Menopause Is Associated with an Altered Gut Microbiome and Estrobolome, with Implications for Adverse Cardiometabolic Risk in the Hispanic Community Health Study/Study of Latinos

Abstract

Menopause is a pivotal period during which loss of ovarian hormones increases cardiometabolic risk and may also influence the gut microbiome. However, the menopause-microbiome relationship has not been examined in a large study, and its implications for cardiometabolic disease are unknown. In the Hispanic Community Health Study/Study of Latinos, a population with high burden of cardiometabolic risk factors, shotgun metagenomic sequencing was performed on stool from 2,300 participants (295 premenopausal women, 1,027 postmenopausal women, and 978 men), and serum metabolomics was available on a subset. Postmenopausal women trended toward lower gut microbiome diversity and altered overall composition compared to premenopausal women, while differing less from men, in models adjusted for age and other demographic/behavioral covariates. Differentially abundant taxa for post- versus premenopausal women included Bacteroides sp. strain Ga6A1, Prevotella marshii, and Sutterella wadsworthensis (enriched in postmenopause) and Escherichia coli-Shigella spp., Oscillibacter sp. strain KLE1745, Akkermansia muciniphila, Clostridium lactatifermentans, Parabacteroides johnsonii, and Veillonella seminalis (depleted in postmenopause); these taxa similarly differed between men and women. Postmenopausal women had higher abundance of the microbial sulfate transport system and decreased abundance of microbial β-glucuronidase; these functions correlated with serum progestin metabolites, suggesting involvement of postmenopausal gut microbes in sex hormone retention. In postmenopausal women, menopause-related microbiome alterations were associated with adverse cardiometabolic profiles. In summary, in a large U.S. Hispanic/Latino population, menopause is associated with a gut microbiome more similar to that of men, perhaps related to the common condition of a low estrogen/progesterone state. Future work should examine similarity of results in other racial/ethnic groups. IMPORTANCE The menopausal transition, marked by declining ovarian hormones, is recognized as a pivotal period of cardiometabolic risk. Gut microbiota metabolically interact with sex hormones, but large population studies associating menopause with the gut microbiome are lacking. Our results from a large study of Hispanic/Latino women and men suggest that the postmenopausal gut microbiome in women is slightly more similar to the gut microbiome in men and that menopause depletes specific gut pathogens and decreases the hormone-related metabolic potential of the gut microbiome. At the same time, gut microbes may participate in sex hormone reactivation and retention in postmenopausal women. Menopause-related gut microbiome changes were associated with adverse cardiometabolic risk in postmenopausal women, indicating that the gut microbiome contributes to changes in cardiometabolic health during menopause.

Keywords: gut microbiome; menopause.

FIG 1

Postmenopausal gut microbiome is less diverse and more similar to that of men than the premenopausal gut microbiome is to that of men. (a) Overview of study groups, comparisons of interest, and hypotheses. Thickness of black arrows indicates hypothesized relative magnitude of group difference. (b) Least-squares means of the Shannon diversity index in premenopausal (n = 295) and postmenopausal women (n = 1,027) and younger (n = 258) and older men (n = 720). Estimates are from multivariable linear regression, adjusting for age, Hispanic/Latino background, U.S. nativity, AHEI2010, field center, hormonal contraceptive use, antibiotics use in last 6 months, Bristol stool scale, cigarette use, alcohol use, education, income, physical activity, and BMI. (c) Principal coordinate analysis of the Jensen-Shannon divergence. Sample points are colored by study group, and 75% data ellipses are overlaid on the plot. (d) R-squared values from PERMANOVA of the Jensen-Shannon divergence. Group comparisons were obtained using dummy variables and adjusted for the same covariates as in panel b.

FIG 2

Gut microbiome species associated with menopause also differ between men and women. (a) Volcano plots show differentially abundant species for the group comparisons. W statistics are from ANCOM2 analysis adjusting for age, Hispanic/Latino background, U.S. nativity, AHEI2010, field center, hormonal contraceptive use, antibiotics use in last 6 months, Bristol stool scale, cigarette use, alcohol use, education, income, physical activity, and BMI. Effect size (beta) coefficients are from multivariable linear regression on clr-transformed species abundance, adjusting for aforementioned covariates. Effect size represents difference in clr-transformed species abundance between the specified groups. (b) Correlation of effect sizes (beta) for the different group comparisons. Spearman correlation coefficients are displayed on the plots. (c) Heatmap of effect size (beta) for species with detection level of ≥0.6 in ANCOM2 analysis of post- versus premenopausal women. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 in multivariable linear regression.

