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

Biopharmaceuticals & MDR- TB/XDR-TB

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

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

Synthetic Gene Circuit for Drug Screening

Going Further

Bibliography "Expanding Nature’s Toolkit: How Synthetic Biology Is Changing the Face of Medicine." Sciencebuz. N.p., Web. 8 July Tomilson, Catherine. "Ethionamide." TB Online. Global Tuberculosis Community Advisory Board, 6 Sept Web. 9 July Weber, Wilfried, Ronald Schoenmakers, Bettina Keller, Marc Gitzinger, Thomas Grau, Marie Daoud-El Baba, Peter Sander, and Martin Fussenegger. "A Synthetic Mammalian Gene Circuit Reveals Antituberculosis Compounds." Proceedings of the National Academy of Sciences of the United States of America (2008): Web. 6 July Weber, Wilfried. "Synthetic Biology in Drug Discovery and Combating Drug Resistance." Lecture. Synthetic Biology Workshop - From Science to Governance. Sofitel Hotel, Brussels. 18 Mar Public Health. European Commision. Web. 6 July World Health Organization. "Tuberculosis (TB)." WHO. United Nations, Web. 7 July

Insulin-generating Enteric Bacteria (IGEBs)

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)

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

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

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

Succinate Signaling molecule Repressor Insulin production

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

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

GlucoseInsulin production 00 11

GlucoseInsulin production absent150 units present2000 units

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

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

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

Bibliography Bowen, R. "Absorption of Amino Acids and Peptides." Digestion. Colorado State University, 8 July Web. 1 July Crane, C.W., B.Sc., M.B., M.C. Path., F.R.I.C., and George R. W. N. Luntz, M.R.C.P. "Absorption of Insulin from the Human Small Intestine." Diabetes 17 (1968): Print. "Human Insulin Gene, Complete Cds." National Center for Biotechnology Information. U.S. National Library of Medicine, 12 Feb Web. 10 July "MetaCyc Pathway: Glyxoylate Cycle." MetaCyc. BioCyc Database, 04 Dec Web. 10 July Miller, MB, and BL Bassler. "Quorum Sensing in Bacteria." Annual Review of Microbiology 55 (2001): PubMed.gov. Web. 9 July O'Donnell, Stacy, RN, BS, CDE, and Andrea Penney, RN, CDE. "Insulin Injections vs. Insulin Pump." Diabetes Research, Care, Education & Resources. Joslin Diabetes Center, 11 July Web. 9 July "Part:BBa I TAT Signal+INS_A." Registry of Standard Biological Parts. IGEM, 19 Oct Web. 9 July Shichiri, Motoaki, M.D., Nobuaki Etani, M.D., Ryuzo Kawamori, M.D., Kenkichi Karasaki, M.D., Akira Okada, M.D., Yukio Shigeta, M.D., and Hiroshi Abe, M.D. "Absorption of Insulin from Perfused Rabbit Small Intestine in Vitro." Diabetes 22.6 (1973): Diabetes. American Diabetes Association. Web. 2 July Taqa, ME, JL Semmelhack, and BL Bassler. "The LuxS-dependent Autoinducer AI-2 Controls the Expression of an ABC Transporter That Functions in AI-2 Uptake in Salmonella Typhimurium." Molecular Microbiology 42.3 (2001): PubMed.gov. Web. 9 July