RELACIÓN ENTRE LA MICROBIOTA INTESTINAL Y LA SALUD Y CONTRIBUCIÓN DE LA FIBRA EN LA DIETA

Transcription

1 RELACIÓN ENTRE LA MICRBITA INTESTINAL Y LA SALUD Y CNTRIBUCIÓN DE LA FIBRA EN LA DIETA Lortorio de Ecologí Microin, Nutrición y Slud. MARINA RMANÍ-PÉREZ

2 1 GUT MICRBITA 2 GUT MICRBITA & DISEASES. BESITY 3 MICRBIME-BASED INTERVENTINS FR PRMTING METABLIC HEALTH. PREBITICS & PRBITICS 4 MICRBITA-BY-DIET INTERACTIN. SCFAs VERVIEW 5 6 GUT MICRBITA-HST CMMUNICATIN GRUP F MICRBIAL ECLGY, NUTRITIN AND HEALTH. n the wy to evlute new proiotics 6.1 Evlution of the potentil proiotic Bcteroides uniformis CECT Evlution of dietry fier supplementtion (WBE) comine with CECT 7771 intervention Lortorio de Ecologí Microin, Nutrición y Slud. MARINA RMANÍ-PÉREZ

3 1. GUT MICRBITA 39 trillion microil cells Body hitt groups 1-3% of ody mss The Humn Microiome Project Consortium HST GENME (Ekryotic) 1 times more cells thn humn cells 15 times more genes thn humn genes HLGENME PHENTYPE PLASTICITY GENME F SYMBITIC MICRBES (Prokryotic)

4 1. GUT MICRBITA SYMBISIS PHYLGENY F BACTERIAL PHYLA HST Actinocteri Firmicutes Clostridium, Enterococcus, Lctocillus nd Ruminococcus Dynmic microil community struture mong individuls Proteocteri Host genome DIET Gender Drugs Antiiotics Stress Aging Delivery mode Bcteroidetes Bcteroides nd Prevotell HST PHENTYPE PLASTICITY

5 1. GUT MICRBITA Stility of metolic pthwys mong individuls Phyl Firmicutes Bcteroidetes Gut microiot HST Energy hrvest Protection ginst pthogens Immunomodultion Energy metolic sttus The Humn Microiome Project Consortium Gut microiot Nutrients Shelter CH metolism Cofctor & Vitmin iosynthesis Gut microiot

6 1. GUT MICRBITA & HST SYMBINTS PATHSYMBINTS SYMBISIS PATHSYMBINTS SYMBINTS HST MUTUALISM Euiosis Dysiosis X CMMENSALISM PARASITISM Immune homeostsis Proinflmmtory stte Helthy stte Intestinl nd extr-inetstinl Inflmmtory DISEASES DC M1 T cells Treg M2 B cells

7 2. GUT MICRBITA & DISEASES Dysiosis PATHSYMBINTS SYMBINTS Proinflmmtory stte CHRNIC Intestinl nd extr-intestinl Inflmmtory DISEASES Inflmmtory owel disese Celic disese Crdiovsculr diseses Mentl diseses (emotionl, cognitive-relted diseses; nxiety, depression, utism, etc) Metolic diseses: Dietes (T1D or T2D), esity, metolic syndorme

8 2. GUT MICRBITA & BESITY prevlence from 1975 (x3) In 216: KEY FACTS F BESITY ADULTS: 39% overweight nd 13% oese. CHILDREN & ADLESCENTS: 18% overweight or oese esity rnks fifth in risk of glol deths in the generl popultion prevlence in low nd middle-income countries BESITY IS PREVENTABLE worldoesity.org

9 Energy intke 2. GUT MICRBITA & BESITY PATHPHYSILGY F BESITY Energy intke Energy expenditure Energy expenditure Stte of Chronic low-grde of inflmmtion X Metolic comoridities T2D, NAFLD, dyslipidemi, crdiovsculr disese, etc Energy imlnce BMI: Kg/m 2 (ody mss index) verweight: 25 BMI 3 esity: BMI 3 diposity METABLIC PRFILE Insulin resistnce Systemic Insulin resistnce

