Fecal microbial determinants of fecal and systemic estrogens and estrogen metabolites: a cross-sectional study

Journal of Translational Medicine

James J Goedert⁵  Jacques Ravel⁴  Pawel Gajer⁴  Mitchell H Gail³  Timothy D Veenstra²  Xia Xu²  Barbara Fuhrman¹  Jianxin Shi³  Roberto Flores⁶ 

[1] Hormonal and Reproductive Epidemiology Branch, Division of Cancer Epidemiology and Genetics, National Cancer Institute, Rockville, MD, 20852, USA;Laboratory of Proteomics and Analytical Technologies, Advanced Technology Program, SAIC-Frederick, Frederick, MD, 21702, USA;Biostatistics Branch, Division of Cancer Epidemiology and Genetics, National Cancer Institute, Rockville, MD, 20852, USA;Institute of Genome Sciences, University of Maryland School of Medicine, Baltimore, MD, 21201, USA;Infections and Immunoepidemiology Branch, Division of Cancer Epidemiology and Genetics, National Cancer Institute, 6120 Executive Boulevard, Room 7068, Rockville, MD 20852, USA;Cancer Prevention Fellowship Program, National Cancer Institute, Rockville, MD, 20852, USA

关键词: Estrogen metabolites;    Fecal estrogens;    Postmenopausal estrogens;    β-glucosidase;    β-glucuronidase;    Enterohepatic circulation;    Feces;    Microbiome;   

Others  :  828287
DOI  :  10.1186/1479-5876-10-253

 received in 2012-10-03, accepted in 2012-12-17,  发布年份 2012

【 摘 要 】

Background

High systemic estrogen levels contribute to breast cancer risk for postmenopausal women, whereas low levels contribute to osteoporosis risk. Except for obesity, determinants of non-ovarian systemic estrogen levels are undefined. We sought to identify members and functions of the intestinal microbial community associated with estrogen levels via enterohepatic recirculation.

Methods

Fifty-one epidemiologists at the National Institutes of Health, including 25 men, 7 postmenopausal women, and 19 premenopausal women, provided urine and aliquots of feces, using methods proven to yield accurate and reproducible results. Estradiol, estrone, 13 estrogen metabolites (EM), and their sum (total estrogens) were quantified in urine and feces by liquid chromatography/tandem mass spectrometry. In feces, β-glucuronidase and β-glucosidase activities were determined by realtime kinetics, and microbiome diversity and taxonomy were estimated by pyrosequencing 16S rRNA amplicons. Pearson correlations were computed for each log_e estrogen level, log_e enzymatic activity level, and microbiome alpha diversity estimate. For the 55 taxa with mean relative abundance of at least 0.1%, ordinal levels were created [zero, low (below median of detected sequences), high] and compared to log_e estrogens, β-glucuronidase and β-glucosidase enzymatic activity levels by linear regression. Significance was based on two-sided tests with α=0.05.

Results

In men and postmenopausal women, levels of total urinary estrogens (as well as most individual EM) were very strongly and directly associated with all measures of fecal microbiome richness and alpha diversity (R≥0.50, P≤0.003). These non-ovarian systemic estrogens also were strongly and significantly associated with fecal Clostridia taxa, including non-Clostridiales and three genera in the Ruminococcaceae family (R=0.57−0.70, P=0.03−0.002). Estrone, but not other EM, in urine correlated significantly with functional activity of fecal β-glucuronidase (R=0.36, P=0.04). In contrast, fecal β-glucuronidase correlated inversely with fecal total estrogens, both conjugated and deconjugated (R≤-0.47, P≤0.01). Premenopausal female estrogen levels, which were collected across menstrual cycles and thus highly variable, were completely unrelated to fecal microbiome and enzyme parameters (P≥0.6).

Conclusions

Intestinal microbial richness and functions, including but not limited to β-glucuronidase, influence levels of non-ovarian estrogens via enterohepatic circulation. Thus, the gut microbial community likely affects the risk for estrogen-related conditions in older adults. Understanding how Clostridia taxa relate to systemic estrogens may identify targets for interventions.

Trial registration

Not applicable.

