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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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What are biopharmaceuticals
What Are Biopharmaceuticals? Enteric Bacteria

  • Medical drugs created using biotechnology

  • Include:

    • interferons

    • hormones

    • clotting factors

    • vaccines

    • antibodies


Tuberculosis
Tuberculosis Enteric Bacteria

  • caused by Mycobacterium tuberculosis

  • 8.7 million contracted TB in 2011

  • 630,000 cases of MDR-TB

    • resistant to most powerful tuberculosis drugs



Going further
Going Further Enteric Bacteria


Bibliography
Bibliography Enteric Bacteria

"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/>.



Purpose
Purpose Enteric Bacteria

  • 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
Competing Technologies Enteric Bacteria

  • 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 technologies1
Competing Technologies Enteric Bacteria

  • 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
Design Enteric Bacteria

  • 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 Enteric Bacteria

Signaling

molecule

Repressor

Insulin

production


P Enteric Bacteria

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


Expected results
Expected Results Enteric Bacteria

  • During times of carbohydrate intake  insulin production by IGEBs


Advantages
Advantages Enteric Bacteria

  • Fewer required treatments

  • Completely internal system

  • Self-adjusting system

  • Very flexible lifestyle


Potential problems
Potential Problems Enteric Bacteria

  • Surviving gastrointestinal tract

  • Adhering to villi in small intestine

  • Ensuring adequate absorption of insulin

  • Horizontal gene transfer?


Testing
Testing Enteric Bacteria

  • 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


Bibliography1
Bibliography Enteric Bacteria

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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