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Synthetic Biology: Biopharmaceuticals & Insulin-generating Enteric Bacteria

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.

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Synthetic Biology: Biopharmaceuticals & Insulin-generating Enteric Bacteria

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  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 • 630,000 cases of MDR-TB • resistant to most powerful tuberculosis drugs

  5. Synthetic Gene Circuit for Drug Screening

  6. Going Further

  7. Bibliography "Expanding Nature’s Toolkit: How Synthetic Biology Is Changing the Face of Medicine." Sciencebuz. N.p., 2012. Web. 8 July 2013. <http://sciencebuz.com/articles/expanding-nature’s-toolkit-how-synthetic-biology-is-changing-the-face-of-medicine/>. Tomilson, Catherine. "Ethionamide." TB Online. Global Tuberculosis Community Advisory Board, 6 Sept. 2012. Web. 9 July 2013. <http://www.tbonline.info/posts/2011/8/24/ethionamide/>. 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 105.29 (2008): 9994-998. Web. 6 July 2013. <http://www.pnas.org/content/105/29/9994>. Weber, Wilfried. "Synthetic Biology in Drug Discovery and Combating Drug Resistance." Lecture. Synthetic Biology Workshop - From Science to Governance. Sofitel Hotel, Brussels. 18 Mar. 2010. Public Health. European Commision. Web. 6 July 2013. <http://ec.europa.eu/health/dialogue_collaboration/docs/ev_20100318_co10.pdf>. World Health Organization. "Tuberculosis (TB)." WHO. United Nations, 2013. Web. 7 July 2013. <http://www.who.int/topics/tuberculosis/en/>.

  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)

  10. 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

  11. 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

  12. 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

  13. Succinate Signaling molecule Repressor Insulin production

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

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

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

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

  18. 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

  19. Bibliography Bowen, R. "Absorption of Amino Acids and Peptides." Digestion. Colorado State University, 8 July 2006. Web. 1 July 2013. <http://www.vivo.colostate.edu/hbooks/pathphys/digestion/smallgut/absorb_aacids.html>. 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): 625-27. Print. "Human Insulin Gene, Complete Cds." National Center for Biotechnology Information. U.S. National Library of Medicine, 12 Feb. 2001. Web. 10 July 2013. <http://www.ncbi.nlm.nih.gov/nuccore/J00265.1>. "MetaCyc Pathway: Glyxoylate Cycle." MetaCyc. BioCyc Database, 04 Dec. 2007. Web. 10 July 2013. <http://www.biocyc.org/META/NEW-IMAGE?type=PATHWAY&object=GLYOXYLATE-BYPASS>. Miller, MB, and BL Bassler. "Quorum Sensing in Bacteria." Annual Review of Microbiology 55 (2001): 165-99. PubMed.gov. Web. 9 July 2013. <http://www.ncbi.nlm.nih.gov/pubmed/11544353>. 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 2013. Web. 9 July 2013. <http://www.joslin.org/info/insulin_injections_vs_insulin_pump.html>. "Part:BBa I761002 TAT Signal+INS_A." Registry of Standard Biological Parts. IGEM, 19 Oct. 2007. Web. 9 July 2013. <http://parts.igem.org/Part:BBa_I761002>. 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): 459-65. Diabetes. American Diabetes Association. Web. 2 July 2013. <http://diabetes.diabetesjournals.org/content/22/6/459>. 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): 777-93. PubMed.gov. Web. 9 July 2013. <http://www.ncbi.nlm.nih.gov/pubmed/11722742>.

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