Abstract
Purpose
The gut microbiome is a potentially important contributor to endogenous estrogen levels after menopause. In healthy postmenopausal women, we examined associations of fecal microbiome composition with levels of urinary estrogens, their metabolites, and relevant metabolic pathway ratios implicated in breast cancer risk.
Methods
Eligible postmenopausal women (n = 164) had a body mass index (BMI) ≤ 35 kg/m2 and no history of hormone use (previous 6 months) or cancer/metabolic disorders. Estrogens were quantified in spot urine samples with liquid chromatography-high resolution mass spectrometry (corrected for creatinine). Bacterial DNA was isolated from fecal samples and the V1–V2 hypervariable regions of 16S rRNA were sequenced on the Illumina MiSeq platform. We examined associations of gut microbiome’s indices of within-sample (alpha) diversity (i.e., Shannon, Chao1, and Inverse Simpson), phylogenetic diversity, and the ratio of the two main phyla (Firmicutes and Bacteroidetes; F/B ratio) with individual estrogens and metabolic ratios, adjusted for age and BMI.
Results
In this sample of 164 healthy postmenopausal women, the mean age was 62.9 years (range 47.0–86.0). We found significant inverse associations of observed species with 4-pathway:total estrogens (p = 0.04) and 4-pathway:2-pathway (p = 0.01). Shannon index was positively associated with 2-catechols: methylated 2-catechols (p = 0.04). Chao1 was inversely associated with E1:total estrogens (p = 0.04), and 4-pathway:2-pathway (p = 0.02) and positively associated with 2-pathway:parent estrogens (p = 0.01). Phylogenetic diversity was inversely associated with 4-pathway:total estrogens (p = 0.02), 4-pathway:parent estrogens (p = 0.03), 4-pathway:2-pathway (p = 0.01), and 4-pathway:16-pathway (p = 0.03) and positively associated with 2-pathway:parent estrogens (p = 0.01). F/B ratio was not associated with any of the estrogen measures.
Conclusion
Microbial diversity was associated with several estrogen metabolism ratios implicated in breast cancer risk. Further studies are warranted to confirm these findings in a larger and more representative sample of postmenopausal women, particularly with enrichment of minority participants.
Similar content being viewed by others
Data availability
The microbial RNA sequence data generated and analyzed during the current study are available in the NCBI repository, https://dataview.ncbi.nlm.nih.gov/object/PRJNA905714?reviewer=idghjm9id1npj51bnc8g0bj1pe. The estrogen metabolite data generated during and analyzed during the current study are available from the corresponding author on reasonable request.
References
Grice EA, Segre JA (2012) The human microbiome: our second genome. Annu Rev Genom Hum Genet 13:151–170
Kinross JM, Darzi AW, Nicholson JK (2011) Gut microbiome-host interactions in health and disease. Genom Med 3:14
Sommer F, Backhed F (2013) The gut microbiota–masters of host development and physiology. Nat Rev Microbiol 11:227–238
Ruo SW, Alkayyali T, Win M et al (2021) Role of gut microbiota dysbiosis in breast cancer and novel approaches in prevention, diagnosis, and treatment. Cureus 13:e17472
Eslami-S Z, Majidzadeh-A K, Halvaei S, Babapirali F, Esmaeili R (2020) Microbiome and breast cancer: new role for an ancient population. Front Oncol. https://doi.org/10.3389/fonc.2020.00120
Tzeng A, Sangwan N, Jia M et al (2021) Human breast microbiome correlates with prognostic features and immunological signatures in breast cancer. Genom Med 13:60
Jaye K, Chang D, Li CG, Bhuyan DJ (2022) Gut metabolites and breast cancer: the continuum of dysbiosis, breast cancer risk, and potential breast cancer therapy. Int J Mol Sci 23:9490
