Glioblastoma (GBM) is the most aggressive tumor of the central nervous system, with a median survival rate of
less than 16 months after diagnosis. Despite extensive research efforts, GBM remains incurable due to its highly
invasive that make complete surgical resection nearly impossible and tumor recurrence inevitable. While it is
recognized that GBMs characteristic invasion is the major determinant of poor clinical outcome, the genetic and
epigenetic regulation of GBM invasion remains poorly understood. To improve GBM treatment options, a
comprehensive understanding of the genetic and epigenetic regulation of GBM invasion along with efficient
targeting modalities are urgently needed. Recent efforts from our lab have included an analysis of human
histology and single-cell RNA sequencing data from ten GBM patients classified as invasive or nodular. Through
these experiments, we identified Crystallin Alpha B (CRYAB) as the top candidate gene upregulated in invasive
GBM samples and saw that overexpression of CRYAB in-vivo increased the invasive phenotype and resulted in
rapid post-operative tumor recurrence. Although these studies suggest that targeting genetic regulators of
invasion may serve as a promising therapeutic for GBM outcome, these analyses were performed by comparing
invasive and noninvasive samples from different patients, and the mechanism by which these invasive genes
drive invasion, including chromatin accessibility that regulates their expression, and how these invasive cells
interact with various cell types of the tumor microenvironment remains unexplored. Therefore, a comprehensive
analysis that elucidates both the genetic and epigenetic regulators of GBM invasion with spatial context in GBM
samples from the same patient are warranted. In Aim 1 I will perform spatial single cell RNA sequencing and
spatial Assay for Transposase-Accessible Chromatin (ATAC) sequencing on GBM tumor samples with matched
pieces from identified contrast enhancing tumor core and non-contrast enhancing invasive tumor. With this
dataset, I will distinguish invasive tumor cells from cells in the tumor microenvironment (TME) and noninvasive
tumor bulk, and use this data to identify novel genetic and epigenetic regulators of GBM invasion. In Aim 2, I will
explore the mechanism and evaluate the in-vitro and in-vivo impact targeting i) previously identified invasive
gene CRYAB, and ii) candidate gene identified in Aim 1, has on the invasive phenotype by leveraging our lab’s
expertise in nanoparticle design and synthesis for delivery of genetic materials. This research will reveal critical
insight into the mechanisms underlying GBM invasion and progression, and may also provide candidates to
serve as novel prognostic and therapeutic candidates.