10 2. GUT MICRBITA & BESITY Genetic fctors 3-4% 7-6% Environmentl fctors Gut-rin-periphery xis Periphery Brin Hypothlmus ENERGY IMBALANCE ver-eting diposity Glucose intolernce Insulin resistnce Sedentry life style stiety signls sensitivity DIET: Energy-dense foods Gut microiot Gut Anti-oesity interventions ENERGY BALANCE Restortion of gut-rin communiction EUBISIS

11 2. GUT MICRBITA & BESITY DIET non-digerile polyschrides Gut microil communities Donor Microiot trnsfer Metolic phenotype Energy storge Extrction nd storge of energy from diet +/+ GF Individul predisposition to oesity o/o o/o Gut microiot is n enviromentl fctor of oesity

12 2. GUT MICRBITA & BESITY Len Discordnt Twins GF mice ese Ln GF mice Microiot invsion depends on microiot-y-diet interction LoSF-HiFV HiSF-LoFV x Gut microiot invsion linked to leness Cohousing Ln Ln Bcteroidetes invsion

13 2. GUT MICRBITA & BESITY Ley et l, Nture 26 Actinocteri Proteocteri esity Firmicutes Clostridium, Enterococcus, Lctocillus nd Ruminococcus Mnipultion of microil community structure FAT-R, ft restricted CARB-R, crohydrte restricted Bcteroidetes Bcteroides nd Prevotell MICRBIME-BASED interventions for promoting metolic helth PREBITICS o/o 5% fewer Bcteroidetes PRBITICS FECAL TRANSFER

14 3. MICRBIME-BASED INTERVENTINS FR PRMTING METABLIC HEALTH Microiome-sed interventions Next genertion of Proiotics (Live microorgnisms) Preiotics (Food grde component) Microiot replcement (FMT, microiot community trnsfer) Alter composition/ctivity of host gut microiot Akkermnsi muciniphil cylglycerols Gut rrier integrity Eucterium hllii SCFAs Acette Propionte Butyrte FFAR2, FFAR3 Len helthy donor Helth enefit Metolic Helth Bcteroides cidifciens Mucus lyer GLP-1, PYY? DPPIV GLP-2 Inflmmtion ( INFγ) Secondry BAs TGR5 STα SCFAs? GLP-1, PYY EEC: L cells TGR5 Ft mss AT inflmmtion PPARγ Browning Heptic TG Improved glucose homeostsis Brin Food intke Thermogenesis BAT TGR5 Romní-Pérez M et l, 217

15 3.1 PREBITICS DIETARY FIBER: indigestile plnt polyscchrides NUTRIENT Nturl SURCE NUTRIENT Nturl SURCE Wter-insolule dietry fiers Wter-solule dietry fiers Cellulose Inulin Hemicellulose Cerels Fruit Vegetles Fructooligoscchrides (FS) Chicory root Leek nion Bnn Cerels Brn Legumes Arinoxyln (Hemicellulose) rn ~7% endosperm ~88% Lignin Resistnt strch Cerels Grden ens Arinoxyln oligoscchrides (AXS) Pectin Cerels Whet Non-strch polyscchride (NSP) Corn Whet Brley Fruits Corn flkes Crrots

16 3.2 PRBITICS Strin identifiction Functionl chrcteriztion in vitro & in vivo Sfety ssesment Efficcy Phse 2 humn tril (DBPC). Proiotic vs plceo Effectiveness Phse 3 humn tril Proiotic vs stndrd tretment Clssicl proiotics Currently commercilized proiotics Lctic cid cteri nd ifidocteri Biologicl smples or derived from fermented foods Next Genertion of Proiotics (NGP) Mssive use of DNA sequencing in controlled (16S rrna gene-sed pproch, whole-genome shotgun-sed pproch) epidemiologicl studies NGP: Selected functionlly distinct indigenous cteri with potentil higher efficcy of current proiotic formultion Evlution of the dministrtion of A. muciniphil on prmeters of Metolic Syndorme Proiotic for FD

17 4. MICRBITA-BY-DIET INTERACTIN. SCFAs MICRBITA-BY-DIET INTERACTIN Different diets in humn popultion DIET Microil community structure Burkin Fso children DIET: high in FIBER nd low in ft Europen children DIET: high in niml PRTEIN, SUGAR nd FAT. Low in fier HST Whole grins Dietry fier Xylne, xylose, croxymethylcellulose Metolic phenotype plsticity SCFA-producers Pthogen intestinl microes Benefits to metolic helth De Filippo et l, PNAS 21

