CAREER: Multi-Gradient Microtissue Arrays to Analyze Patient-Derived Cancer Cell Modulation by Combinatorial Electrical and Chemical Stimulation

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PI: Verbridge, Scott S.Proposal #: 1652112Cancer is the second leading cause of death in the United States, with some of the poorest results being for highly infiltrative brain tumor glioblastomas (GBMs), for which the 5-year survival rate is less than 10% even with combination treatments of surgery, radiation and chemotherapy (drugs). Surgery and drugs fail to eliminate the malignant cells that infiltrate centimeters beyond the bulk central mass and radiotherapies damage healthy and malignant tissues alike. This project builds on preliminary studies in tissue-engineered models of GBM that demonstrate that pulsed electric fields (PEFs) induce selective destruction of tumor cells by targeting their altered electrical properties, e.g., changes in surface charge, membrane capacitance and ion channel expression. Furthermore, this effect, which targets a physical hallmark of the tumor leaving normal cells relatively unaffected, is enhanced by the addition of drug therapies, which target alterations in tumor cell chemical properties. The goal of this project is to develop a platform to enable efficient screening of the effects of combined physical/drug treatments on patient-derived cells. This novel platform will expand understanding of the beneficial effects of the combined approach and underpin basic science and clinically important advances in a new precision medicine paradigm. The education and outreach plan includes: 1) expanding the "Cancer Engineering" curriculum, outreach, and diversity at Virginia Tech through new and updated courses, research training opportunities for graduate and undergraduate students and supporting K-12 activities for economically disadvantaged students in Southwest Virginia and 2) establishing an "Ask a Virginia Tech Scientist" web column, which will cover a wide spectrum of topics in science, technology, engineering and math (STEM), for local K-12 students and teachers and community engagement. Effective precision cancer medicine protocols will ultimately depend on multi-targeted therapies to treat highly heterogeneous and adaptable tumors such as GBM. The platform developed in this project will be the very first tissue array microchip with an overlay of multiple types of dynamic therapeutic gradients (e.g. electric field amplitude and drug concentration) in orthogonal directions. The platform will be compatible with high-resolution live measurement of cellular dynamics, and coupled to powerful microfluidic molecular analysis to characterize the epigenomic alterations in stimulated cells, uniquely enabling broad-spectrum parametric analysis of combinatorial electrical/molecular treatments of patient-derived cells. The research plan is organized under three objectives: 1) develop a multi-dimensional electrical/chemical gradient microtissue array chip compatible with high-resolution confocal imaging and impedance measurement, 2) develop high-resolution epigenomic analysis with novel coupling to microfluidic chromatin-immunoprecipitation (ChIP), and 3) execute broad-spectrum parametric studies of combinatorial electrical/chemical treatments using patient GBM cells. Successful completion of these objectives will be crucial to: 1) reveal therapeutic synergies possible through a combination of PEFs and drugs, 2) optimize combinatorial treatments with minimal input material (e.g. cells, reagents), and 3) test molecular hypotheses related to treatment synergies. The data collected will provide basic science insights for a new and effective treatment strategy for infiltrative tumors. The combined physical and chemical approaches to targeting both the dense core, as well as the diffuse infiltrative zones will potentially altering the paradigm of GBM treatment.