Samavat H, Kurzer MS (2015) Estrogen metabolism and breast cancer. Cancer Lett 356:231–243
Tworoger SS, Zhang X, Eliassen AH et al (2014) Inclusion of endogenous hormone levels in risk prediction models of postmenopausal breast cancer. J Clin Oncol 32:3111–3117
Plottel CS, Blaser MJ (2011) Microbiome and malignancy. Cell Host Microbe 10:324–335
Shapira I, Sultan K, Lee A, Taioli E (2013) Evolving concepts: how diet and the intestinal microbiome act as modulators of breast malignancy. ISRN Oncol 2013:693920
Rose DP (1993) Diet, hormones, and cancer. Annu Rev Public Health 14:1–17
Adlercreutz H, Martin F (1980) Biliary excretion and intestinal metabolism of progesterone and estrogens in man. J Steroid Biochem 13:231–244
Fuhrman BJ, Feigelson HS, Flores R et al (2014) Associations of the fecal microbiome with urinary estrogens and estrogen metabolites in postmenopausal women. J Clin Endocrinol Metab 99:4632–4640
Judd HL, Shamonki IM, Frumar AM, Lagasse LD (1982) Origin of serum estradiol in postmenopausal women. Obstet Gynecol 59:680–686
Boyapati SM, Shu XO, Gao YT et al (2004) Correlation of blood sex steroid hormones with body size, body fat distribution, and other known risk factors for breast cancer in post-menopausal Chinese women. Cancer Causes Contr 15:305–311
Beckmann L, Husing A, Setiawan VW et al (2011) Comprehensive analysis of hormone and genetic variation in 36 genes related to steroid hormone metabolism in pre- and postmenopausal women from the breast and prostate cancer cohort consortium (BPC3). J Clin Endocrinol Metab 96:E360–E367
Key TJ, Appleby PN, Reeves GK et al (2015) Steroid hormone measurements from different types of assays in relation to body mass index and breast cancer risk in postmenopausal women: reanalysis of eighteen prospective studies. Steroids 99:49–55
Flores R, Shi J, Fuhrman B et al (2012) Fecal microbial determinants of fecal and systemic estrogens and estrogen metabolites: a cross-sectional study. J Transl Med 10:253
Willett W, Stampfer MJ, Bain C et al (1983) Cigarette smoking, relative weight, and menopause. Am J Epidemiol 117:651–658
Klann E, Williamson JM, Tagliamonte MS et al (2020) Microbiota composition in bilateral healthy breast tissue and breast tumors. Cancer Causes Contr 31:1027–1038
Miller DN, Bryant JE, Madsen EL, Ghiorse WC (1999) Evaluation and optimization of DNA extraction and purification procedures for soil and sediment samples. Appl Environ Microbiol 65:4715–4724
Zoetendal EG, Akkermans AD, De Vos WM (1998) Temperature gradient gel electrophoresis analysis of 16S rRNA from human fecal samples reveals stable and host-specific communities of active bacteria. Appl Environ Microbiol 64:3854–3859
Hamady M, Walker JJ, Harris JK, Gold NJ, Knight R (2008) Error-correcting barcoded primers for pyrosequencing hundreds of samples in multiplex. Nat Methods 5:235–237
Sun Y, Cai Y, Huse SM et al (2012) A large-scale benchmark study of existing algorithms for taxonomy-independent microbial community analysis. Brief Bioinform 13:107–121
Sun Y, Cai Y, Mai V et al (2010) Advanced computational algorithms for microbial community analysis using massive 16S rRNA sequence data. Nucleic Acids Res 38:e205
Cai Y, Sun Y (2011) ESPRIT-Tree: hierarchical clustering analysis of millions of 16S rRNA pyrosequences in quasilinear computational time. Nucleic Acids Res 39:e95
Caporaso JG, Kuczynski J, Stombaugh J et al (2010) QIIME allows analysis of high-throughput community sequencing data. Nat Methods 7:335–336
Pylro VS, Roesch LF, Ortega JM et al (2014) Brazilian microbiome project: revealing the unexplored microbial diversity–challenges and prospects. Microb Ecol 67:237–241