18 4. MICRBITA-BY-DIET INTERACTIN. SCFAs DIETARY FIBER Gut microiot metolic routes FERMENTATINS SHRT CHAIN FATTY ACIDS Bcteroides Resistnt strchs (1, 4), rynoxylns, pectin, inulin, FS, mucin, mucopolyscchrides Ríos-Covián et l, 216 & Bsson et l, 216

19 5. GUT MICRBITA-HST CMMUNICATIN SCFAs: ictive molecules GPR41 Gαi Gαi Gαq GPR43 Cellulr response Enteroendocrine cells Neurons Inmmune cells Gut hormones. GLP-1, PYY CLN EECs: L cells Fier Energy homeostsis Inmune homeostsis Colonic trnsit ACh

20 5. GUT MICRBITA-HST CMMUNICATIN IMMUNMDULATIN DC T cells esity Stte of Chronic low-grde of inflmmtion Gut microiot Intestinl immunity M1 M2 Treg B cells Adipose tissue Dysiosis Cytotoxic T cells Th1 nd Th17 B cells incresed M1/M2 rtio IL-6, TNF-α, IL-1β Leptin High ft diet Gut permeility PGN TLRs LPS M cells DNA ND SCFAs High ft diet Insulin resistnce Inflmmtory meditors IFN-γ Systemic Insulin resistnce Metolic endotoxemi Systemic inflmmtion MCHI MCHII CD8+ CD4+ MLN Innte immunity A Adpttive immunity Wire et l. Cell Metolism 216

21 6. GRUP F MICRBIAL ECLGY, NUTRITIN AND HEALTH. n the wy to evlute new proiotics Evlution of proiotics for food Strin identifiction Functionl chrcteriztion in vitro & in vivo Sfety ssesment Phse 2 humn tril (DBPC) Proiotic vs plceo Phse 3 humn tril Proiotic vs stndrd tretment Pthophysiology of metolic nd inflmmtory diseses Proiotic for FD

22 6.1 n the wy to evlute new proiotics in vitro functionl chrcteriztion of Bcteroides uniformis CECT 7771 Bcteroides-Prevotell Bcteroides frgilis Anti-oesity effects Strin identifiction In vitro Functionl chrcteriztion Higher undnce in rest-fed thn in formul-fed infnts. CECT 7771 isolted from stools of helthy rest-fed infnts Mcrophges Different strins of Bcteroides Guffin et l. PLoS ne. 212

23 6.1 n the wy to evlute new proiotics in vivo functionl chrcteriztion of Bcteroides uniformis CECT 7771 In vivo Functionl chrcteriztion of Bcteroides uniformis CE T 7771 IMMUNMETABLIC PARAMETERS C57BL/6 DIET DIET INDUCED BESITY (DI) PRTEINS (% of Kcl) CH (% of Kcl) FAT (% of Kcl) (corn oil) (plm oil) FRUCTSE (% of Kcl) 2 Peyer s Ptches Emnuel Fersni Fecl smples Plceo (1% skimmed milk) Gut microiot CECT 7771 (1X1 9 CFU) Blood Plceo (1% skimmed milk) CECT 7771 (1X1 9 CFU) weeks Triglycerides Cholesterol Glucose Leptin Adipose tissue 1 14 Lympoid cells: B cells nd T cells -Stndrd Diet GTT IMMUNMETABLIC PARAMETERS Myeloid cells: mcrophges Cytokines -High Ft High Fructose diet

24 Cholesterol (mg/dl) Triglycerides (mg/dl) Glycemi (mg/dl) % of Body weight gin Leptin (ng/ml) Glycemi (mg/dl) AUC epwat(g)/1g Bw 6.1 n the wy to evlute new proiotics in vivo functionl chrcteriztion of Bcteroides uniformis CECT 7771 METABLIC PHENTYPE rl glucose intolernce * ** ** ** * plceo DIET Hyperleptinemi, 13 hyperglycemi, 6 hypercholesterolemi, hypertriglyceridemi (corn oil) Len phenotype Adipose tissue plceo PRTEINS (% of Kcl) CH (% of Kcl) 1 DIET 5 c plceo c c plceo c PRTEINS CH FAT (% of Kcl) FRUCTSE (% of Kcl) (% plceo (% of Kcl) (% of Kcl) (plm oil) 2 c (c (p time (min) + HFH + ese phenotype 2 1 plceo plceo

