José Manuel Fernández-Real, Hospital de Girona “Dr Josep Trueta”.
Institut d’Investigació Biomèdica de Girona
Universitat de Girona i CIBEROBN
Barcelona, 30 de novembre de 2018
Esteatosi hepàtica i el Microbioma
• Introduccció– Com hem arribat aquí?
• Fetge i microbiota
• Microbiota circulant i fibrosi hepàtica
• The FLORINASH study• Troballes clíniques
• Liver transcriptomics-microbiome relationships
• Plasma and urine metabolomics
• Validació a hepatòcits humans i a ratolins
• Fecal microbiota transplantation
• Conclusions
Esteatosi hepàtica i el Microbioma
• Introduccció– Com hem arribat aquí?
• Fetge i microbiota
• Microbiota circulant i fibrosi hepàtica
• The FLORINASH study• Troballes clíniques
• Liver transcriptomics-microbiome relationships
• Plasma and urine metabolomics
• Validació a hepatòcits humans i a ratolins
• Fecal microbiota transplantation
• Conclusions
Esteatosi hepàtica i el Microbioma
Diabetes Care 2011
• Introduccció– Com hem arribat aquí?
• Fetge i microbiota
• Microbiota circulant i fibrosi hepàtica
• The FLORINASH study• Troballes clíniques
• Liver transcriptomics-microbiome relationships
• Plasma and urine metabolomics
• Validació a hepatòcits humans i a ratolins
• Fecal microbiota transplantation
• Conclusions
Esteatosi hepàtica i el Microbioma
Bouter et al. Gastroenterology 20017
J Henao-Mejia et al. Nature , 1-7 (2012) doi:10.1038/nature10809
Increased severity of NASH in Asc- and
Il18-deficient mice is transmissible to co-housed
wild-type animals.
Gut microbiota produce alcohol in patients with NASH
Loomba et al., Cell Metabolism 2017;25(5):1054-1062.
• Introduccció– Com hem arribat aquí?
• Fetge i microbiota
• Microbiota circulant i fibrosi hepàtica
• The FLORINASH study• Troballes clíniques
• Liver transcriptomics-microbiome relationships
• Plasma and urine metabolomics
• Validació a hepatòcits humans i a ratolins
• Fecal microbiota transplantation
• Conclusions
Esteatosi hepàtica i el Microbioma
Clinical need :Biomarkers of liver fibrosis
obese patients FLORINASH cohort
Liver Fibrosis
Biomarkers based on blood microbiota
Blood bacterial taxa assessed by 16S metagenomic
sequencing in buffy coats of control individuals or
patients with liver fibrosis
Lelouvier et al. Hepatology 2016
Blood microbiota sequences
➔ A diagnostic biomarker of the disease
Lelouvier et al. Hepatology 2016
• Introduccció– Com hem arribat aquí?
• Fetge i microbiota
• Microbiota circulant i fibrosi hepàtica
• The FLORINASH study• Troballes clíniques
• Liver transcriptomics-microbiome relationships
• Plasma and urine metabolomics
• Validació a hepatòcits humans i a ratolins
• Fecal microbiota transplantation
• Conclusions
Esteatosi hepàtica i el Microbioma
Nature Medicine 2018
• Morbidly obese non-diabetic patients (metformin-naïve)
• Recruited in Girona, Spain and Rome, Italy
• No antibiotics
• Stable weight for 3 months prior to sampling
• Hepatic biopsy
• Deep clinical phenotyping (OGTT, clamps)
• Introduccció– Com hem arribat aquí?
