|Partner Organization||Partner Country|
|Université Paris-Saclay (INRA)||France|
|Berlin Institute of Health (BIH)||Germany|
The main objectives of the OCTOPUS project were:
- to determine, in mouse models, to what extent apoA-I/HDL modulate gut microbiota composition, intestinal homeostasis/immunity, plasma metabolome and atherosclerosis development.
- to determine whether apoA-I/HDL levels modulate gut microbiota composition and plasma metabolome in human cohorts, and to identify possible correlations between microbiota composition and metabolic parameters.
- to conduct SNPs analyses to highlight polymorphisms in genes involved in the microbiota-host cross-talk which could influence apoA-I/HDL levels.
- to disseminate to the general public, scientific community, and stakeholders the scientific questions, aims, initiatives and results of the project.
The results obtained in mouse models confirm the protective role of apoA-I/HDL against atherosclerosis. However, HDL seems to be mandatory for the TMAO-mediated deleterious effects on atherosclerosis. Those effects do not seem to involve an increased activation of the immune system, nor macroscopic changes in the morphology of lymphoid organs, liver, kidney, intestine, and lungs.
Metabolomic analyses on plasma and hepatic gene expression indicated that increased dietary choline besides elevating TMAO levels also modulates other metabolic pathways, including the methionine cycle leading to altered homocysteine concentration.
Data obtained in germfree mice that received the cecal microbiota from donor mice with varying levels of HDL and fed diets with different choline content showed that the microbiota per se is not able to modulate atherosclerosis development.
The evaluation of fecal microbiota composition showed significant differences between subjects with low or high HDL, but plasma levels of TMAO did not associate with plasma HDL levels. Evaluation of SNPs in candidate genes with a pivotal role in HDL metabolism and gut mucosa-microbiota interaction did not detect the presence of major variants of these genes in the LURIC population. Epigenome wide association studies were performed and significant associations with ApoA-1 were identified, whose most significant was a CpG at the DHCR24 locus.
The dissemination has been pursued throughout the entire project and will continue beyond the project closure. Both general public and peers were approached with focused preparation of social media posts, scientific articles, and sharing of results at scientific conferences.
Expected impacts. Although no individual bacterial species have been identified/isolated that may impact on atherosclerosis development through a modulation of dietary choline metabolization, this project demonstrates with some consistency that varying levels of HDL, both in humans and mice, do not make the gut microbiota more or less pro-atherogenic. At the same time, the project shows that HDL are essential for the pro-atherogenic effects of TMAO to occur, thus a stratification of patients combining HDL levels with TMAO levels can be hypothesized.
The OCTOPUS project was designed and realized with the aim of shedding light on an aspect of apoA-I/HDL pathophysiology that was never properly investigated before: does apoA-I/HDL affect atherosclerosis development, intestinal inflammation and the immunoinflammatory status by modulating the presence of TMAO-producing (or other) bacteria in the gut?
The most relevant results obtained in OCTOPUS project are:
· ApoA-I/HDL are confirmed to be protective against atherosclerosis development in animal models: mice expressing apoA-I have smaller lesion size than mice not expressing apoA-I.
· A high intake of dietary choline results into an increased plasma concentration of TMAO and worsens atherosclerosis development, but only if apoA-I is expressed. In other words, HDL is a prerequisite for this to happen.
· As a consequence, although HDL is per se protective against the development of atherosclerotic lesions, its presence seems to be necessary for TMAO to exhibit its pro-atherogenic effect: mice expressing apoA-I fed low-choline diet develop smaller lesions than mice fed high-choline diet. That either suggests that TMAO may impair the athero-protective activity of HDL, or that HDL may work as TMAO carrier throughout the bloodstream, contributing to the exploitation of its effects.
· The microbiota composition of mice expressing or not apoA-I showed little, yet sizeable, differences. Differences are also found in the gut microbiota of subjects with low or elevated HDL plasma levels. These differences, however, do not make the microbiota any pro- or anti- atherogenic per se, as fecal transplants into atherosclerosis-prone mice failed to show any difference in plaque formation.