Wang Q, Garrity GM, Tiedje JM, Cole JR (2007) Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl Environ Microbiol 73:5261–5267
Darville LNF, Cline JK, Rozmeski C et al (2020) LC-HRMS of derivatized urinary estrogens and estrogen metabolites in postmenopausal women. J Chromatogr B 1154:122288
MacLean B, Tomazela DM, Shulman N et al (2010) Skyline: an open source document editor for creating and analyzing targeted proteomics experiments. Bioinformatics (Oxford, England) 26:966–968
Miller RC, Brindle E, Holman DJ et al (2004) Comparison of specific gravity and creatinine for normalizing urinary reproductive hormone concentrations. Clin Chem 50:924–932
Shi L, Remer T, Buyken AE, Hartmann MF, Hoffmann P, Wudy SA (2010) Prepubertal urinary estrogen excretion and its relationship with pubertal timing. Am J Physiol-Endocrinol Metabolism 299:E990–E997
Ahn J, Sinha R, Pei Z et al (2013) Human gut microbiome and risk for colorectal cancer. J Natl Cancer Inst 105:1907–1911
Kim BR, Shin J, Guevarra R et al (2017) Deciphering diversity indices for a better understanding of microbial communities. J Microbiol Biotechnol 27:2089–2093
Reese AT, Dunn RR (2018) Drivers of microbiome biodiversity: a review of general rules, feces, and ignorance. MBio 9:e01294-e1318
Faith DP (1992) Conservation evaluation and phylogenetic diversity. Biol Cons 61:1–10
Yaghjyan L, Darville LNF, Cline J et al (2022) Associations of established breast cancer risk factors with urinary estrogens in postmenopausal women. Cancer Causes Contr 33:279–291
Falk RT, Brinton LA, Dorgan JF et al (2013) Relationship of serum estrogens and estrogen metabolites to postmenopausal breast cancer risk: a nested case-control study. Breast Cancer Res 15:R34
Fuhrman BJ, Schairer C, Gail MH et al (2012) Estrogen metabolism and risk of breast cancer in postmenopausal women. J Natl Cancer Inst 104:326–339
Muti P, Bradlow HL, Micheli A et al (2000) Estrogen metabolism and risk of breast cancer: a prospective study of the 2:16alpha-hydroxyestrone ratio in premenopausal and postmenopausal women. Epidemiology 11:635–640
Ho GH, Luo XW, Ji CY, Foo SC, Ng EH (1998) Urinary 2/16 alpha-hydroxyestrone ratio: correlation with serum insulin-like growth factor binding protein-3 and a potential biomarker of breast cancer risk. Ann Acad Med Singapore 27:294–299
Dallal CM, Tice JA, Buist DS et al (2014) Estrogen metabolism and breast cancer risk among postmenopausal women: a case-cohort study within B~FIT. Carcinogenesis 35:346–355
Moore SC, Matthews CE, Ou Shu X et al (2016) Endogenous estrogens, estrogen metabolites, and breast cancer risk in postmenopausal Chinese women. JNCI: J Natl Cancer Inst. https://doi.org/10.1093/jnci/djw103
Eliassen AH, Missmer SA, Tworoger SS, Hankinson SE (2008) Circulating 2-hydroxy- and 16alpha-hydroxy estrone levels and risk of breast cancer among postmenopausal women. Cancer Epidemiol Biomarkers Prev 17:2029–2035
Rinaldi S, Moret CN, Kaaks R et al (2003) Reproducibility over time of measurements of androgens, estrogens and hydroxy estrogens in urine samples from post-menopausal women. Eur J Epidemiol 18:417–424
Travis RC, Key TJ (2003) Oestrogen exposure and breast cancer risk. Breast Cancer Res: BCR 5:239–247
Coburn SB, Stanczyk FZ, Falk RT et al (2019) Comparability of serum, plasma, and urinary estrogen and estrogen metabolite measurements by sex and menopausal status. Cancer Causes Contr: CCC 30:75–86
Onizuka Y, Nagai K, Ideno Y et al (2019) Association between FSH, E1, and E2 levels in urine and serum in premenopausal and postmenopausal women. Clin Biochem 73:105–108
