Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease that primarily affects motor neurons, disrupting muscle function, which eventually leads to death by respiratory failure. TDP-43, a DNA/RNA binding protein that plays numerous roles in RNA processing, has been associated with RNA stress granule pathology in over 95% of ALS cases. TDP-43 has also been linked to dysregulation of specific mRNA targets at both the transcriptional and translational levels. Metabolomic analyses demonstrated that several metabolites in glycolysis and the TCA cycle are altered in Drosophila larvae expressing human TDP-43 compared to controls, suggesting that enzymes within these metabolic pathways are being affected in ALS. To confirm this hypothesis, we are using the GAL4-UAS system in Drosophila to express TDP-43 specifically in the motor neurons or glia. Indeed, transcriptional profiling shows that phosphofructokinase (PFK; Pfk in Drosophila) and glucose-6-phosphate dehydrogenase (G6PD; Zw in Drosophila) expression levels are altered in the context of TDPWT or disease associated TDPG298S. These findings are consistent with increased glycolysis and high levels of pyruvate identified using metabolomics in TDP-43 expressing flies. Current experiments are aimed at identifying additional transcriptional and translational alterations in metabolic pathways that control cellular energetics. Next, we will use genetic approaches to reduce (in the case of upregulation) or increase (in the case of downregulation) the expression levels of metabolic genes in the context of TDP-43. These experiments will determine whether restoring specific targets can rescue TDP-43 dependent phenotypes. This combination of molecular and genetic analyses will establish physiologically significant targets of TDP-43 that can be used for developing novel therapeutic strategies in the future.