|Authors||Title||Year, Issue, PP||Partners Number||Doi|
|Delgado GE*, Krämer BK, Scharnagl H, Fauler G, Stojakovic T, März W, Kleber ME*, Lammert A||Bile Acids in Patients with Uncontrolled Type 2 Diabetes Mellitus - The Effect of Two Days of Oatmeal Treatment.||2020, 128, 624||2||10.1055/a-1069-7330.|
|Delgado GE*, Krämer BK, Siekmeier R, Yazdani B, März W, Leipe J, Kleber ME*||Influence of smoking and smoking cessation on biomarkers of endothelial function and their association with mortality.||2020, 292, 52||2||10.1016/j.atherosclerosis.2019.11.017.|
|Busnelli M*, Manzini S*, Jablaoui A, Bruneau A, Kriaa A, Philippe C, Arnaboldi F, Colombo A, Ferrari B, Ambrogi F*, Maguin E, Rhimi M, Chiesa G*, Gérard P*||Fat-Shaped Microbiota Affects Lipid Metabolism, Liver Steatosis, and Intestinal Homeostasis in Mice Fed a Low-Protein Diet.||2020, 64, e1900835||5||0.1002/mnfr.201900835|
|Berger M, Kleber ME*, Delgado GE*, März W, Andreas M, Hellstern P, Marx N, Schuett KA.||Trimethylamine N-Oxide and Adenosine Diphosphate-Induced Platelet Reactivity Are Independent Risk Factors for Cardiovascular and All-Cause Mortality.||2020, 126, 660||2||10.1161/CIRCRESAHA.119.316214|
|Yazdani B, Kleber ME*, Yücel G, Delgado GE*, Benck U, Krüger B, März W, Krämer BK.||Association of double product and pulse pressure with cardiovascular and all-cause mortality in the LURIC study.||2020, 22, 2332||2||10.1111/jch.14067|
|Delgado GE*, Krämer BK, März W, Hellstern P, Kleber ME*, Leipe J.||Immune Status and Mortality in Smokers, ex-Smokers and never-Smokers: The Ludwigshafen Risk and Cardiovascular Health Study.||2021, 23, 1191||2||10.1093/ntr/ntab011|
|Wallentin L, Eriksson N, Olszowka M, Grammer TB, Hagström E, Held C, Kleber ME*, Koenig W, März W, Stewart RAH, White HD, Åberg M, Siegbahn A||Plasma proteins associated with cardiovascular death in patients with chronic coronary heart disease: A retrospective study.||2021, 18, e1003513||1||10.1371/journal.pmed.1003513|
|Delgado GE*, Kleber ME*, Moissl AP, Yazdani B, Kusnik A, Ebert MP, März W, Krämer BK, Lammert A, Teufel A.||Surrogate scores of advanced fibrosis in NAFLD/NASH do not predict mortality in patients with medium-to-high cardiovascular risk.||2021. 321, G252||2||10.1152/ajpgi.00058.2021|
|Clauss M, Gérard P*, Mosca A, Leclerc M.||Interplay between exercise intensity and gut microbiota in endurance sports: consequences on health and performance.||2021, 8, 637010||1||10.3389/fnut.2021.637010|
|Yazdani B, Delgado GE*, Scharnagl H, Krämer BK, Drexel H, März W, Scherberich JE, Leiherer A, Kleber ME*.||Combined Use of Serum Uromodulin and eGFR to Estimate Mortality Risk.||2021, 8, 723546||2||10.3389/fmed.2021.723546|
|McGranaghan P, Kirwan JA*, Garcia-Rivera MA*, Pieske B, Edelmann F, Blaschke F, Appunni S, Saxena A, Rubens M, Veledar E, Trippel TD.||Lipid metabolite biomarkers in cardiovascular disease: Discovery and biomechanism translation from human studies.||2021, 11, 621||2||10.3390/metabo11090621|
|Juste C, Gérard P*.||Cholesterol-to-Coprostanol Conversion by the Gut Microbiota: What We Know, Suspect, and Ignore.||2021, 9, 1881||1||10.3390/microorganisms9091881|
|Manzini S*, Busnelli M*, Colombo A*, Franchi E*, Grossano P, Chiesa G*.||reString: an open-source Python software to perform automatic functional enrichment retrieval, results aggregation and data visualization.||2021, 11, 23458||5||10.1038/s41598-021-02528-0|
|Schuchardt JP, Cerrato M, Ceseri M, DeFina LF, Delgado GE*, Gellert S, Hahn A, Howard BV, Kadota A, Kleber ME*, Latini R, Maerz W, Manson JE, Mora S, Park Y, Sala-Vila A, von Schacky C, Sekikawa A, Tintle N, Tucker KL, Vasan RS, Harris WS.||Red blood cell fatty acid patterns from 7 countries: Focus on the Omega-3 index.||2022, 179, 102418||2||10.1016/j.plefa.2022.102418|