Toniolo PG, Levitz M, Zeleniuch-Jacquotte A et al (1995) A prospective study of endogenous estrogens and breast cancer in postmenopausal women. J Natl Cancer Inst 87:190–197
Missmer SA, Eliassen AH, Barbieri RL, Hankinson SE (2004) Endogenous estrogen, androgen, and progesterone concentrations and breast cancer risk among postmenopausal women. J Natl Cancer Inst 96:1856–1865
Key TJ, Wang DY, Brown JB et al (1996) A prospective study of urinary oestrogen excretion and breast cancer risk. Br J Cancer 73:1615–1619
Onland-Moret NC, Kaaks R, van Noord PA et al (2003) Urinary endogenous sex hormone levels and the risk of postmenopausal breast cancer. Br J Cancer 88:1394–1399
Thomas HV, Reeves GK, Key TJ (1997) Endogenous estrogen and postmenopausal breast cancer: a quantitative review. Cancer Causes Contr 8:922–928
Acknowledgements
This research was supported by pilot funding from the Florida Academic Cancer Center Alliance (FACCA) and the UF Health Cancer Center Bridge Funding. This work has been supported in part by the Tissue Core and the Proteomics & Metabolomics Core facilities at the H. Lee Moffitt Cancer Center & Research Institute, an NCI designated Comprehensive Cancer Center (P30-CA076292). The authors would like to thank Martin Abrams at Moffitt Cancer Center Clinical Laboratory for performing the creatinine assays.
Funding
This research was supported by pilot funding from the Florida Academic Cancer Center Alliance (FACCA) and the UF Health Cancer Center Bridge Funding. This work has been supported in part by the Tissue Core and the Proteomics & Metabolomics Core facilities at the H. Lee Moffitt Cancer Center & Research Institute, an NCI designated Comprehensive Cancer Center (P30-CA076292).
Author information
Authors and Affiliations
Contributions
LY and KE conceived of and designed the study, obtained funding, directed statistical analyses, interpreted results, substantially revised initial drafts of the paper and provided final review and approval. LD, JC and JK analyzed urinary samples to quantify estrogens. MU, VM, ad MT analyzed stool samples for GM. YM and SR recruited participants and collected data and samples. XW performed statistical analyses. LY wrote the first draft of the manuscript which was revised with contribution from all co-authors. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Ethical approval
The study was approved by the Moffitt Cancer Center and UF Institutional Review Boards (UF: IRB 201500572 and IRB 201600709; Moffitt Cancer Center Advarra IRB# Pro00014574). Women were offered a $25 gift card for their participation. The study was performed in accordance with the ethical standards as laid down in the 1964 Declaration of Helsinki and its later amendments.
Informed consent
All participants provided written informed consent.
Research involving human and/or animals participants
The study was approved by the Moffitt Cancer Center and UF Institutional Review Boards (UF: IRB 201500572 and IRB 201600709; Moffitt Cancer Center Advarra IRB# Pro00014574). The study was performed in accordance with the ethical standards as laid down in the 1964 Declaration of Helsinki and its later amendments.
Consent to participate
All participants provided written informed consent.
Consent for publication
Not applicable.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Yaghjyan, L., Mai, V., Darville, L.N.F. et al. Associations of gut microbiome with endogenous estrogen levels in healthy postmenopausal women. Cancer Causes Control 34, 873–881 (2023). https://doi.org/10.1007/s10552-023-01728-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10552-023-01728-5