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Supplementation with Akkermansia muciniphila in overweight and obese human volunteers: a proof-of-concept exploratory study

Abstract

Metabolic syndrome is characterized by a constellation of comorbidities that predispose individuals to an increased risk of developing cardiovascular pathologies as well as type 2 diabetes mellitus1. The gut microbiota is a new key contributor involved in the onset of obesity-related disorders2. In humans, studies have provided evidence for a negative correlation between Akkermansia muciniphila abundance and overweight, obesity, untreated type 2 diabetes mellitus or hypertension3,4,5,6,7,8. Since the administration of A. muciniphila has never been investigated in humans, we conducted a randomized, double-blind, placebo-controlled pilot study in overweight/obese insulin-resistant volunteers; 40 were enrolled and 32 completed the trial. The primary end points were safety, tolerability and metabolic parameters (that is, insulin resistance, circulating lipids, visceral adiposity and body mass). Secondary outcomes were gut barrier function (that is, plasma lipopolysaccharides) and gut microbiota composition. In this single-center study, we demonstrated that daily oral supplementation of 1010 A. muciniphila bacteria either live or pasteurized for three months was safe and well tolerated. Compared to placebo, pasteurized A. muciniphila improved insulin sensitivity (+28.62 ± 7.02%, P = 0.002), and reduced insulinemia (−34.08 ± 7.12%, P = 0.006) and plasma total cholesterol (−8.68 ± 2.38%, P = 0.02). Pasteurized A. muciniphila supplementation slightly decreased body weight (−2.27 ± 0.92 kg, P = 0.091) compared to the placebo group, and fat mass (−1.37 ± 0.82 kg, P = 0.092) and hip circumference (−2.63 ± 1.14 cm, P = 0.091) compared to baseline. After three months of supplementation, A. muciniphila reduced the levels of the relevant blood markers for liver dysfunction and inflammation while the overall gut microbiome structure was unaffected. In conclusion, this proof-of-concept study (clinical trial no. NCT02637115) shows that the intervention was safe and well tolerated and that supplementation with A. muciniphila improves several metabolic parameters.

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Data availability

The data that support the findings of this study are available upon request. All figures are provided with individual values to have a direct access to the raw data. The 16S sequencing datasets generated during the current study are available from the European Genome-Phenome Archive (https://ega-archive.org/) under accession no. EGAS00001003585.

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Acknowledgements

We thank A. Barrois for the excellent technical assistance. We are very grateful to M. Buysschaert for his continuous support and helpful criticism during the preparation of this project. We also thank the volunteers who participated in this study. P.D.C. is a senior research associate at Fonds de la Recherche Scientifique (FRS-FNRS). A.E. is a research associated at FRS-FNRS. C. Druart was supported by a FIRST Spin-Off grant from the Walloon Region (no. 1410053). Research in the Wageningen laboratory of W.M.d.V. was partially supported by a European Research Council (ERC) Advanced Grant no. 250172 (Microbes Inside), the SIAM Gravity Grant no. 024.002.002 and the Spinoza Award of the Netherlands Organization for Scientific Research. P.D.C. is the recipient of grants from the FNRS (nos. J.0084.15 and 3.4579.11) and Projet de Recherche (no. T.0138.14) and FRFS-WELBIO grants (no. WELBIO-CR-2017C-02). This work was supported by the Funds Baillet Latour (Grant for Medical Research 2015), a prize of the Banque Transatlantique Belgium and a FIRST Spin-Off grant from the Walloon Region (no. 1410053). P.D.C. is a recipient of the PoC ERC grant 2016 (no. Microbes4U_713547) and ERC Starting Grant 2013 (no. 336452-ENIGMO). P.D.C. and J.R. are recipients of a grant from the FNRS and FWO (EOS program no. 30770923).

Author information

P.D.C. conceived the project. J.-P.T., M.P.H., A.L., D.M., A.E., C. Depommier, C. Druart, H.P., M.V.H., W.M.d.V. and P.D.C. designed the clinical study. P.D.C. supervised the clinical part of the study and W.M.d.V. contributed to the microbial culturing of A. muciniphila. P.D.C., A.E., C. Depommier, C. Druart, M.d.B., J.-P.T., A.L., D.M. and M.P.H. performed the clinical part of the study. N.M.D. contributed to interpretation of the results. S.V.-S., G.F. and J.R. performed the fecal microbiome sequencing and analysis. P.D.C., A.E. and C. Depommier performed the experiments and interpreted all the results. P.D.C., A.E. and C. Depommier generated the figures and tables. P.D.C. and C. Depommier wrote the manuscript. All authors discussed the results and approved the manuscript.

Correspondence to Patrice D. Cani.

