LC-Orbitrap\nHRMS-Based Proteomics Reveals Novel Mitochondrial\nDynamics Regulatory Proteins Associated with <i>Ras</i><i>V12-</i>Induced Glioblastoma (GBM) of <i>Drosophila</i>

Glioblastoma multiforme (GBM) is the most prevalent and\naggressive\nbrain tumor found in adult humans with a poor prognosis and average\nsurvival of 14–15 months. In order to have a comprehensive\nunderstanding of proteome and identify novel therapeutic targets,\nthis study focused mainly on the differentially abundant proteins\n(DAPs) of <i>Ras</i><sup><i>V12</i></sup>-induced\nGBM. <i>Ras</i><sup><i>V12</i></sup> is a constitutively\nactive Ras mutant form essential for tumor progression by continuously\nactivating signaling pathways leading to uncontrolled tumor growth.\nThis study used a transgenic Drosophila model with <i>Ras</i><sup><i>V12</i></sup> overexpression\nusing the <i>repo-GAL4</i> driver line, specifically in\nglial cells, to study GBM. The high-resolution mass spectrometry (HRMS)-based\nproteomic analysis of the GBM larval central nervous system identified\nthree novel DAPs specific to mitochondria. These DAPs, probable maleylacetoacetate\nisomerase 2 (Q9VHD2), bifunctional methylene tetrahydrofolate dehydrogenase\n(Q04448), and glutamine synthetase1 (P20477), identified through HRMS\nwere further validated by qRT-PCR. The protein–protein interaction\nanalysis revealed interactions between Ras<sup>V12</sup> and DAPs,\nwith functional links to mitochondrial dynamics regulators such as\nDrp1, Marf, Parkin, and HtrA2. Notably, altered expressions of Q9VHD2,\nP20477, and Q04448 were observed during GBM progression, which offers\nnew insights into the involvement of mitochondrial dynamic regulators\nin <i>Ras</i><sup><i>V12</i></sup>-induced GBM\npathophysiology.