TYPE 2 DIABETES MELLITUS AFFECTS OVER 8% OF AMERICAN ADULTS. THE FIRST-LINE ORAL TREATMENT FOR TYPE 2 DIABETES IS METFORMIN, A DRUG THAT LOWERS PLASMA GLUCOSE AND IMPROVES METABOLISM. HOWEVER, METFORMIN AND OTHER ANTI-DIABETIC MEDICATIONS HAVE A HIGH RATE OF FAILURE OVER TIME, SO TYPE 2 DIABETICS COMMONLY NEED TO TAKE MORE THAN ONE MEDICATION AND EVENTUALLY PROGRESS TO MORE INTENSIVE TREATMENT SUCH AS INSULIN THERAPY. THEREFORE, ACCESSIBLE AND NUTRITIOUS SUPPLEMENTS ARE NEEDED THAT ARE DELIVERED IN CONJUNCTION WITH METFORMIN TO IMPROVE DIABETIC CONTROL. MANY TYPE 2 DIABETICS MEET THE CRITERIA FOR METABOLIC SYNDROME, A DISEASE THAT IS CHARACTERIZED BY HIGH BLOOD GLUCOSE, HIGH BLOOD PRESSURE, DYSREGULATED LIPID METABOLISM, AND CENTRAL OBESITY. METABOLIC SYNDROME AND TYPE 2 DIABETES ARE CHARACTERIZED BY CHRONIC, LOW-GRADE INFLAMMATION AND GUT MICROBIAL DYSBIOSIS. GUT MICROBIAL DYSBIOSIS REFERS TO A GUT MICROBIOME (THE COMMUNITY OF BACTERIA THAT LIVE IN THE INTESTINE) THAT HAS BECOME DYSREGULATED AND CAN CAUSE DISEASE. THE ASSOCIATIONS BETWEEN METABOLIC SYNDROME AND GUT MICROBIAL DYSBIOSIS HAVE BEEN EXTENSIVELY STUDIED AND CORRECTING GUT DYSBIOSIS IS PROPOSED AS A POTENTIAL THERAPY FOR METABOLIC SYNDROME AND TYPE 2 DIABETES.METFORMIN IS KNOWN TO IMPROVE OBESITY-ASSOCIATED INFLAMMATION AND ALTERS THE GUT MICROBIOME COMPOSITION, WHICH IS THOUGHT TO IMPROVE GUT DYSBIOSIS AND LOWER SYSTEMIC INFLAMMATION. PROBIOTICS ARE OF GREAT INTEREST AS THEY ARE ALSO THOUGHT TO MODULATE THE GUT MICROBIOME AND IMPROVE INFLAMMATION. SOME PROBIOTICS HAVE ALSO BEEN SHOWN TO INTERACT WITH THE IMMUNE SYSTEM. OUR PAST RESEARCH HAS SHOWN THAT A STRAIN OF PROBIOTICS CALLED BIFIDOBACTERIUM ANIMALIS SPP. LACTIS (BB-12) IN A YOGURT SMOOTHIE LOWERS TWO INFLAMMATORY MARKERS IN HEALTHY ADULTS, SUGGESTING THAT PROBIOTICS MAY EXERT AN ANTI-INFLAMMATORY EFFECT. FURTHERMORE, OUR PREVIOUS RESEARCH HAS SHOWN THAT THE METHOD OF PROBIOTIC DELIVERY AFFECTS ITS EFFICACY, SUGGESTING THAT ADDING PROBIOTICS TO A DAILY YOGURT SMOOTHIE MAY BE A MORE EFFECTIVE DELIVERY METHOD. THE POTENTIAL BENEFICIAL ACTIONS OF BOTH METFORMIN AND PROBIOTICS HAVE LED TO THE HYPOTHESIS THAT THEY COULD ACT IN COMBINATION TO ALLEVIATE INFLAMMATION; HOWEVER, THIS HAS NOT BEEN TESTED IN A RANDOMIZED CONTROLLED TRIAL OF TYPE 2 DIABETES. THE PROPOSED PROJECT ADDRESSES THIS KNOWLEDGE GAP BY SYNTHESIZING RESULTS FROM PUBLISHED RESEARCH TO EXPLORE THE RELATIONSHIP BETWEEN METFORMIN AND PROBIOTIC USE ON MICROBIAL POPULATIONS WITH METABOLIC SYNDROME AND TYPE 2 