【 授权许可】

   
2012 Flores et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

【 参考文献 】

• [1]Fuhrman BJ, Schairer C, Gail MH, Boyd-Morin J, Xu X, Sue LY, et al.: Estrogen metabolism and risk of breast cancer in postmenopausal women. J Natl Cancer Inst 2012, 104:326-339.
• [2]Key TJ, Appleby PN, Reeves GK, Roddam AW, Helzlsouer KJ, Alberg AJ, et al.: Circulating sex hormones and breast cancer risk factors in postmenopausal women: reanalysis of 13 studies. Br J Cancer 2011, 105:709-722.
• [3]Cleary MP, Grossmann ME: Minireview: Obesity and breast cancer: the estrogen connection. Endocrinology 2009, 150:2537-2542.
• [4]Goderie-Plomp HW, van der Klift M, de Ronde W, Hofman A, de Jong FH, Pols HA: Endogenous sex hormones, sex hormone-binding globulin, and the risk of incident vertebral fractures in elderly men and women: the Rotterdam Study. J Clin Endocrinol Metab 2004, 89:3261-3269.
• [5]Mellstrom D, Vandenput L, Mallmin H, Holmberg AH, Lorentzon M, Oden A, et al.: Older men with low serum estradiol and high serum SHBG have an increased risk of fractures. J Bone Miner Res 2008, 23:1552-1560.
• [6]Lee JS, Lacroix AZ, Wu L, Cauley JA, Jackson RD, Kooperberg C, et al.: Associations of serum sex hormone-binding globulin and sex hormone concentrations with hip fracture risk in postmenopausal women. J Clin Endocrinol Metab 2008, 93:1796-1803.
• [7]Raftogianis R, Creveling C, Weinshilboum R, Weisz J: Estrogen metabolism by conjugation. J Natl Cancer Inst Monogr 2000, 113-124.
• [8]Rose DP: Diet, hormones, and cancer. Annu Rev Public Health 1993, 14:1-17.
• [9]Heimer GM, Englund DE: Enterohepatic recirculation of oestriol studied in cholecystectomized and non-cholecystectomized menopausal women. Ups J Med Sci 1984, 89:107-115.
• [10]Goldin BR, Adlercreutz H, Gorbach SL, Warram JH, Dwyer JT, Swenson L, et al.: Estrogen excretion patterns and plasma levels in vegetarian and omnivorous women. N Engl J Med 1982, 307:1542-1547.
• [11]Plottel CS, Blaser MJ: Microbiome and malignancy. Cell Host Microbe 2011, 10:324-335.
• [12]Rinaldi S, Key TJ, Peeters PH, Lahmann PH, Lukanova A, Dossus L, et al.: Anthropometric measures, endogenous sex steroids and breast cancer risk in postmenopausal women: a study within the EPIC cohort. Int J Cancer 2006, 118:2832-2839.
• [13]Eriksson AL, Lorentzon M, Vandenput L, Labrie F, Lindersson M, Syvanen AC, et al.: Genetic variations in sex steroid-related genes as predictors of serum estrogen levels in men. J Clin Endocrinol Metab 2009, 94:1033-1041.
• [14]Kendall A, Folkerd EJ, Dowsett M: Influences on circulating oestrogens in postmenopausal women: relationship with breast cancer. J Steroid Biochem Mol Biol 2007, 103:99-109.
• [15]Turnbaugh PJ, Hamady M, Yatsunenko T, Cantarel BL, Duncan A, Ley RE, et al.: A core gut microbiome in obese and lean twins. Nature 2009, 457:480-484.
• [16]Flores R, Shi J, Gail MH, Gajer P, Ravel J, Goedert JJ: Assessment of the human faecal microbiota: II. Reproducibility and associations of 16S rRNA pyrosequences. Eur J Clin Invest 2012, 42:855-863.
• [17]Flores R, Shi J, Gail MH, Ravel J, Goedert JJ: ssessment of the human faecal microbiota: I. Measurement and reproducibility of selected enzymatic activities. Eur J Clin Invest 2012, 42:848-854.
• [18]Flores R, Shi J, Gail MH, Gajer P, Ravel J, Goedert JJ: Association of fecal microbial diversity and taxonomy with selected enzymatic functions. PLoS One 2012, 7:e39745.
• [19]Falk RT, Xu X, Keefer L, Veenstra TD, Ziegler RG: A liquid chromatography-mass spectrometry method for the simultaneous measurement of 15 urinary estrogens and estrogen metabolites: assay reproducibility and interindividual variability. Cancer Epidemiol Biomarkers Prev 2008, 17:3411-3418.