25 Treg cells (%) M1/M2 Treg cells (%) Treg cells (%) B cells (%) cytotoxic T /Helper T B cells (%) M1/M2 cytotoxic T /Helper T B cells (%) cytotoxic T /Helper T 6.1 n the wy to evlute new proiotics in vivo functionl chrcteriztion of Bcteroides uniformis CECT 7771 Mechnistic understnding of how promote nti-oesity effects Adipose tissue Len phenotype DIET PRTEINS (% of Kcl) ANTIINFLAMMATRY RESPNSE CH PRTEINS FAT CH FRUCTSE FAT DIET (% of Kcl) (% of (% Kcl) of Kcl) (% of (% Kcl) of Kcl) (% of Kcl) (corn oil) Blood (corn oil) (plm.5 oil) (plm oil).4.3 FRUCTSE (% of Kcl) 2 DC Cyt T Treg M1/M2 lnce B cells plceo c ese phenotype PRINFLAMMATRY STATE plceo plceo Peyer s Ptches c plceo,c MLN plceo plceo c plceo c Dendritic cells (DC) Cytotoxic T cell (cyt T) regultory T cells (Treg) plceo plceo plceo plceo,c c

26 pg/ml pg/g of tissue pg/g of tissue B cells (%) pg/g of tissue pg/ml B cells (%) pg/ml pg/g of tissue pg/ml B cells (%) pg/ml pg/g of tissue pg/g of tissue 6.1 n the wy to evlute new proiotics in vivo functionl chrcteriztion of Bcteroides uniformis CECT lood TNF ** ** ### plceo plceo plceo IL-1 plceo * ### plceo IL-6 * IL-1 ** ### ### ** plceo IFN ** ** ## IFN ** ## plceo DC Cyt T MLN Treg Peyer s Ptches IL-1 * ### * M1/M2 lnce plceo B cells IFN * EpWAT ### * plceo IL plceo IL-33 plceo ### ### plceo plceo plceo IL-1 ** * ### ANTIINFLAMMATRY STATE Len phenotype DIET ese phenotype PRTEIN (% of Kc PRINFLAMMATRY STATE

27 6.1 n the wy to evlute new proiotics in vivo functionl chrcteriztion of Bcteroides uniformis CECT 7771 TLRs ileum Peyer s Ptches Len phenotype PRTEINS DIET (% of Kcl) (% o TLR plceo plceo Adipose tissue plceo plceo TLR2 TLR4 TLR5 ese phenotype

28 Normlized red counts 6.1 n the wy to evlute new proiotics in vivo functionl chrcteriztion of Bcteroides uniformis CECT 7771 Gut microiot BETA DIVERSITY Gener ALPHA DIVERSITY +B +B

29 6.1 n the wy to evlute new proiotics in vivo functionl chrcteriztion of Bcteroides uniformis CECT 7771 Summry Pro-inflmmtory stte Insulin resistnce oese-like phenotype Anti-inflmmtory effects Insulin sensitivity len-like phenotype M1/M2 TLR5? DC M1 M2 T cells Treg B cells IFN-γ IL-33 IL-1 Peyer s Ptches M1/M2 IFN-γ TLR5 TLR4 TLR2 LPS Butyrte Bcteroides? DIET CECT 7771 PRTEINS (% of Kcl) CH (% of K Helicocter Adipose tissue TNF-α IL-6 IL-33 IL-1 IL-1α IL-1 T5K mice metolic sindrome Vijy-Kumr et l. 21

30 6.1 n the wy to evlute new proiotics in vivo sfety ssesment of Bcteroides uniformis CECT 7771 Sfety ssesment Plceo 2x1^9 CFU Acute toxicity ssy: 6 dys Norml generl helth sttus: ctivity nd ehvior No difference in ody weight gin or loss No cteril trnsloction (lood, liver or mesenteric lymph nodes) Norml kidney function Norml liver function Dose: 1 times higher thn in humns Immunocompetent mice Immunosuppressed mice Norml pncretic function Norml Gut mucos integrity Norml gut integrity Immunosupression reduced: -gloet cells nd villus height/crypt depth rtio in jejunum. -crypt width in the colon. - CECT 7771 reversed the reduced height/crypt depth rtio in jejunum Antiinflmmtory response CECT 7771 reduced pro-inflmmtory cytokines CECT 7771 restored inflmmtory cytokines of immunosuppressed mice ngoing experiment Chronic toxicity ssy: 3 months Fernández-Murg & Snz, Plosne 215

