We aimed at gaining more insight into the molecular basis of VWM pathogenesis. Therefore we investigated protein expression patterns in the 2b5ho mouse model using a data-independent mass spectrometry-based quantitative proteomic analysis. The proteome of 4 different brain regions was analyzed at different time points of disease progression. Brain regions were selected based on their regional vulnerability to VWM, and included the cerebellum, corpus callosum, cortex, and brainstem. Commonalities and differences in proteome changes between 2b5ho mouse and VWM patient brains were assessed to determine disease-relevant protein changes during disease development and progression.