• Fetge i microbiota
• Microbiota circulant i fibrosi hepàtica
• The FLORINASH study• Troballes clíniques
• Liver transcriptomics-microbiome relationships
• Plasma and urine metabolomics
• Validació a hepatòcits humans i a ratolins
• Fecal microbiota transplantation
• Conclusions
Esteatosi hepàtica i el Microbioma
Age, country and BMI as confounders in FLORINASH
Nature Medicine 2018
Association of Low Microbial Gene Richness and Steatosis
Metagenomic richness decreased according to
degree of liver steatosis
Nature Medicine 2018
Correlations: Bacterial taxa/steatosis
Gram negative bacteria (LPS+) were associated with
liver steatosis
Nature Medicine 2018
Association between liver steatosis,
Microbial Gene Richness (MGR) and metagenomic
Data in obese women
Nature Medicine 2018
Metagenomic biochemical pathways
The metabolisms of LPS, peptidoglycans and branched amino acids
are correlated with steatosis
Nature Medicine 2018
• Introduccció– Com hem arribat aquí?
• Fetge i microbiota
• Microbiota circulant i fibrosi hepàtica
• The FLORINASH study• Troballes clíniques
• Liver transcriptomics-microbiome relationships
• Plasma and urine metabolomics
• Validació a hepatòcits humans i a ratolins
• Fecal microbiota transplantation
• Conclusions
Esteatosi hepàtica i el Microbioma
Liver Transcriptomics: Affected Pathways
Cluster of Insulin resistance,
inflammation, amino acid degradation
especially connected
Nature Medicine 2018
Transcriptomic networks of the liver:
Microbiota filter➔ 2200 genes
➔ Search for « core centers »➔ ~30
Cluster of Insulin resistance, inflammation, amino acid
degradation especially connected
• Introduccció– Com hem arribat aquí?
• Fetge i microbiota
• Microbiota circulant i fibrosi hepàtica
• The FLORINASH study• Troballes clíniques
• Liver transcriptomics-microbiome relationships
• Plasma and urine metabolomics
• Validació a hepatòcits humans i a ratolins
• Fecal microbiota transplantation
• Conclusions
Esteatosi hepàtica i el Microbioma
1H NMR Urine Metabolomics:
Associations with Microbial Gene Richness and Steatosis
Nature Medicine 2018
1H NMR Plasma Metabolomics:
Associations with Microbial Gene Richness and Steatosis
Nature Medicine 2018
Phenylacetate accumulates in plasma with steatosis score
Branched amino acids and hepatic steatosis
Nature Medicine 2018
Clinical and Phenomic ROC Curves
Nature Medicine 2018
Phenome-Wide Crosstalk and Predictive Modelling
Nature Medicine 2018
1. Low Microbial Gene Richness
2. Increased genetic potential for processing of dietary
lipids and endotoxin biosynthesis (Proteobacteria)
3. Hepatic inflammation
4. Dysregulation of BCAA metabolism
5. Increased level of PAA
Les dones amb obesitat i esteatosis mostren :
• Introduccció– Com hem arribat aquí?
• Fetge i microbiota
• Microbiota circulant i fibrosi hepàtica
• The FLORINASH study• Troballes clíniques
• Liver transcriptomics-microbiome relationships
• Plasma and urine metabolomics
• Validació en hepatòcits humans i en ratolins
• Fecal microbiota transplantation
• Conclusions
Esteatosi hepàtica i el Microbioma
Quines són les possibles consequències de
la modificació del microbioma (i els seus
productes) sobre l’expressió de diferents
gens al fetge?
Microbial Phenylacetate (PAA) indueix esteatosi a
Hepatòcits humans
Nature Medicine 2018
Nature Medicine 2018, in press
Microbial Phenylacetate (PAA) indueix esteatosi a
Hepatòcits humans
Microbial Phenylacetate (PAA) indueix esteatosi a
Hepatòcits humans
Nature Medicine 2018
Microbial Phenylacetate (PAA) indueix esteatosi a
Hepatòcits humans
Nature Medicine 2018
PAA Indueix utilització de BCAA a
Hepatocits humans I a ratolins
CTRL PAA
p = 0.0435
Tri
gly
ceri
des (
mg
/g liv
er)
0
2
4
6
CTRL PAA8
10
12
14
16
Uri
nary
Iso
leu
cin
e (A
U)
p = 0.0003
PA
Nature Medicine 2018
• Introduccció– Com hem arribat aquí?