• [20]Gallo V, Egger M, McCormack V, Farmer PB, Ioannidis JP, Kirsch-Volders M, et al.: STrengthening the Reporting of OBservational studies in Epidemiology - Molecular Epidemiology (STROBE-ME): an extension of the STROBE statement. Eur J Clin Invest 2012, 42:1-16.
• [21]Goldin BR, Gorbach SL: Effect of Lactobacillus acidophilus dietary supplements on 1,2-dimethylhydrazine dihydrochloride-induced intestinal cancer in rats. J Natl Cancer Inst 1980, 64:263-265.
• [22]Xu X, Veenstra TD, Fox SD, Roman JM, Issaq HJ, Falk R, et al.: Measuring fifteen endogenous estrogens simultaneously in human urine by high-performance liquid chromatography-mass spectrometry. Anal Chem 2005, 77:6646-6654.
• [23]Xu X, Roman JM, Issaq HJ, Keefer LK, Veenstra TD, Ziegler RG: Quantitative measurement of endogenous estrogens and estrogen metabolites in human serum by liquid chromatography-tandem mass spectrometry. Anal Chem 2007, 79:7813-7821.
• [24]Peterson J, Garges S, Giovanni M, McInnes P, Wang L, Schloss JA, et al.: The NIH human microbiome project. Genome Res 2009, 19:2317-2323.
• [25]Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, et al.: QIIME allows analysis of high-throughput community sequencing data. Nat Methods 2010, 7:335-336.
• [26]Edgar RC, Haas BJ, Clemente JC, Quince C, Knight R: UCHIME improves sensitivity and speed of chimera detection. Bioinformatics 2011, 27:2194-2200.
• [27]Cole JR, Wang Q, Cardenas E, Fish J, Chai B, Farris RJ, et al.: The Ribosomal Database Project: improved alignments and new tools for rRNA analysis. Nucleic Acids Res 2009, 37:D141-D145.
• [28]Adlercreutz H, Pulkkinen MO, Hamalainen EK, Korpela JT: Studies on the role of intestinal bacteria in metabolism of synthetic and natural steroid hormones. J Steroid Biochem 1984, 20:217-229.
• [29]Bjornerem A, Emaus N, Berntsen GK, Joakimsen RM, Fonnebo V, Wilsgaard T, et al.: Circulating sex steroids, sex hormone-binding globulin, and longitudinal changes in forearm bone mineral density in postmenopausal women and men: the Tromso study. Calcif Tissue Int 2007, 81:65-72.
• [30]Goodman-Gruen D, Barrett-Connor E: Sex differences in the association of endogenous sex hormone levels and glucose tolerance status in older men and women. Diabetes Care 2000, 23:912-918.
• [31]Chearskul S, Charoenlarp K, Thongtang V, Nitiyanant W: Study of plasma hormones and lipids in healthy elderly Thais compared to patients with chronic diseases: diabetes mellitus, essential hypertension and coronary heart disease. J Med Assoc Thai 2000, 83:266-277.
• [32]Hamady M, Lozupone C, Knight R: Fast UniFrac: facilitating high-throughput phylogenetic analyses of microbial communities including analysis of pyrosequencing and PhyloChip data. ISME J 2010, 4:17-27.
• [33]Jarvenpaa P, Kosunen T, Fotsis T, Adlercreutz H: In vitro metabolism of estrogens by isolated intestinal micro-organisms and by human faecal microflora. J Steroid Biochem 1980, 13:345-349.
• [34]Eriksson H, Gustafsson JA: Excretion of steroid hormones in adults. Steroids in faeces from adults. Eur J Biochem 1971, 18:146-150.
• [35]Adlercreutz H, Martin F, Pulkkinen M, Dencker H, Rimer U, Sjoberg NO, et al.: Intestinal metabolism of estrogens. J Clin Endocrinol Metab 1976, 43:497-505.
• [36]Heimer GM, Englund DE: Enterohepatic recirculation of oestriol: inhibition by activated charcoal. Acta Endocrinol (Copenh) 1986, 113:93-95.
• [37]Wallace BD, Wang H, Lane KT, Scott JE, Orans J, Koo JS, et al.: Alleviating cancer drug toxicity by inhibiting a bacterial enzyme. Science 2010, 330:831-835.
• [38]Dodgson K, White G, Fitzgerald J: Sulfatases of Microbial Origin. 1982.