Transcript Biopharmaceuticals - BLI-Biotech

Synthetic Biology:
Biopharmaceuticals & Insulingenerating Enteric Bacteria
Vi Nguyen
Biopharmaceuticals & MDRTB/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 2011
• 630,000 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., 2012. Web. 8 July 2013.
<http://sciencebuz.com/articles/expanding-nature’s-toolkit-how-synthetic-biology-ischanging-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): 9994998. 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/>.
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
AI-2 signaling molecules
lsr transport
cassette
ABC transporter
ATP
ATP
ADP
ADP
AI-2
P
P
LsrR
LuxS
succinate
prepoinsulin
insulin
ATP
ADP
LsrR
LuxS
lsr promoter TAT peptide
export signal
INS
insulin molecules
Expected Results
• During times of carbohydrate intake 
insulin production by IGEBs
Glucose
0
Insulin production
0
1
1
Glucose
absent
Insulin production
150 units
present
2000 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 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): 16599. 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): 77793. PubMed.gov. Web. 9 July 2013. <http://www.ncbi.nlm.nih.gov/pubmed/11722742>.