31 pticl Density (6 nm) Douling time (mins) 6.2 n the wy to evlute new proiotics in vitro functionl chrcteriztion of WBE nd CECT 7771 intervention Preiotic & Proiotic intervention for FD Anti-oesity strtegies Functionl chrcteriztion in vitro in vivo in vitro chrcteriztion of the dietry ptterns nd metolic phenotypes of CECT 7771 Bcteroides spp: glycn enriched environments Cron source (.5% w/v) Glucose inulin Whet Brn Extrct (WBE) gum ric pectin type II mucin,8,6,4,2 Growth fitness of CECT 7771 in different dietry fiers Growth phenotypes Douling time in the log phse * * Glucose Gum ric WBE Inulin Mucin Pectin WBE non-digestile crohydrtes: Arinoxylns rinoxyln-degrding enzymes Arinoxyln oligoscchrides (AXS) 2 CECT Time (hours) Metolic helth SCFA (propionte)

32 6.2 n the wy to evlute new proiotics in vivo functionl chrcteriztion of WBE nd CECT 7771 intervention Preiotic & Proiotic intervention for FD Anti-oesity strtegies Functionl chrcteriztion in vitro in vivo in vivo functionl chrcteriztion on metolic phenotype of WBE+ CECT 7771 intervention C57BL/6 DIET INDUCED BESITY (DI) Kcl (%) Proteins CH Sucrose Ft WBE WHEAT BRAN Kcl (%) PRTEINS CH SUCRSE FAT Kcl (%) PRTEINS CH SUCRSE FAT WHEAT EXTRACT BRAN E Control HFHS HFHS HFHS+F HFHS+WBE Plceo (1% skimmed milk) Plceo (1% skimmed milk) CECT 7771 (1X1 7 CFU) Plceo (1% skimmed milk) -Stndrd Diet HFHS-High Ft High Sucrose diet HFHS+WBE-HFHS+WBE weeks CECT 7771 (1X1 7 CFU) 14 GTT 17 IMMUNMETABLIC PARAMETERS

33 Body weight gin (g) epwat(g)/1g Bw Glycemi (mg/dl) AUC Body weight gin (g) AUC Body weight (g) Glycemi (mg/dl) Body weight ** * ** * ** * 6.2 n the wy to evlute new proiotics in vitro functionl chrcteriztion of WBE nd CECT 7771 intervention p t lest <.5 -Vehicle HFHS week HFHS+WBE HFHS+WBE HFHS ** ** * * ** ** ** * * Preliminry results * Time (min) HFHS C57BL/6 HFHS HFHS+WBE week rl Glucose (2g/Kg) ** 5 * GTT Glycemi t lest p< Vehicle p t lest <.5 HFHS plceo plceo HFHS * week HFHS HFHS+WBE HFHS+WBE HFHS+WBE t lest B. p<.1 uniformis HFHS HFHS+WBE plceo p.8 HFHS HFHS+WBE * HFHS+WBE min week 1 ** * plceo plceo HFHS HFHS+WBE t lest p<.5 HFHS HFHS+WB

34 6.2 n the wy to evlute new proiotics in vitro functionl chrcteriztion of WBE nd CECT 7771 intervention NGING EXPERIMENTS IMMUNMETABLIC PARAMETERS ENERGY HMESTASIS IMMUNE HMESTASIS Hypothlmus SCFA receptors Duodenum Epithelium WAT Ileum Hypotlmic neuropeptides Colon Lmin propri Liver BAT WAT Liver Energy metolism BAT Triglycerides Cholesterol Glucose Leptin Insulin Gut hormones Blood Fecl smples Lympoid cells: B cells, T cells, IEL Myeloid cells: mcrophges, ILCs Cytokines Gut microiot composition

35 THANK YU FR YUR ATTENTIN MARINA RMANÍ-PÉREZ