FIG 3

Menopause is related to gut microbiome functions and deconjugation (estrobolome) orthologs. (a) Heatmap of effect size (beta) for KEGG functional modules with detection level of ≥0.6 in ANCOM2 analysis of post- versus premenopausal women. Effect size (beta) coefficients are from multivariable linear regression on clr-transformed module abundance, adjusting for age, Hispanic/Latino background, U.S. nativity, AHEI2010, field center, hormonal contraceptive use, antibiotics use in last 6 months, Bristol stool scale, cigarette use, alcohol use, education, income, physical activity, and BMI. Effect size represents difference in clr-transformed module abundance between the specified groups. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 in multivariable linear regression. (b) Least-squares means of clr-transformed ortholog abundance in pre- and postmenopausal women and younger and older men. Estimates are from multivariable linear regression, adjusting for the same covariates as in panel a. (c) Partial Spearman’s correlation matrix (age-adjusted) for species, modules, and orthologs, derived from entire study population. Only menopause-related species and modules, and a priori estrobolome orthologs were included in the matrix. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

FIG 4

Sex hormone metabolites are associated with menopause-related gut microbiome species and functions. (a) Least-squares means of inverse normal-transformed metabolite abundance in premenopausal (n = 154) and postmenopausal women (n = 192), adjusting for age, Hispanic/Latino background, U.S. nativity, AHEI2010, field center, hormonal contraceptive use, antibiotics use in last 6 months, Bristol stool scale, cigarette use, alcohol use, education, income, physical activity, and BMI. A total of 28 metabolites (17 androgenic, 7 pregnenolone, and 4 progestin) were tested; shown here are those significantly different between pre- and postmenopausal women in multivariable regression (P < 0.05). (b and c) Partial Spearman’s correlation matrix (age-adjusted) for species, modules, and estrobolome orthologs versus metabolites in premenopausal women (b) and postmenopausal women (c). Only menopause-related metabolites, menopause-related species and modules, and a priori estrobolome orthologs were included in the matrices. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

FIG 5

Association of menopause-related gut microbiome features with continuous cardiometabolic outcomes in postmenopausal women. Beta coefficients for associations of clr-transformed species and functional module abundance with cardiometabolic outcomes (fasting glucose excluding antidiabetic medication users, triglycerides, and HDL cholesterol, excluding lipid-lowering medication users, systolic and diastolic blood pressure excluding antihypertensive medication users, and waist circumference) in postmenopausal women. Estimates are from multivariable linear regression with continuous metabolic indicators as outcomes, adjusting for age, Hispanic/Latino background, U.S. nativity, AHEI2010, field center, hormonal contraceptive use, antibiotics use in last 6 months, Bristol stool scale, cigarette use, alcohol use, education, income, and physical activity (a), with additional adjustment for BMI (b). *, P ≤ 0.05.

FIG 6

Summary of key findings. (a) We found that the effect of sex on the gut microbiome was profoundly greater than the effect of menopause, but both effects were statistically significant. (b) We observed that the postmenopausal gut microbiome was less diverse than the premenopausal gut microbiome and more similar to men than the premenopausal gut microbiome was to men. Further, the postmenopausal microbiome had decreased abundance of microbial β-glucuronidase, indicating reduced estrobolome potential, i.e., deconjugation activity toward sex steroid hormones. (c) Menopause-depleted gut microbiome features were positively associated with progestin metabolites in postmenopausal women, suggesting that the postmenopausal gut microbiome is involved in sex steroid reactivation and retention. (d) Menopause-related gut microbiome changes were associated with an adverse cardiometabolic risk factor profile in postmenopausal women, including lower HDL, higher waist circumference, and higher blood pressure, suggesting that the gut microbiome contributes to menopause-related cardiometabolic risk.