New Insights on the Structure of the Human Gut Microbiota. Chaysavanh Manichanh, PhD Vall d Hebron Research Institute Barcelona
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1 New Insights on the Structure of the Human Gut Microbiota Chaysavanh Manichanh, PhD Vall d Hebron Research Institute Barcelona Sessio Societat Catalana Malalties Infecciosas i Microbiologia March 20th, 2014
2 HUMAN GUT MICROBIOTA FEI Image Gallery
3 COMPLEXITY OF MICROBIAL COMMUNITIES Complex community Microbial Culture
4 HUMAN MICROBIOME EVOLUTION OF THE TECHNIQUES From culture to High-throughput sequencing Sanger 454 FLX Titanium Microbial Culture before the 90s Illumina (GAIIx) DNA cloning -sequencing Functional genes 16s rrna gene 2000 AB SOLiD Next generation Sequencing
5 10 YEARS BACK Hugenholtz et al., Nature 2008
6 COST OF DNA SEQUENCING Search in PubMed: human gut microbiome --> 2700 hits Current Opinion in Microbiology 2012, 15:
7 CONSORTIUMS
8 META---OMICS Manichanh et al., Nature Rev Gastro Hepato 2012
9 WHO ARE THEY? Oceans: 12 phylas Soil: 20 phylas Qin et al., Nature 2010; Human Microbiome Project, Nature 2012
10 HUMAN MICROBIOME 160 species / individual 18 common species in all individuals (N=124) 57 species in > 90% 75 species in > 75% 3.3 millions non-redundant genes prevalent genes/individual 38% are shared Qin et al., Nature 2010
11 Diversity WHAT ARE THEY DOING? Functions Human Microbiome Project 2012
12 FUNCTIONS Minimal Gut Metagenome: 6000 functions (2000 were annotated) Housekeeping functions (carbon metabolism and amino acid synthesis) Gut specific functions (adhesion and sugar harvesting) Qin et al., Nature 2010
13 FUNCTIONAL BIOMARKERS FOR HOST PROPERTIES Functions that correlate with : Age (12 OGs: starch degradation enzymes increase) Gender (5 OGs: enriched aspartate biosynthesis modules in males) Nations (10 OGs, varies more between than within nationalities) Arumugam et al., Nature 2011
14 COLONIZATION AND EVOLUTION OF THE MICROBIOTA Dominguez-Bello et al., PNAS 2010 We acquire the gut microbiota during delivery. Depending on the delivery mode via the vagina or by c-section, our first microbiota will resemble more to the skin or the vagina population of our mom
15 COLONIZATION AND EVOLUTION OF THE MICROBIOTA
16 ACROSS AGE Comparison between different regions of the world and different culture Great difference between children and adult microbiota during the 3 first years periode After that, the bacterial community evolves towards an adult-like configuration Yatsunenko et al., Nature 2012
17 ACROSS GEOGRAPHY Difference in the phylogenetic microbial composition between individuals living in different countries Yatsunenko et al., Nature 2012
18 MICROBIOME OF THE ELDERLY 160 elderly subjects (> 65 y) vs 9 adults Claesson et al., PNAS 2010
19 GUT MICROBIOTA ENTEROTYPES Arumugam et al., Nature 2011
20 DISEASES ASSOCIATED MICROBIOTA DYSBIOSIS Obesity IBD: Inflammatory Bowel Disease Crohn s disease Ulcerative colitis Complication of cirrhosis IBS: Irritable Bowel syndrome Type 1 diabetes
21 GUT MICROBIOTA AND IBD DYSBIOSIS the same bacterial phyla predominate in the stomach, small intestine, and colon, although their relative abundance and details of the component species vary as a function of position along the gut Diversity reduction in fecal sample Peterson et al., Cell Host Microbe 2008 Manichanh et al., Nature Rev. Gastro Hepatol. 2012
22 FUNCTIONAL BOWEL DISORDERS Diet challenge Diet challenge uncovers a more unstable microbiota in patients Dysbiosis at the species level Manichanh et al., Gut 2013 Bilophila wadsworthia was found positively correlated with volume of gas evacuated in patients at basal diet, r = 0.64; FDR = 0.12
23 GUT MICROBIOME AND OBESITY MOUSE MODEL Turnbaugh et al., 2006; Lean and obese mice are differentiated by their relative abundance of the 2 dominant bacterial divisions: Bacteroidetes and Firmicutes - Obese microbiome has an increase capacity to harvest energy from diet - Transfer of the obese microbiota to germ-free mice increase body fat
24 STUDIES IN HUMAN Turnbaugh et al., 2006; 2009 Lower proportion of Bacteroidetes and a higher proportion of Actinobacteria in obese compared with lean individuals Bacteroidetes: enriched for several carbohydrate metabolism pathways Firmicutes: enriched for transport systems Obesity-associated genes: carbohydrate, lipid and amino-acid metabolism In obese subjects: a reduction of microbial diversity
25 OBESITY - STUDIES IN HUMAN METAHIT Bimodal distribution of microbial genes Low gene count : < 480,000 genes High gene count : > 480,000 genes Species richness between groups of individuals with high and low gene counts 123 non-obese and 169 obese Danish individuals Le Chatelier et al., Nature genera differed significantly in abundance between the HGC and LGC individuals
26 STUDIES IN HUMAN METAHIT Phenotypes of the HGC and LGC groups Le Chatelier et al., Nature 2013
27 STUDIES IN HUMAN METAHIT 58 species significantly different in abundance between LGC and HGC Overall, this suggests that LGC individuals harbour an inflammation-associated microbiota Le Chatelier et al., Nature 2013
28 STUDIES IN HUMAN Diet intervention Diet intervention: 49 obese and overweight subjects 6 weeks: high-protein diet and fibers 6 weeks: body weight stabilization period with 20% increase in total energy Gut microbial composition of LGC (n=18) and HGC (n=27) subjects. Baseline gene count Cotillard et al., Nature 2013
29 STUDIES IN HUMAN Diet intervention HGC LGC Gene richness may help to predict the efficacy of dietary intervention on inflammatory variables in overweight or obese individuals. Cotillard et al., Nature 2013
30 STUDIES IN HUMAN Diet intervention Differences between LGC and HGC subjects in bioclinical variables Cotillard et al., Nature 2013
31 ACKNOWLEDGMENTS Metagenomic LAB: S. Panda A. Santiago X. Martinez S. Filippi D. Campos Digestive Unit Support M. Gallart, E. Varela Clinicians and collaborations: F. Guarner, VHIR, BCN F. Azpiroz, VHIR, BCN I. Bilbao, VHIR, BCN C. Guarner, San Pau, BCN G. Soriano, San Pau, BCN J. Verdaguer, Lleida University J. Dore, Paris, France Bioinformatics: Rob Knight s lab, Colorado MetaHIT consortium IHMS consortium
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