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Competing interests

A.E., C. Druart, H.P., P.D.C. and W.M.d.V. are inventors of patent applications (nos. PCT/EP2013/073972, PCT/EP2016/071327 and PCT/EP2016/060033 filed with the European Patent Office, Australia, Brazil, Canada, China, the Eurasian Patent Organization, Israel, India, Hong Kong, Japan, South Korea, Mexico, New Zealand and the United States) dealing with the use of A. muciniphila and its components in the context of obesity and related disorders. P.D.C. and W.M.d.V. are cofounders of A-Mansia Biotech S.A.

Additional information

Peer Review Information: Joao Monteiro was the primary editor on this article and managed its editorial process and peer review in collaboration with the rest of the editorial team.

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Extended data

Extended Data Fig. 1 Flow chart of the interventional study.

Diagram of the participant selection procedure, which includes the following information: number of individuals enrolled at each step of the study progress; number of individuals included in the final analysis; details of the events that led to a reduction in group size.

Extended Data Fig. 2 Changes in inflammatory parameters and GLP-1.

a, Soluble CD40 Ligand. b, Growth-related oncogene (CXCL1). c, MCP1. d, GLP-1. Differential values (mean difference and mean difference from placebo) are expressed as the mean + s.e.m., either as raw data or as percentages. The bars represent the mean change from baseline value per group, with their s.e.m. Mann–Whitney U-tests or unpaired t-tests were performed to compare the differential values of both treated groups versus the placebo group (intergroup changes), according to the distribution. The respective P values are shown in the table below each plot. The lines represent the raw values before and after 3 months of supplementation. The distribution of values within each group for each timing is illustrated by a box-and-whisker plot. In the box plots, the line in the middle of the box is plotted at the median, and the inferior and superior limits of the box correspond to the 25th and the 75th percentiles respectively. Matched-pairs Wilcoxon signed-rank tests or paired t-tests were performed to verify changes from baseline (intragroup changes), according to the distribution; when drawn, the capped line above the group concerned shows the corresponding P value. Changes between 0 and 3 months across the 3 groups were analyzed with Kruskal–Wallis or one-way ANOVA tests according to the distribution; group-wise comparisons were performed using Bonferroni’s and Tukey’s corrections for multiple testing, respectively. Placebo group, n = 11; pasteurized bacteria group, n = 12; live bacteria group, n = 9 for all parameters except for growth-related oncogene: placebo group, n = 7; pasteurized bacteria group, n = 10; live bacteria group, n = 8. All tests were two-tailed.

Extended Data Fig. 3 Changes in fecal microbiome.

a, Akkermansia muciniphila abundance in feces evaluated by quantitative PCR. Differential values (mean difference and mean difference from placebo) are expressed as the mean ± s.e.m. as raw data. The bars represent the mean change from baseline value per group, with their s.e.m. Mann–Whitney U-tests were performed to compare the differential values of both treated groups versus the placebo group (intergroup changes) according to the distribution. The respective P values are shown in the table below each plot. The lines represent the raw values before and after 3 months of supplementation. The distribution of values within each group for each timing is illustrated by a box-and-whisker plot. In the box plots, the line in the middle of the box is plotted at the median, and the inferior and superior limits of the box correspond to the 25th and the 75th percentiles, respectively. Matched-pairs Wilcoxon signed-rank tests were performed to verify changes from baseline (intragroup changes) according to the distribution. When the difference is significant, a capped line is marked above the group concerned with the corresponding P value. Kruskal–Wallis analyses were used to compare changes between 0 and 3 months across the 3 groups according to the distribution. Placebo group, n = 11; pasteurized bacteria group, n = 12; live bacteria, n = 9. All tests were two-tailed. *P < 0.05. b, Visualization of participants’ fecal microbiota composition at baseline and end point of the intervention. Fecal microbiota dissimilarity between samples is represented by principal coordinates analysis (genus-level Aitchison distance), with six sample groups corresponding to the three different treatment arms at baseline or at end point represented by confidence ellipses (80% confidence interval). Intervention effects are symbolized by the colored arrows, with direction and length corresponding to the shift in group centroid coordinates from baseline to end point for each treatment arm (rescaled ×4 and re-centered at the baseline global centroid). Placebo group, n = 11; pasteurized bacteria group, n = 12; live bacteria, n = 9.

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Fig. 1: Changes in parameters related to glucose metabolism and WBC.
Fig. 2: Changes in parameters related to lipid metabolism.
Fig. 3: Changes in hepatic and general enzymes.
Fig. 4: Changes in anthropometric parameters.
Extended Data Fig. 1: Flow chart of the interventional study.
Extended Data Fig. 2: Changes in inflammatory parameters and GLP-1.
Extended Data Fig. 3: Changes in fecal microbiome.