|Hilvo M, Lääperi M, Jylhä A, Kleber ME*, Hurme R, Scharnagl H, März W, Sinisalo J, Laaksonen R.||Prior myocardial infarction, coronary artery disease extent, diabetes mellitus, and CERT2 score for risk stratification in stable coronary artery disease.||2022, 29, e159||1||10.1093/eurjpc/zwab122|
|Laaksonen J, Mishra PP, Seppälä I, Raitoharju E, Marttila S, Mononen N, Lyytikäinen LP, Kleber ME*, Delgado GE*, Lepistö M, Almusa H, Ellonen P, Lorkowski S, März W, Hutri-Kähönen N, Raitakari O, Kähönen M, Salonen JT, Lehtimäki T.||Mitochondrial genome-wide analysis of nuclear DNA methylation quantitative trait loci.||2022, 31, 1720||2||10.1093/hmg/ddab339|
|Zhang X*, Gérard P*.||Diet-gut microbiota interactions on cardiovascular disease.||2022, 20, 1528||2||10.1016/j.csbj.2022.03.028|
|García-Rivera MA*, Fernández-Ochoa Á, Brüning U, Fritsche-Guenther R, Kirwan JA*.||Identification and validation of small molecule analytes in mouse plasma by liquid chromatography-tandem mass spectrometry: A case study of misidentification of a short-chain fatty acid with a ketone body.||2022, 242, 123298||2||10.1016/j.talanta.2022.123298|
|Busnelli M*, Manzini S*, Colombo A*, Franchi E*, Bonacina F*, Chiara M, Arnaboldi F, Donetti E, Ambrogi F*, Oleari R, Lettieri A, Horner D, Scanziani E, Norata GD*, Chiesa G*.||Lack of apoA-I in apoEKO mice causes skin xanthomas, worsening of inflammation, and increased coronary atherosclerosis in the absence of hyperlipidemia.||2022, 42, 839||8||10.1161/ATVBAHA.122.317790|
|Target group||Authors||Means of communication||Hyperlink|
|Scientists||Graciela Delgado*, Winfried März, Marcus E. Kleber*, Marco Busnelli*, Philippe Gérard*, Olivia Gräbner*, Giulia Chiesa*, A sound microbiota in a sound body through apolipoprotein A-I and HDL: from mouse models to humans (The OCTOPUS Consortium), Biomarkermeeting, Mannheim, 2019||Poster|
|General public||Marco Busnelli*, Giulia Chiesa*, Stefano Manzini*, HDL, aterosclerosi e… microrganismi intestinali: parte uno studio per studiarne lo stretto rapporto, University of Milan RicercaMix Blog, 2019||Blog post|
|Scientists, General Public||Marco Busnelli*, Giulia Chiesa*, Stefano Manzini*, A sound microbiota in a sound body through apolipoprotein A-I and HDL, Hot Nut 2: Hot Topics in Nutrition, Università degli Studi di Milano, Milan, 2019||Talk||Link|
|General Public||Marco Busnelli*, Interview about Hot Nut 2 talk, by https://microbioma.it/, 2019||Talk / Interview||Link|
|Scientists||Stefano Manzini*, Genetic HDL deficiency: insights from a double knock-out mouse model, Rare Disorders of Lipid Metabolism: from Phenotype to Precision Medicine, Università degli Studi di Milano, Milan, 2019||Talk|
|General public||Consortium, Upkeeping of Project Website. Migration of the site to a long-term domain, URL: https://piermatteobaramban.wixsite.com/octopus||Website||Link|
|General public||Consortium, OCTOPUS Facebook social page. The page hosts multilingual (Italian and English) content related to the Project’s activities, with topics ranging from everyday lab work and routine, to the interesting news in the field. Further, Projects meetings and activities are advertised. Date of activity: 12/02/2019 – current Total posts: 88 Followers: 220 Coverage (mean ± SD): 311 ± 342 Clicks (mean ± SD): 42.0 ± 43.2 Highest coverage: 1,753 Highest clicks: 250||Social media||Link|
|General public||Consortium, OCTOPUS Twitter social account. The account hosts multilingual (Italian and English) content related to the Project’s activities, with topics ranging from everyday lab work and routine, to the interesting news in the field. Further, Projects meetings and activities are advertised. Date of activity: 12/02/2019 – current Total tweets: 132 Followers: 20||Social media||Link|
