Creating Smart Biomaterials using Engineered Bacteria that Cooperatively Reprogram Mammalian Cells

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NON-TECHNICAL ABSTRACTAn ability to reprogram the function of mammalian cells is critical for exploring the scientific underpinnings of cell behavior as well as improving human health. This effort will result in the development of smart biomaterials and experimental systems made up of microparticles carrying engineered bacteria. These hybrid, living-nonliving biomaterials will be used as a tool to reprogram mammalian cell signaling. One advantage of an extendable synthetic biomaterials approach that uses engineered bacteria to regulate mammalian calcium signaling could be the abrogation of the need to directly alter mammalian cells to reprogram their behavior. Thus, the need for vectors such as viruses or the creation of transgenic animal models could be reduced. For this effort, bacteria-bound microparticles will be developed that can deliver synthetic genetic components to mammalian cells and reprogram calcium signaling. Calcium signaling is a ubiquitous system that effects cells ranging from nerves to the cells that line the gut, and controls both slow and fast cellular processes. Next, these particles will be made into ?smart? biomaterials as their living component ? the bacteria ? will be engineered with an ability to collaborate and collectively determine when to transmit genetic components to mammalian cells. Finally, mathematical modeling and computational simulation will be used to explore calcium signaling dynamics in mammalian cells in order to determine when alterations will cause the most significant changes in cell signaling dynamics. The smart biomaterials will then be used to reprogram this signaling. In addition, this effort will be integrated with the development of multiple teaching modules for a bioengineering summer camp, and the final curriculum will be widely disseminated to the broader scientific and educational community.TECHNICAL ABSTRACTThis effort will result in the creation of smart biomaterials that link to engineered intracellular networks necessary for bacteria to cooperatively invade and then reprogram calcium signaling in mammalian cells. First, a synthetic component will be built to enable engineered bacteria to covalently bond to microparticles to create smart biosensing materials that target specific mammalian cell surface markers. Second, different bacterial lines will be programmed with engineered gene circuits that enable them to cooperatively recognize each other and then activate mammalian cell invasion capabilities. This cooperation further ensures that only specific mammalian cells are targeted. Finally, both the synthetic bonding and the cooperative invasion circuitry will be used to reprogram calcium signaling in mammalian cells. Invading bacteria will use RNA interference to alter the dynamics of calcium signaling by knocking down expression of calcium pathway enzymes. Selection of these target enzymes will be informed by modeling of single cell calcium dynamics. This effort will combine recent advances in engineered cellular invasion, mammalian synthetic biology, cell surface display, and engineered quorum sensing to create bacteria-laden, smart biomaterials that reprogram mammalian cells. The proposed effort includes research to advance discovery and understanding in the field of smart biomaterials while also promoting teaching, training, and learning through the development of a new activities targeted toward high school students.