Objectives/Goals: This study evaluates whether BT2-mediated reduction of branched-chain amino acids enhances cognitive performance and alters cortical gene expression in transgenic Alzheimer’s disease mouse models, clarifying BCAA’s role in AD pathology and assessing BT2’s potential as a disease-modifying therapy. Methods/Study Population: Four groups of mice were used: wild-type vehicle, wild-type with 20 mg BT2, AD vehicle, and AD with 20 mg BT2(3,6-dichlorobenzobthiophene2-carboxylic acid). BT2, an inhibitor of branched-chain aminotransferase 2, was administered intraperitoneally to reduce BCAA accumulation implicated in mTOR overactivation and tau hyperphosphorylation. Cortical tissue samples were collected for RT-qPCR analysis of 17 genes linked to neurodegeneration. Y-Maze behavioral assays assessed spatial memory and cognition. ANOVA with post-hoc Tukey’s tests (α = 0.05) assessed intergroup significance between behavioral and transcriptional outcomes. Additionally, a Random Forest Regressor Model was utilized in order to identify gene expression patterns predictive of cognitive improvement following BT2 treatment. Results/Anticipated Results: BT2 (3,6-dichlorobenzobthiophene-2-carboxylic acid) treatment restored Beclin-1, VPS-26, NRF2, and NF-κB expression to near wild-type levels, correlating with improved Y-Maze performance. These changes enhanced autophagy, reduced inflammation, and improved mitochondrial resilience. Partial recovery of BACE1, MFN2, and PSD-95 indicated moderate restoration of amyloid processing and synaptic plasticity. Enrichr analysis highlighted pathways in autophagy, mitochondrial biogenesis, and miRNA regulation, suggesting BT2 exerts neuroprotective effects by modulating metabolic and inflammatory homeostasis. The Random Forest Regressor Model’s Feature importance mapping further revealed that changes in Beclin-1, NRF2, and NF-κB expression most strongly correlated with behavioral recovery. Discussion/Significance of Impact: BT2 shows strong potential in mitigating Alzheimer’s pathology through branched-chain amino acid modulation. It normalizes NRF2, Beclin-1, and NF-κB, promoting neuroprotection and cognitive improvement. This supports BT2 as a promising metabolic therapy warranting further long-term studies.
Makam et al. (Wed,) studied this question.