|Scientists||Marco Busnelli*, Stefano Manzini*, Maria Conti, Giulia Chiesa* Impact Of Dietary Choline On Atherosclerosis Development In Conventionally Raised Apoe-KO Mice Expressing Different Levels Of Apoa-I, 87th EAS Congress, Maastricht, 2019||Talk||Link|
|Scientists||Graciela Delgado*, Marcus Kleber*, Influence of apoA-I levels on gene expression and DNA methylation in mice and humans: the OCTOPUS Consortium, Maikammer meeting, Klinischer Lipidstoffwechsel, Maikammer, 2019||Talk|
|University students||Marco Busnelli*: “The importance of gut microbiota”. Academic courses in Toxicology, Biotechnology and Pharmacy. School of Pharmacy – University of Milan, May2020||Lesson|
|Scientists||Stefano Manzini*, Marco Busnelli*, Amin Jablaoui, Aurélia Bruneau, Aïcha Kriaa, Catherine Philippe, Francesca Arnaboldi, Alice Colombo, Benedetta Ferrari, Emmanuelle Maguin, Moez Rhimi, Giulia Chiesa*, Philippe Gérard*: Fat-Shaped Microbiota Affects Lipid Metabolism, Liver Steatosis And Intestinal Homeostasis In Mice Fed A Low-Protein Diet, Fifth SISA Spring Meeting, Virtual, June/July 2020||Poster|
|Scientists||Marco Busnelli*, Stefano Manzini*, Alice Colombo, Giulia Chiesa*: Impact of dietary choline on atherosclerosis development in conventionally raised ApoE-KO mice expressing different levels of ApoA-I Fifth SISA Spring Meeting, Virtual, June/July 2020||Poster|
|Scientists||Mariel A. García-Rivera*, Graciela Delgado*, Winfried März, Marcus E. Kleber*, Marco Busnelli*, Philippe Gérard*, Jennifer Kirwan*, Giulia Chiesa* A sound microbiota in a sound body through apolipoprotein A-I and HDL: from mouse models to humans: the OCTOPUS Consortium. Metabolomics 2020 Online, October 27-29 2020||Poster|
|Scientists||Mariel A. García-Rivera* OCTOPUS: a sound microbiota in a sound body through apolipoprotein A-I and HDL: from mouse models to humans. Postdoc Day 2020, MDC-Berlin, November 13th 2020||Virtual poster|
|General public||Coordinator, HDL e microbiota: un legame per curare l’aterosclerosi Journal: Platinum (Italian version), edited by PubliScoop Più, Il Sole 24 Ore. No. 070; Pub date: 22/03/2021||Press/Magazine||Link|
|General public||Coordinator, HDL and microbiota: a possible link to treat atherosclerosis Journal: Platinum (English version), edited by PubliScoop Più, Il Sole 24 Ore. No. 070; Pub date: 22/03/2021||Press/Magazine||Link|
|Scientists||Gérard P. Role of the gut microbiota in health and disease. How to prove causality ? International Interdisciplinary Colloquium : new challenges induced by microbiomes, Paris, France, 23 March 2021||Talk|
|Scientists||Busnelli M, Manzini S, Colombo A, Fontana F, Oleari R, Potì F, Castiglioni S, Bellosta S, Chiesa G; Gene Expression Profile Reveals That Apoa-I Levels Modulate Inflammation And Lysosomal Activity In Murine Advanced Atherosclerosis, European Atherosclerosis Society (EAS) Congress May 30-June 2, 2021, Helsinki, Finland||Poster|
|Scientists||Manzini S, Busnelli M, Colombo A, Kiamehr M, & Chiesa G; Hands-Down Extraction Of Individual Lipid Moieties From Lipidomics Datasets: Liputils, A Python Module, European Atherosclerosis Society (EAS) Congress May 30-June 2, 2021, Helsinki, Finland||Poster|
|Scientists||Zhang X, Busnelli M, Manzini S, Etienne C, Monnoye M, Dubard A, Tenorio Na, Chiesa G, Gerard P. Emerging Role of the Gut Microbiota in Choline-induced Atherosclerosis through the Modulation of Apolipoprotein A-I/HDL levels , 12th International Symposium On Gut Microbiology, Clermont-Ferrand, France, 23-25 October 2021||Poster|
|Scientists||Zhang X, Busnelli M, Manzini S, Etienne C, Monnoye M, Dubard A, Tenorio Na, Chiesa G, Gerard P. Effet protecteur de l’apolipoprotéine A-I vis-à-vis de l’athérosclérose : rôle du microbiote intestinal ? Journées Francophones de Nutrition, Lille, France, 9-11 November 2021||Poster|
