metabolism of catechol estrogens: comparison of wild-type and variant COMT isoforms. Cancer Res, 61(18), 6716-22, 2001.
- Garte, S, Gaspari, L, Alexandrie, A K, Ambrosone, C, Autrup, H, Autrup, J L, Baranova, H, Bathum, L, Benhamou, S, Boffetta, P, Bouchardy, C, Breskvar, K, Brockmoller, J, Cascorbi, I, Clapper, M L, Coutelle, C, Daly, A, Dell'Omo, M, Dolzan, V, Dresler, C M, Fryer, A, Haugen, A, Hein, D W, Hildesheim, A, Hirvonen, A, Hsieh, L L, Ingelman-Sundberg, M, Kalina, I, Kang, D, Kihara, M, Kiyohara, C, Kremers, P, Lazarus, P, Le Marchand, L, Lechner, M C, van Lieshout, E M, London, S, Manni, J J, Maugard, C M, Morita, S, Nazar-Stewart, V, Noda, K, Oda, Y, Parl, F F, Pastorelli, R, Persson, I, Peters, W H, Rannug, A, Rebbeck, T, Risch, A, Roelandt, L, Romkes, M, Ryberg, D, Salagovic, J, Schoket, B, Seidegard, J, Shields, P G, Sim, E, Sinnet, D, Strange, R C, StÃ¼cker, I, Sugimura, H, To-Figueras, J, Vineis, P, Yu, M C, Taioli, E Metabolic gene polymorphism frequencies in control populations. Cancer Epidemiol Biomarkers Prev, 10(12), 1239-48, 2001.
- Ritchie, M D, Hahn, L W, Roodi, N, Bailey, L R, Dupont, W D, Parl, F F, Moore, J H Multifactor-dimensionality reduction reveals high-order interactions among estrogen-metabolism genes in sporadic breast cancer. Am J Hum Genet, 69(1), 138-47, 2001.
- Carter, B A, Page, D L, Schuyler, P, Parl, F F, Simpson, J F, Jensen, R A, Dupont, W D No elevation in long-term breast carcinoma risk for women with fibroadenomas that contain atypical hyperplasia. Cancer, 92(1), 30-6, 2001.
- Nass, S J, Herman, J G, Gabrielson, E, Iversen, P W, Parl, F F, Davidson, N E, Graff, J R Aberrant methylation of the estrogen receptor and E-cadherin 5' CpG islands increases with malignant progression in human breast cancer. Cancer Res, 60(16), 4346-8, 2000.
- Hanna, I H, Dawling, S, Roodi, N, Guengerich, F P, Parl, F F Cytochrome P450 1B1 (CYP1B1) pharmacogenetics: association of polymorphisms with functional differences in estrogen hydroxylation activity. Cancer Res, 60(13), 3440-4, 2000.
- Parl FF "Estrogens, Estrogen Receptor and Breast Cancer." IOS Press, Amsterdam, Oxford, Tokyo, 2000.
- Dupont, W D, Page, D L, Parl, F F, Plummer, W D, Schuyler, P A, Kasami, M, Jensen, R A Estrogen replacement therapy in women with a history of proliferative breast disease. Cancer, 85(6), 1277-83, 1999.
- Bailey, L R, Roodi, N, Dupont, W D, Parl, F F Association of cytochrome P450 1B1 (CYP1B1) polymorphism with steroid receptor status in breast cancer. Cancer Res, 58(22), 5038-41, 1998.
- Bailey, L R, Roodi, N, Verrier, C S, Yee, C J, Dupont, W D, Parl, F F Breast cancer and CYPIA1, GSTM1, and GSTT1 polymorphisms: evidence of a lack of association in Caucasians and African Americans. Cancer Res, 58(1), 65-70, 1998.
- Lapidus, R. G., Nass, S. J., Butash, K. A., Parl, F. F., Weitzman, S. A., Graff, J. G., Herman, J. G., Davidson, N. E "Mapping of ER gene CpG island methylation by methylation specific PCR." Cancer Res , 582515-2519, 1998.
- Verrier, C S, Roodi, N, Yee, C J, Bailey, L R, Jensen, R A, Bustin, M, Parl, F F High-mobility group (HMG) protein HMG-1 and TATA-binding protein-associated factor TAF(II)30 affect estrogen receptor-mediated transcriptional activation. Mol Endocrinol, 11(8), 1009-19, 1997.
- Benge, H, Bodor, G S, Younger, W A, Parl, F F Impact of managed care on the economics of laboratory operation in an academic medical center. Arch Pathol Lab Med, 121(7), 689-94, 1997.
- Lapidus, R G, Ferguson, A T, Ottaviano, Y L, Parl, F F, Smith, H S, Weitzman, S A, Baylin, S B, Issa, J P, Davidson, N E Methylation of estrogen and progesterone receptor gene 5' CpG islands correlates with lack of estrogen and progesterone receptor gene expression in breast tumors. Clin Cancer Res, 2(5), 805-10, 1996.
