Increasing barrier function with multispecies probiotics
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1 Increasing barrier function with multispecies probiotics Elsbeth Pekelharing, MSc Science Liaison We strive for the most effective microbiome management solutions with our evidence-based & indication-specific multispecies probiotic formulations PAGE 2 4 th Microbiome R&D and Business Collaboration Forum USA
2 Winclove s Company DNA 60+ B2B customers 30+ countries 70+ formulations 25 years dedicated to DEVELOPMENT, research and manufacturing of PROBIOTICS INDICATION SPECIFIC multispecies probiotics 100+ strains 10 prebiotics DIETARY supplements, FSMP & MD PhDs +10% 100 million daily dosages sold in 2015 PAGE 3 4 th Microbiome R&D and Business Collaboration Forum USA Winclove Research Our Focus Areas 50 RESEARCH PROJECTS started, ongoing, or finished in % of research in 2015 were in HUMANS SPORT TRAVEL >40 External research partners worldwide IBD WOMEN S HEALTH ENT GUT BRAIN AXIS CONSTIP. ANTIBIOTIC ASSOCIATED SYMPTOMS ALLERGIES >50 peer reviewed PUBLICATIONS over the last 10 years 10 MANUSCRIPTS IN THE RESEARCH PIPELINE for publication in 2016 PAGE 4 4 th Microbiome R&D and Business Collaboration Forum USA
3 External research partners PAGE 5 4 th Microbiome R&D and Business Collaboration Forum USA Evidence based formulations: Ecologic PAGE 6 4 th Microbiome R&D and Business Collaboration Forum USA
4 Two key principles I Charactarisation and selection on strain level Lactobacillus rhamnosus X Lactobacillus rhamnosus Y II It is not merely about cellcount PAGE 7 4 th Microbiome R&D and Business Collaboration Forum USA Our view on the mode of action 1 Microbe Microbe Producing antimicrobial agents against pathogens. Compete for colonization space and receptors. Communicate with commensals (e.g. quorum sensing). Modify metabolic environment (e.g. SCFA production) 2 Microbe Epithelial barrier Improve barrier function (e.g. tight junctions) Induce cytoprotective compounds (e.g. defensins). Stimulate mucus production. 3 Microbe Systemic Educate immune system. Enhance signaling response (e.g. Ig and cytokines) Induce immune cell migration (e.g. T and B cells) Moderate responsiveness (e.g. to food antigens) PAGE 8 4 th Microbiome R&D and Business Collaboration Forum USA
5 Epithelial barrier Largest bodily surface that interacts with the outside world: appr 300m 2 PAGE 9 4 th Microbiome R&D and Business Collaboration Forum USA Epithelial barrier Healthy Disturbed situation Thick mucus layer Strong tight junctions Epithelial cells in good condition e.g. LPS in circulation Proinflammatory cytokines Low grade inflammation Associated with e.g. IBD, metabolic syndrome, diabetes, artherosclerose, rheumatoid arthritis, chronic fatigue syndrome, depression, autism, migraine. PAGE 10 4 th Microbiome R&D and Business Collaboration Forum USA
6 Restoring tight junction function 0 Acute pancreatitis Multispecies probiotic - - placebo + (pretreatment) Lutgendorff et al (2009) PAGE 11 4 th Microbiome R&D and Business Collaboration Forum USA How to select the right team? PAGE 12 4 th Microbiome R&D and Business Collaboration Forum USA
7 Selection criteria Stimulate regulatory response Inhibit pro-inflammatory response Barrier function 1 IL Mast cell LPS 4 TEER IL-10 production Inhibition mast cell activation LPS breakdown Trans epithelial resistance PAGE 13 4 th Microbiome R&D and Business Collaboration Forum USA Transepithelial electric resistance bacteria Caco-2 Transwell system Epithelial layer stressed with TNF & IL-1 Assay per individual probiotic strain PAGE 14 4 th Microbiome R&D and Business Collaboration Forum USA
