Identification of Secretion Factors that Restructure Neural Tissue in a Model of Low Grade Glioma

Pediatric Low-grade Gliomas account for up to ⅔ of all pediatric tumors. Despite their low grade classification, they have the ability to mutate farther to a grade 3 or 4 neurological tumor, exponentially decreasing treatment and survival rates. Identifying the specific secretion factors associated with these tumors is extremely beneficial towards possible treatment options. CRISPR/Cas9 constructs were implemented to knock out GDF-15 and CCl-3 secretion factors in the presence of a BRAFV600E mutation before concentrated DNA plasmids were injected and integrated into the brains of mice embryos through In Utero Electroporation (IUE). The brains were subsequently stained through an immunohistochemistry procedure with the goal of highlighting the microvascular network and microglial activation in the transfected portions of the sectioned brains. Preliminarily, we were able to identify the increase in microglial activation particularly with the CCL-3 with the BRAFV600E mutation condition as well as an increase to the density of the microvascular network, particularly in the larger cranial blood vessels, in the GDF-15 with the BRAFV600E mutation condition. Further quantitative analysis is required before more concrete conclusions can be made