|Scientists||Busnelli M, Franchi E, Manzini S, Colombo A, García-Rivera MA, Kirwan J, Zhang X, Gérard P, G. Chiesa. Dietary choline content and HDL levels contribute to a pro-atherogenic plasma metabolomic profile in mice, 35° Congresso Nazionale Società Italiana per lo Studio dell’Aterosclerosi, virtual, 28–30 novembre 2021||Talk|
|Scientists + General Public||Gérard P. Contribution du microbiote dans le lien Nutrition – Santé : Exemple de deux projets impliquant des transferts de microbiote. Webinar of the Paris Center for Microbiome Medicine (PaCeMM), 13 December 2021||Talk|
|Scientists||Busnelli M, Franchi E, Zhang X, Manzini S, Colombo A, García-Rivera MA, Kirwan J, Gérard P. Chiesa G. Dietary choline content and HDL levels modulate gut microbiota composition and contribute to a pro-atherogenic plasma metabolomic profile in mice. World of Microbiome, 28-30 aprile 2022, Wien, Austria||poster|
|Scientists||Zhang X, Busnelli M, Manzini S, Etienne C, Monnoye M, Dubard A, Tenorio Na, Chiesa G, Gerard P. Protective effect of Apolipoprotein A-I/HDL levels in Choline-induced Atherosclerosis : a role for the gut microbiota ? World of Microbiome, Vienna, Austria, 28-30 April 2022||Poster|
|Scientists||Chiesa G, Manzini S, Colombo A, Franchi E, Garcia-Rivera MA, Kirwan J, Busnelli M; Dietary Choline Supplementation Increases The Plasma Concentration Of Several Amino Acids During Atherosclerosis Development, European Atherosclerosis Society (EAS) Congress May 22-25, 2022, Milan, Italy||Poster|
|Scientists||Franchi E, Busnelli M, Manzini S, Colombo A, Chiesa G; Histological assessment of lipid deposition in peripheral tissues from genetically modified mice with deficiency or overexpression of apoA-I, European Atherosclerosis Society (EAS) Congress, May 22-25, 2022, Milan, Italy||Poster|
|Scientists||Colombo A, Manzini S, Busnelli M, Franchi E, Chiesa G; HDL levels do not impact on the expression of genes playing a pivotal role in intestinal lipid metabolism in apolipoprotein E- knockout mice, European Atherosclerosis Society (EAS) Congress, May 22-25, 2022, Milan, Italy||Poster|
|Scientists||Manzini S, Busnelli M, Colombo A, Franchi E, Grossano P, Chiesa G; Restring: a software to manage functional enrichment of complex experimental designs, European Atherosclerosis Society (EAS) Congress, May 22-25, 2022, Milan, Italy||Poster|
|Scientists||Busnelli M, Franchi E, Colombo A, Manzini, S, Garcia-Rivera MA, Kirwan J, Chiesa G; Dietary choline increases plasma concentration of gut-derived uremic toxins and metabolites associated with chronic kidney disease progression in apoE- knockout mice with elevated HDL; European Atherosclerosis Society (EAS) Congress, May 22-25, 2022, Milan, Italy||Poster|
|Scientists||Colombo A, Busnelli M, Manzini S, Franchi E, Garcia-Rivera MA, Kirwan J, Chiesa G; Impact Of HDL Levels And Dietary Choline Content On Lipid Metabolism And Atherosclerosis Development In Mice, 7th Spring Meeting Giovani Ricercatori (SIIA, SIMI, SIPREC, SISA), June 5-7, 2022, Rimini, Italy||Poster|
|Scientists||Franchi E, Busnelli M, Colombo A, Manzini S, García-Rivera MA, Kirwan J, Zhang X, Gérard P, Chiesa G; Increased Dietary Choline Alters The Plasma Metabolome, Worsens Atherosclerosis Development, But Does Not Make Gut Microbiota Pro-Atherogenic Per Se, 7th Spring Meeting Giovani Ricercatori (SIIA, SIMI, SIPREC, SISA), June 5-7, 2022, Rimini, Italy||Talk|
|Scientists||Franchi E, Busnelli M, Colombo A, Manzini S, García-Rivera MA, Kirwan J, Zhang X, Gérard P, Chiesa G; Increased Dietary Choline Alters The Plasma Metabolome, Worsens Atherosclerosis Development, But Does Not Make Gut Microbiota Pro-Atherogenic Per Se, V Giornata della Ricerca del Centro Enrica Grossi Paoletti, June 17, 2022, Milan, Italy||Talk|
|Patent licence||Partners involved||Year||International eu or national patent||Comment|