8 Effect of strains on barrier function controle stressor W64-Bac. coagulans W53-B. animalis W23-B. bifidum W28-B. bifidum W119-B. bifidum W25-B. breve W125-B. infantis W51-B. lactis W52-B. lactis W108-B. longum W54-E. faecium W74-L. acidophilus W22-L. acidophilus W37-L. acidophilus W55-L. acidophilus W70-L. acidophilus W63-L. brevis W78-L. brevis W20-L. casei W56-L. casei W79-L. casei W129-L. fermentum W21-L. plantarum W62-L. plantarum W71-L. rhamnosus W24-L. salivarius W57-L. salivarius % TEER W19-Lc. lactis W58-Lc. lactis W122-S. salivarius Stressor (TNF-α + strain) PAGE 15 4 th Microbiome R&D and Business Collaboration Forum USA Formulation Eight bacterial strains Bifidobacterium bifidum W23 Bifidobacterium lactis W52 Lactobacillus acidophilus W37 Lactobacillus brevis W63 Lactobacillus casei W56 Lactobacillus salivarius W24 Lactococcus lactis W19 Lactococcus lactis W58 Concentration: 2,5 billion cfu/gram Corn starch, maltodextrine, vegetable protein, mineral mix (potassium magnesium manganese sulfate) PAGE 16 4 th Microbiome R&D and Business Collaboration Forum USA
9 In vivo studies Barrier function Proof of principle (ongoing) Gut brain general RCT with fmri (ongoing) Depression Animal model RCT pilot (Steenbergen et al, 2015) RCT in elderly (Leiden University, ongoing) RCT in depressive patient (ongoing) follow up planned with antidepressant Migraine Open label pilot (Roos et al, 2015) RCT (analyzing) Diabetes DMT2 RCT (ongoing) II DMT1 fundamental (orientation) Liver cirrhosis RCT (Horvath et al, 2016) PAGE 17 4 th Microbiome R&D and Business Collaboration Forum USA Barrier function in liver cirrhosis Impaired barrier function and prone to bacterial infections Normally neutrophils produce reactive oxygen species (ROS) in response to LPS for microbial clearance Hypothesis: Chronic overstimulation exhaustion Clinical trails show probiotics to improve liver function in different stages of alcoholic fatty liver disease (Horvath et al, 2016) PAGE 18 4 th Microbiome R&D and Business Collaboration Forum USA
10 Ecologic Barrier in Liver cirrhosis Methods Randomized, double blind, placebo controlled study N=92 stable cirrhosis patients (and 51 healthy controls) 6 months intervention, 6 months follow up (Horvath et al, 2016) PAGE 19 4 th Microbiome R&D and Business Collaboration Forum USA Ecologic Barrier in Liver cirrhosis Results No significant improvement on neutrophil phagocytosis, endotoxin load, gut permeability, or inflammatory markers. Improved antimicrobial activity Oxidative resting burst of neutrophils increased significantly Serum neopterin levels were significantly increased (macrophage activity) Liver function improved in probiotic group (Child Pugh score) More dropouts in placebo group PAGE 20 4 th Microbiome R&D and Business Collaboration Forum USA
11 Barrierfunction in gut-brain-axis Breaking down the barriers: the gutmicrobiome, intestinal permeability and stress-related psychiatric disorders. Kelly et al (2015) PAGE 21 4 th Microbiome R&D and Business Collaboration Forum USA Depression rodent study Aarhus University, Denmark Placebo controlled study with 40 healthy rats floating Multispecies probiotic 8 weeks Ecologic Barrier (5x10 9 cfu/day) Forced swim test (7 min) PAGE 22 4 th Microbiome R&D and Business Collaboration Forum USA
12 PAGE 23 4 th Microbiome R&D and Business Collaboration Forum USA MOVIE: Depression human study Leiden University, the Netherlands Randomized, triple blind, placebo-controlled study 40 healthy students Multispecies probiotic (Ecologic Barrier) or placebo 4 weeks, 2 gram daily Questionnaire: Leiden Index of Depression Sensitivity, Revised PAGE 24 4 th Microbiome R&D and Business Collaboration Forum USA
13 Depression human study * * * Cognitive reactivity to sad mood PAGE 25 4 th Microbiome R&D and Business Collaboration Forum USA Publications PAGE 26 4 th Microbiome R&D and Business Collaboration Forum USA
14 Thank you for your attention Any questions? 14
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