Research Testbed 2

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Abstract Brain tumors remain among the most malignant and deadly cancers, with adult glioblastoma (GBM) having a median survival of 15 months and five-year survival of less than 10 percent (Stupp et al., 2009), despite surgery, chemotherapy (temozolomide), and radiation. While each of these approaches provides an overall survival benefit of a few months, they do not offer a longer-term survival benefit or cure, even in combination. Thus, new therapy approaches are sorely needed. To rationally develop therapies, it is critical that we have better experimental systems for both fundamental and preclinical investigations that faithfully recapitulate key features of the human disease while bringing the full power of state-of-the-art engineering approaches to bear including modeling and simulation. In unpublished work, we have found that the Sleeping Beauty (SB) transposase system we developed can be used to produce two major GBM, proneural (PN) and mesenchymal (MES), in immune- competent wild-type mice using known human oncogenic drivers of glioblastoma, and that individual cancer cells can be tracked in live tumors via multichannel fluorescence imaging. We hypothesize that mechanical forces mediate a key targetable difference between PN and MES subtypes, and that the immune cold/hot signatures of PN and MES subtypes represents a second key targetable difference between subtypes. To test these hypotheses, we will measure cellular force and signaling dynamics of cancer cells and T cells in live GBM tumors (Aim 1) and Measure immune cell dynamics in live GBM tumors (Aim 2). In particular we will measure cell traction forces, kinase and GEF signaling, and extracellular matrix architecture in live tumors and brain tissue. In addition, we will track antitumoral T cells, and protumoral microglia and bone marrow-derived macrophages, and their correlations with each other and GBM cells, in the SB mouse models of PN and MES. These experiments will be used to parameterize the Cell Migration Simulator (CMS) and Brownian Dynamics Tumor Simulator (BDTS) to create predictive models that are GBM subtype-specific. In most cases, these measurements will be the first of their kind in live GBM tumors, and will inform cell and tumor scale biophysical models. In so doing, RTB2 will be providing a state-of-the-art integrated experimental-computational testbed for development of imaging modalities in the TECH unit.