- Yaich LE, Roodi N, Bailey LR, Verrier CS, Yee CJ, Cavener DR, Parl FF "Analysis of estrogen receptor gene, transcript and protein in ER-positive and -negative breast cancer cell lines." Endocrine-Related Cancer, 2293-309, 1995.
- Roodi, N, Bailey, L R, Kao, W Y, Verrier, C S, Yee, C J, Dupont, W D, Parl, F F Estrogen receptor gene analysis in estrogen receptor-positive and receptor-negative primary breast cancer. J Natl Cancer Inst, 87(6), 446-51, 1995.
- Dupont, W D, Page, D L, Parl, F F, Vnencak-Jones, C L, Plummer, W D, Rados, M S, Schuyler, P A Long-term risk of breast cancer in women with fibroadenoma. N Engl J Med, 331(1), 10-5, 1994.
- Ottaviano YL, Issa JP, Parl FF, Smith HS, Baylin SB, Davidson NE "Methylation of the estrogen receptor gene CpG island marks loss of estrogen receptor expression in human breast cancer cells." Cancer Res, 542552-2555, 1994.
- Yee, C J, Roodi, N, Verrier, C S, Parl, F F Microsatellite instability and loss of heterozygosity in breast cancer. Cancer Res, 54(7), 1641-4, 1994.
- Caleffi, M, Teague, M W, Jensen, R A, Vnencak-Jones, C L, Dupont, W D, Parl, F F p53 gene mutations and steroid receptor status in breast cancer. Clinicopathologic correlations and prognostic assessment. Cancer, 73(8), 2147-56, 1994.
- Dupont, W D, Parl, F F, Hartmann, W H, Brinton, L A, Winfield, A C, Worrell, J A, Schuyler, P A, Plummer, W D Breast cancer risk associated with proliferative breast disease and atypical hyperplasia. Cancer, 71(4), 1258-65, 1993.
- Yaich, L, Dupont, W D, Cavener, D R, Parl, F F Analysis of the PvuII restriction fragment-length polymorphism and exon structure of the estrogen receptor gene in breast cancer and peripheral blood. Cancer Res, 52(1), 77-83, 1992.
- Foster, B D, Cavener, D R, Parl, F F Binding analysis of the estrogen receptor to its specific DNA target site in human breast cancer. Cancer Res, 51(13), 3405-10, 1991.
- Parl, F F, Cavener, D R, Dupont, W D Genomic DNA analysis of the estrogen receptor gene in breast cancer. Breast Cancer Res Treat, 14(1), 57-64, 1989.
- Dupont, W D, Page, D L, Rogers, L W, Parl, F F Influence of exogenous estrogens, proliferative breast disease, and other variables on breast cancer risk. Cancer, 63(5), 948-57, 1989.
- Parl, F F, Posey, Y F Discrepancies of the biochemical and immunohistochemical estrogen receptor assays in breast cancer. Hum Pathol, 19(8), 960-6, 1988.
- Parl, F F, Schonbaum, C P, Cox, D L, Cavener, D R Detection of estrogen receptor mRNA in human uterus. Mol Cell Endocrinol, 52(3), 235-42, 1987.
- Parl, F F, Wetherall, N T, Halter, S, Schuffman, S, Mitchell, W M Comparison of histochemical and biochemical assays for estrogen receptor in human breast cancer cell lines. Cancer Res, 44(1), 415-21, 1984.
- Parl, F F Estrogen receptor determination in human breast cancer. Prog Clin Pathol, 9155-72, 1984.
- Parl, F F, Schmidt, B P, Dupont, W D, Wagner, R K Prognostic significance of estrogen receptor status in breast cancer in relation to tumor stage, axillary node metastasis, and histopathologic grading. Cancer, 54(10), 2237-42, 1984.
- Parl, F F, Richardson, L D The histologic and biologic spectrum of tubular carcinoma of the breast. Hum Pathol, 14(8), 694-8, 1983.
- Parl, F F, Dupont, W D A retrospective cohort study of Histologic risk factors in breast cancer patients. Cancer, 50(11), 2410-6, 1982.
- Parl, F F, Wagner, R K The histopathological evaluation of human breast cancers in correlation with estrogen receptor values. Cancer, 46(2), 362-7, 1980.
- Entenmann, A H, Parl, F F, Jungblut, P W Studies on the involvement of lysosomes in estrogen action, II. Seasonal variation in the sedimentation patterns of endometrial lysosomes from prepuberal pigs. Hoppe Seylers Z Physiol Chem, 360(11), 1651-5, 1979.
- M.D. - University of Gottingen, 1968
- Internship - Mercer Medical Center
- Post Graduate Training - New York University Medical Center
- Post Graduate Training - Rudolf Virchow Hospital
- Residency - New York University Medical Center