DEVELOPING AN ENVIRONMENTAL STIMULI-INDUCED SYSTEM FOR PROTEIN PROXIMITY
Larisa Breden, Guihua Zeng, Roushu Zhang, Wei Wang, Fu-sen Liang.
The University of New Mexico, Albuquerque, NM.
Elevated levels of Fe2+ and H2O2 have been marked in neurodegenerative brains of both Alzheimer’s disease (AD) and Parkinson’s disease (PD) patients, contributing to the pathogenesis. We hypothesize that a novel, synthetic, biology-based gene therapy strategy that incorporates environment-sensing and bio-computing logics (i.e., “AND” Boolean logic gate) can ensure a desired response is generated only when a specific environment exists. This system is based on 2 reported chemically induced proximity (CIP) systems: absicsic acid (ABA) and gibberellin (GA). The systems use exogenous chemical inducers to regulate biological processes via protein proximity. We chemically caged ABA and GA to be used as sensors for Fe2+ and H2O2 and as inducers for respective downstream cellular processes. We predict that when free ABA and/or GA is produced in the presence of Fe2+ and H2O2, the proteins will fuse to a DNA binding or transactivation domain. For the model system, this induces expression of a downstream protein, green florescence protein (GFP), which can later be replaced by a therapeutic protein. We plan to expose the engineered cells to concentrations of Fe2+ and H2O2, monitoring GFP expression with florescence microscopy. The detection of GFP fluorescence only in cells with both Fe2+ and H2O2 concentrations seen in AD and PD will indicate a condition-specific CIP system. By engineering an environment-detecting cell, we address a major issue of cell therapy development. We predict that our system can be modified to respond to the specific biological conditions of many diseases in addition to AD and PD.