A Novel Class of Living Medicines
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1 A Novel Class of Living Medicines Synthetic Biotic TM medicines to perform and deliver critical therapeutic functions to treat diseases throughout the body Genetically Engineered E. coli Nissle attenuates hyperammonemia in a mouse model of hepatic encephalopathy by metabolizing gut ammonia and increasing urea production 1 Yossi Dagon Digestive Disease Week
2 Disclosures Yossi Dagon is an employee of Synlogic Inc. 2
3 Synthetic Biotic Medicines: A Novel Class of Living Medicines Synthetic Engineered bacteria With designed genetic circuits To degrade metabolites that induce disease or synthesize substances to treat disease Biotic: E. coli Nissle as chassis: Widely-used oral probiotic Leverage the safety of probiotic Found within natural human microbiome Amenable to genetic manipulation Synthetic Biology + Bacteria = Synthetic Biotic Medicine Therapeutic delivered locally to treat systemic diseases 3
4 A genetically engineered E. coli Nissle converts Ammonia into Arginine Mechanism of Action: Ammonia / NH 4 Cl Ammonia Arginine Glutamate arge argc argb argh Metabolic Conversions Arginine argr argd argf argl argg cara carb FNR FNR arga fbr Probiotic bacteria: E. coli Nissle Urea cycle Arginine Under normal conditions, urea cycle metabolizes ammonia into urea Urea Where ammonia is not efficiently metabolized via urea cycle, provides an alternative mechanism 4
5 Produces Arginine and Consumes Ammonia Arginine Production by Ammonia Consumption by A r g i n i n e ( m M ) S Y N S Y N B Can orally administrated lower systemic T im e (h ) T im e (h ) A m m o n ia C o n s u m p t i o n ( m M ) S Y N S Y N B hyperammonemia in an animal model of hepatic encephalopathy? 5
6 A thioacetamide-induced hepatic encephalopathy model in BALB/c mice 6
7 lowers blood ammonia levels in TAA-induced liver injury BALB/c mice fed 70% high protein diet Blood ammonia µmol/l * 7 Vehicle CHOW HK 1X10 10 SYN094 1X X10 9 5X10 9 1X10 10 HP (70%) SYN094 unengineered E. coli Nissle HK Heat Killed What is the mechanism by which lowers blood ammonia levels in the TAA-induced liver injury model? * p<0.05 vs. HP (70%) SYN094 1X10 10 HP (70%) 1X10 9 HP (70%) HK-SYN1020 1X10 10 (ttest)
8 and SYN094 improve survival in BALB/c mice following chronic (20 weeks) treatment with TAA Vehicle Percent Survival SYN094 Thioacetamide Vehicle 8 Weeks
9 and SYN094 improve intestinal permeability in BALB/c mice Following chronic treatment with TAA FITC-Dextran TAA-induced liver injury Bacterial products SYN094 Increased intestinal permeability 9 *P<0.05
10 attenuates colon inflammatory cytokines in BALB/c mice Following chronic treatment with TAA qpcr analysis of colon mrna expression: Colon Il-6 Colon TNFα TAA-induced liver injury Bacterial products SYN094 inflammatory markers Il-6 TNFα 10 *P<0.05
11 The effect of on urea blood levels in BALB/c mice Following chronic treatment with TAA Blood urea TAA-induced liver injury ureagenesis Blood urea 11 *P<0.05
12 Does oral administration of affect the development of TAA-induced liver disease? BALB/c mice were co-treated with and TAA for 4 weeks 12
13 and SYN094 improve liver enzymes in a TAA-induced liver injury in BALB/c mice Liver enzyme: AST Liver enzyme: ALT TAA-induced liver injury Liver enzymes SYN * p<0.05 (ttest)
14 lowers blood ammonia in a TAA-induced liver injury in BALB/c mice Blood ammonia * TAA-induced liver injury µmol/l Blood ammonia SYN * p<0.05 (ttest)
15 lowers hepatic inflammatory cytokines expression in TAA-induced liver injury in BALB/c mice Liver Il-6 Liver TNFα TAA-induced liver injury Il-6 15 * p<0.05 (ttest)
16 lowers liver fibrosis markers in TAA-induced liver injury in BALB/c mice Liver TGFβ Liver αsma TAA-induced liver injury Liver fibrosis markers 16 *P<0.05
17 lowers liver fibrosis markers in TAA-induced liver injury in BALB/c mice H&E staining and scoring Microvascular Steatosis (score) Necrosis (score) Apoptosis (score) Control TAA+Vehicle Microvascular Steatosis (score) Necrosis (score) Apoptosis (score) TAA+SYNB102 0 TAA+SYN094 TAA+ Control Vehicle SYN094 Control Vehicle SYN094 Control Vehicle SYN094 H&E-stained sections were assessed by a pathologist blinded to the treatment groups at the time of scoring 17 TAA TAA TAA
18 Summary and Conclusions is a modified E. coli Nissle that consumes ammonia and produces arginine Oral administration of lowers systemic hyperammonemia in a dose dependent manner and elevates blood urea levels in a TAA mouse model of hepatic encephalopathy. and the unengineered strain, SYNB094, improve survival in a chronic TAA mouse model and reduce gut permeability and gutinflammation. partially ameliorates TAA-induced liver injury as demonstrated by lowering elevated liver enzymes, inflammatory cytokines and markers of fibrosis. is currently being evaluated in a phase 1b/2a trial for treatment of hyperammonemia in patients with cirrhosis. 18
19 Acknowledgements Thank you! Hyperammonemia Team: Junning Wang, Lauren Renaud, Mylene Perreault,, Binh Ha, Denise Wong, Charlie Fox, Eugene Antipov, Vincent Isabella, John Thomas, Mike James, Mary Castillo, Caroline Kurtz 19
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