DIABETES; ADDING AN ADDITIONAL ENDPOINT TO AN EXISTING CLINICAL TRIAL TO EXAMINE THE EFFECT OF ADDING A DAILY PROBIOTIC YOGURT TO EXISTING METFORMIN TREATMENT ON INFLAMMATORY AND MICROBIOME OUTCOMES; AND CONDUCTING A MECHANISTIC STUDY TO EVALUATE THE CAUSAL ROLE OF THE GUT MICROBIOME IN MEDIATING THE BENEFICIAL EFFECT OF METFORMIN AND PROBIOTICS ON SYSTEMIC INFLAMMATION. THE OVERARCHING HYPOTHESIS IS THAT PROBIOTIC BB-12 IN A YOGURT SMOOTHIE ADDED TO METFORMIN TREATMENTWILL MODULATE THE GUT MICROBIOME AND IMPROVE INFLAMMATION IN TYPE 2 DIABETES.THIS PROJECT INCLUDES A META-ANALYSIS OF PUBLISHED LITERATURE, A SUBGROUP ANALYSIS OF AN ONGOING CLINICAL TRIAL, AND A MECHANISTIC MOUSE STUDY. IN THE META-ANALYSIS, ACCESSIBLE DATA WILL BE RE-ANALYZED TO DETERMINE IF METFORMIN AND PROBIOTICS INTERACT WITH SIMILAR GUT BACTERIA. IN THE CLINICAL TRIAL, TYPE 2 DIABETICS WITH METFORMIN TREATMENT WILL BE RANDOMIZED TO RECEIVE EITHER A DAILY PLACEBO YOGURT SMOOTHIE OR A DAILY YOGURT SMOOTHIE WITH BB-12 PROBIOTICS ADDED. THIS STUDY WILL DIRECTLY TEST IF ADDING PROBIOTIC YOGURT TO METFORMIN TREATMENT LOWERS CIRCULATING INFLAMMATORY MARKERS. THE MECHANISTIC MOUSE STUDY WILL EVALUATE THE CAUSAL ROLE OF THE GUT MICROBIOME IN MEDIATING THE ANTI-INFLAMMATORY EFFECT OF METFORMIN AND PROBIOTICS BY TREATING MICE WITH ANTIBIOTICS TO REMOVE THE HOST MICROBIOME AND THEN TREATING MICE WITH METFORMIN AND PROBIOTICS. THIS STUDY WILL DETERMINE IF THE HOST MICROBIOME MEDIATES SOME ANTI-INFLAMMATORY EFFECTS OR IF THERE MAY BE OTHER MECHANISMS THAT ARE INVOLVED.THE OUTCOMES OF THIS STUDY WILL BE DISSEMINATED TO THE SCIENTIFIC COMMUNITY AND THE REGIONAL PENNSYLVANIAN POPULATION OF TYPE 2 DIABETICS WHO MAY BENEFIT FROM THE RESULTS OF THIS STUDY. PEER-REVIEWED MANUSCRIPTS WILL BE SUBMITTED TO SCIENTIFIC JOURNALS AND THE RESULTS WILL BE PRESENTED AT SCIENTIFIC CONFERENCES TO COMMUNICATE THE STUDY RESULTS TO OTHER SCIENTISTS AROUND THE WORLD. LOCALLY, A WORKSHOP WILL BE TAUGHT AT THE PENN STATE MICROBIOME CENTER TO TEACH OTHER FACULTY AND STUDENTS HOW TO ACCESS DATA USED IN THE META-ANALYSIS IN AIM 1. A PENN STATE EXTENSION BULLETIN WILL BE PUBLISHED IN THE DINING WITH DIABETES PROGRAM THAT REACHES TYPE 2 DIABETICS AROUND THE STATE OF PENNSYLVANIA. OVERALL, THIS PROJECT WILL RESULT IN NEW KNOWLEDGE CONTRIBUTING TO IDENTIFYING ACCESSIBLE, NUTRITIOUS DAIRY-BASED SUPPLEMENTS THAT ARE ADDED TO METFORMIN TREATMENT TO FURTHER IMPROVE DIABETIC CONTROL IN THE AIM OF REDUCING THE NEED FOR COMBINATION TREATMENT AND INSULIN THERAPY.