• Fetge i microbiota
• Microbiota circulant i fibrosi hepàtica
• The FLORINASH study• Troballes clíniques
• Liver transcriptomics-microbiome relationships
• Plasma and urine metabolomics
• Validació a hepatòcits humans i a ratolins
• Fecal microbiota transplantation
• Conclusions
Esteatosi hepàtica i el Microbioma
Faecal Microbiota Transplantations Impact the
Steatosis Phenome
Nature Medicine 2018
Faecal Microbiota Transplantations Impact the
Steatosis Phenome
Nature Medicine 2018
Faecal Microbiota Transplantations Impact the
Steatosis Phenome
Nature Medicine 2018
Faecal Microbiota Transplantations Impact the
Steatosis Phenome
Nature Medicine 2018
• Introduction– How we arrived here
• Antecedents
• Circulating microbiota and liver fibrosis
• The FLORINASH study• Clinical findings
• Liver transcriptomics-microbiome relationships
• Plasma and urine metabolomics
• Validation in human hepatocytes and mice
• Fecal microbiota transplantation
• Conclusions
Microbiome in non-alcoholic fatty liver disease
Conclusions
ASSOCIATIVE• Low microbial gene richness associates with steatosis• Increased genetic potential for processing of dietary lipids and
endotoxin biosynthesis (Proteobacteria)• Hepatic inflammation• Dysregulation of AAA and BCAA metabolism• Increased level of PAA
Conclusions
ASSOCIATIVE• Low microbial gene richness associates with steatosis• Increased genetic potential for processing of dietary lipids and
endotoxin biosynthesis (Proteobacteria)• Hepatic inflammation• Dysregulation of AAA and BCAA metabolism• Increased level of PAA
FUNCTIONAL• FMT triggers steatosis and impacts recipient mouse phenome• PAA induces steatosis in primary human hepatocytes and in mice• High amount of shared information content in phenomic and
metagenomic signatures• High predictivity of combined phenomic and metagenomic
signatures
Acknowledgments
University of Rome Tor Vergata
Marina Cardellini, Francesca Davato, Iris Cardolini, Ottavia Porzio, Paolo Gentilieschi, Massimo Federici
INSERM, ToulouseMatteo Serino, Julie Charpentier, Christophe Heymes, Vincent Azalbert, Vincent Blasco-Baque, Frédéric Lopez, Remy Burcelin
University of GironaJèssica Latorre Luque, José Maria Moreno-Navarrete, Josep Puig, Gemma Xifra, Wifredo Ricart, Jose-Manuel Fernandez-Real
Imperial College London
Lesley Hoyles, James Abbott,
Richard Barton, Julien Chilloux,
Antonis Myridakis, Laura Martinez Gili,
Mark Woodbridge, Chris Tomlinson,
Sarah Butcher, Elaine Holmes,
Jeremy Nicholson,
Marc-Emmanuel Dumas
INSERM, Paris
Fabienne Foufelle, Elodie Anthony,
Fadila Rayah,
Catherine Postic
Hospital de Girona (Radiologia)
Dr. Josep Puig
Dr. Gerard Blasco
Dr. Salvador Pedraza
Hospital de Girona (UDEN-TG)
Dr Gemma Xifra
Dr Jose Maria Moreno-Navarrete
Dr Mercé Fernández-Balsells
Dr Francisco Ortega
Oscar Rovira
Maria Arnorriaga
Dr Wifredo Ricart
Acknowledgments
Hospital Tor Vergata de Roma
Dr. Massimo Federici
INSERM, Toulouse
Dr. Rémy Burcelin
Dr. Manuel Portero-Otin
Universitat de Girona I Universitat de Lleida
Dr. Pepus Daunis
Hospital de Bellvitge
Dr. Fernando Fernández-Aranda
Dra. Susana Jiménez
Dr. Oren Contreras