Stony Brook iGEM Nephrotides: Regulating Blood Glucose Levels With - - PowerPoint PPT Presentation

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation,

• 2015. Web. 23 Sept. 2015.

Stony Brook iGEM Nephrotides:

Regulating Blood Glucose Levels With Peptide Secreting Microbes

Antibiotic Resistance

Source: econlife.com

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation,

• 2015. Web. 23 Sept. 2015.

Melittin

Source: "Italian Bee."

• Wikipedia. Wikimedia

Foundation, n.d. Web. 21

• Sept. 2015.

Source: SBU iGEM 2014

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation,

• 2015. Web. 23 Sept. 2015.

Melittin Optimization

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation,

• 2015. Web. 23 Sept. 2015.

Infectious Disease vs. Chronic Disease in the US

*Compiled from data conducted by David S. Jones, M.D., Ph.D., Scott

• H. Podolsky, M.D., and Jeremy A. Greene, M.D., Ph.D. at Harvard

Medical School in The Burden of Disease and the Changing Task of Medicine.

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation, 2015. Web. 23 Sept. 2015.

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation,

• 2015. Web. 23 Sept. 2015.

Type II Diabetes

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation,

• 2015. Web. 23 Sept. 2015.

Sodium Glucose Transporter 1 and 2

Lumen Blood Na+ Glucose SGLT1 Na+ SGLT2 GLUT2 K+ Na/K ATPase Proximal Tubule

Based on: Wilding, John P.H. "SGLT2 Inhibitors for the Treatment of Diabetes." MedScape Multispecialty. Medscape, LLC, 3 July 2009. Web. 23 Sept. 2015.

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation,

• 2015. Web. 23 Sept. 2015.

Glutamine-Serine-Proline (QSP)

AMG uptake [% of control]

without tripeptides with QCPac*

Source: Vernaleken, A., et. al.”Tripeptides…” QSP has the same functionality as QCP.

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation,

• 2015. Web. 23 Sept. 2015.

Project Overview

EnvZ Proline Iminopeptidase OmpA HlyA System Oligopeptides 1 2 Active Tripeptide Solute Cleaver Cell Sensor Cell

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation,

• 2015. Web. 23 Sept. 2015.

The Sensor

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation,

• 2015. Web. 23 Sept. 2015.

The Secretion System

Source: Pbase.com Dalbey, Ross E., and Andreas Kuhn. "Protein Traffic in Gram- negative Bacteria – How Exported and Secreted Proteins Find Their Way." Oxford University Press (2012): n. pag. 1 Nov.

• 2012. Web. 23 Sept. 2015.

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation,

• 2015. Web. 23 Sept. 2015.

Construct Design

Oligopeptides Membrane Secretion

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation,

• 2015. Web. 23 Sept. 2015.

Peptide Production

Ladder Peptides 1:2 Negative Control Peptides 1:5 Peptides 1:10 Negative Control OmpC Antibiotic Resistance

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation,

• 2015. Web. 23 Sept. 2015.

The Protease

Proline Iminopeptidase

Source: European Protein Data Base

Proline Endopeptidase

Source: Protein Data Base

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation,

• 2015. Web. 23 Sept. 2015.

Protease Construction

Proline Iminopeptidase Membrane Displayed Proline Iminopeptidase

Sensor Design (cAMP Sensor)

Sensor Design (Insulin-SLN1 Chimera)

I-TASSER Generated Chimera Model

• Successful fusion of various tyrosine kinase receptor

ligand binding domains (Pacifi, et. al).

• Successful fusion of Human Epidermal Growth Factor

Receptor ligand binding region with intracellular region of SLN1 (Ostrander, et. al).

Community Involvement

Collaboration: University of Georgia Atlantic iGEM team

Outreach

Educating the Future Generation

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation,

• 2015. Web. 23 Sept. 2015.

Human Practices Mind Map

Fear genetically engineered

• rganisms

Under informed of synthetic biology techniques Skeptical of using E.coli to treat disease

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation,

• 2015. Web. 23 Sept. 2015.

Human Practices

Harlod Carlson, MD Chairman of IRB Human Practices Institutional Review Board

Source:"Biblioguide En Thérapie Génique." Ecole Polytechnique Federale De Lausanne. EPFL, n.d. Web. 14 Sept. 2015.

Further Research

• Test the system in vitro
• Test the system in vivo in mice

In Summary

• Our ¡system ¡is ¡an ¡insulin ¡independent ¡treatment ¡for ¡type ¡two ¡diabe4c ¡

pa4ents ¡

• The ¡glucose ¡sensor ¡creates ¡a ¡responsive ¡medicine ¡produc4on ¡system ¡

in ¡vivo ¡

• Encapsula4on ¡of ¡engineered ¡cells ¡allows ¡for ¡a ¡contained ¡and ¡safe ¡

delivery ¡system ¡in ¡conjunc4on ¡with ¡pa4ent ¡needs ¡

"Incidence and Prevalence of Chronic Disease." (MPKB). Autoimmunity Research Foundation,

• 2015. Web. 23 Sept. 2015.

Acknowledgements

Faculty Advisers

John Peter Gergen, Ph.D. Gabor Balazsi, Ph.D. Kevin Czaplinski, Ph.D. Ete Chan, Ph.D. Joshua Rest, Ph.D. Yizhi Meng, Ph.D.

Undergraduate Advisers

Janki Patel Nicolai Alexandre Aquino Tayco Gurkamal Kaur Helen Liu

Special Thanks To

The following departments of Stony Brook University:

• Department of Undergraduate Biology
• Department of Biochemistry and Cell

Biology

• Department of Biomedical Engineering
• Department of Evolution and Ecology
• Department of Pharmacological Sciences
• Department of Pathology
• Office of Scientific Affairs
• Stony Brook Laufer Center
• Undergraduate Research & Creative

Activities Harold Carlson, M.D. Steven Glynn, Ph.D. Wali Karzai, Ph.D. David Thanassi, Ph.D. Jessica Seeliger, Ph.D. Markus Seeliger, Ph.D. Marvin O'Neal, Ph.D.