1. Synthetic Biology: Biopharmaceuticals & Insulin- generating Enteric Bacteria Vi Nguyen

2. Biopharmaceuticals & MDR- TB/XDR-TB

3. What Are Biopharmaceuticals? Medical drugs created using biotechnology Include: –interferons –hormones –clotting factors –vaccines –antibodies

4. Tuberculosis caused by Mycobacterium tuberculosis 8.7 million contracted TB in 2011 310,000 cases of MDR-TB –resistant to most powerful tuberculosis drugs

5. Synthetic Gene Circuit for Drug Screening

7. Going Further

8. Insulin-generating Enteric Bacteria (IGEBs)

9. Purpose Provide a more convenient means of insulin therapy for diabetics Modify native gut flora to produce insulin (E. coli) Bacteria that produce insulin at ideal times (during glucose intake)

11. Competing Technologies Insulin injections –Pros: Relatively inexpensive Relatively simple to administer –Cons: Requires daily injections Blood glucose spikes Insulin resistance may occur in repeated needle stick areas

12. Competing Technologies Insulin pumps –Pros: More accurate doses Fewer blood glucose spikes More flexible lifestyle Fewer needlesticks –Cons: Expensive Bulky system constantly attached to body Requires extensive training to use

13. Design Determining when to produce large amounts of insulin No glucose  no insulin –Glyoxylate cycle in absence of glucose Modified quorum sensing –Produce large amounts of insulin at certain times

15. Succinate Signaling molecule Repressor Insulin production

16. LuxS lsr promoterLuxSTAT peptide export signal INS succinate AI-2 signaling molecules ABC transporter ATPADP ATP ADP lsr transport cassette LsrR P P TAT export prepoinsulin insulin molecules AI-2 insulin ATP ADP

17. Expected Results During times of carbohydrate intake  insulin production by IGEBs

18. GlucoseInsulin production 00 11

19. GlucoseInsulin production absent150 units present2000 units

20. Advantages Fewer required treatments Completely internal system Self-adjusting system Very flexible lifestyle

21. Potential Problems Surviving gastrointestinal tract Adhering to villi in small intestine Ensuring adequate absorption of insulin Horizontal gene transfer?

22. Testing Insulin production in absence/presence of glucose in environment –Cells exposed to various cycles of glucose absence and presence –Insulin production measured and tracked over time

23. Sources http://ec.europa.eu/health/dialogue_collaboration/docs/ev_2010031 8_co10.pdfhttp://ec.europa.eu/health/dialogue_collaboration/docs/ev_2010031 8_co10.pdf http://sciencebuz.com/articles/expanding-nature%E2%80%99s- toolkit-how-synthetic-biology-is-changing-the-face-of-medicine/http://sciencebuz.com/articles/expanding-nature%E2%80%99s- toolkit-how-synthetic-biology-is-changing-the-face-of-medicine/ http://www.pnas.org/content/105/29/9994 http://www.who.int/topics/tuberculosis/en/ http://www.vivo.colostate.edu/hbooks/pathphys/digestion/smallgut/a bsorb_aacids.htmlhttp://www.vivo.colostate.edu/hbooks/pathphys/digestion/smallgut/a bsorb_aacids.html http://diabetes.diabetesjournals.org/content/22/6/459 http://www.joslin.org/info/insulin_injections_vs_insulin_pump.html http://www.biocyc.org/META/NEW- IMAGE?type=PATHWAY&object=GLYOXYLATE-BYPASShttp://www.biocyc.org/META/NEW- IMAGE?type=PATHWAY&object=GLYOXYLATE-BYPASS http://www.who.int/tb/publications/factsheet_global.pdf