Introduction

1. Towards Intelligent Probiotics Chris Brasseaux, David Golynskiy, Tyler Guinn, SameerSant, MituBhattatiry, NimiBhattatiry, Alfredo Flores Human Practices Scheme • Recent global health concerns have focused greater attention on development assistance measures oriented towards addressing chronic health diseases. Advancements in synthetic biology, such as intelligent probiotics, may serve as important tools to enhance current global health initiatives. They may be engineered to recognize issue-specific signals that elicit issue-specific responses, thus enhancing approaches taken towards achieving the first UN Millennium Development Goal: curbing global malnutrition and hunger. Intelligent probiotics provide potential applications in the grand strategic approach to global health, or the practice of “achieving large ends with limited means.” In context of health-oriented grand strategy outlined by Curry et al., applications of intelligent probiotics can employ the following principles: • I. Start with the goal in mind • UTD iGEM’s end goal was the development of an intelligent probiotic using an E.coli chassis, a bowel symbiont. As a proof of purpose, our application involved mitigating inflammatory bowel disease. • II. Tactics matter • It may prove less resource intensive to improve the efficiency of existing materials (probiotics) at addressing the issue of malnutrition rather than simply channeling more resources into combating the problem. • III. Take an ecological approach • On a microscale, our approach involved two goals: i) engineering wound sensing ability (immunobot) and ii) enabling population control (killbot). Killbots were developed as a biosafety tool to manage unintended consequences associated with using immunobots. On a macroscale, we attempt to illustrate the utility of intelligent probiotics to global health organizations currently grappling with the malnutrition issue. Our system may enhance the effectiveness of current initiatives by making them more cost-effective. • Hence, we believe that grand strategic thinking in synthetic biology will better help connect future developments in biotechnology with their intended beneficiaries. Immunobot Killbot • Sensor • Interface • Immunobot • Taxis • Control Unit Introduction Wound healing involves a complex series of biochemical events that progress in distinct stages: inflammation, proliferation and remodeling. While tissue repair takes place during the proliferative and remodeling stages, chronic wounds resulting from inflammatory bowel disease (IBD) are locked in an inflammatory state and resist the body’s natural ability to repair tissue. According to the Centers for Disease Control, IBD is 1 of the 5 most prevalent gastrointestinal diseases in the United States, accounting for 100,000 hospitalizations and $1.7 billion dollars in health care costs each year. With only a few durable treatments, these disorders remain without a cure. FGFR-ToxRchimeric receptor activates CTX-controlled chemotaxis, thus connecting FGF wound signaling to movement. Fibroblast growth factor (FGF), a wound signal, induces the dimerization of FGF receptor (FGFR), triggering FGF signal transduction. Intracellular levels of CheY-P and CheY control flagellar motors (FliM). High [CheY-P] results in “tumbles” (no net movement) whereas high [CheY] results in “runs” (straight-line movement). Design Approach Probiotics are living microorganisms that can confer a benefit to their host. Live cultures present in regular yogurt products are examples of everyday probiotics. We imagined an intelligent probiotic that can process inputs that indicate tissue damage and produce a programmed output. While the human body itself consists of 100 trillion cells, the bowel hosts 10 times as many microorganisms that exist as part of a mutually beneficial symbiotic relationship. This incredible diversity of symbionts represents a potentially powerful engineering platform for intelligent probiotics that address IBD. In 2009, Stanford developed a two-component probiotic that regulates the activity of Th17 and Treg lymphocytes in order to control tissue inflammation and allow normal wound healing to proceed into the proliferative and remodeling phases. We aimed to build on their work in two ways: i) by engineering a wound sensing ability into E. coli, and ii) by developing a means for the user to control probiotic activity. Accomplishments Construct 2 Construct 3 Construct 1 Delivery/Glucose • Entered information detailing at least one new standard BioBrick (BBa_K569001) in the Registry of Standard Biological Parts. • Demonstrated that at least one new BioBrick (BBa_K569001) of our own design and construction works as expected. We also characterized its operation and entered this information and other documentation on the appropriate Registry page. • Improved the characterization of BBa_K131010, an existing BioBrick, and entered our experience in the appropriate Registry page. • Outlined and detailed a new grand strategic approach to Human Practices in connecting intelligent probiotics with ongoing initiatives in the global health arena to enhance their impact. Time Characterization Modeling and Simulations Immunobot Experiments Activation of Receptor Acknowledgements Dimerization of Receptor Transcription and translation of CheZ • We would like to express our sincerest thanks to the following people for their generous support during our iGEM experience: • To our mentors whose guidance which was invaluable to the success of our project. • To Kristina Ehrhardt, NehaKashyap, and LagnajeetPradhan who provided additional support carrying out experiments and also encoding the wiki and Android apps. • To UT Dallas for providing a facility to carry out our lab experiments. • To Dr. Ruth Silversmith (UNC School of Medicine) and Ms. Britta Herzog (University of Göttigen) for providing us some important constructs. • To the Institute of Biological Engineering for hosting us at the Americas Jamboree. • To iGEM for giving our team this incredible opportunity. Applications Killbot Experiments This work has applications in health and medicine. Our engineered two-component system can be integrated with devices that recruit wound repairing agents at the site of tissue damage. More broadly, intelligent probiotics offer great promise as therapeutic tools for health-related issues such as devices that can process inputs such as nutrient imbalances and affect balance-restoring outputs. 1. Dephosphorylation of CheY Prediction of tumbling activity Population 1 (Immunobot) Population 1 (Immunobot) + Population 2 (Killbot) Population 1 (Immunobot) + more Population 2 (Killbot)