Fig. 8

Validation of proteomic hits reveal both rescue of mutant-driven effects and dysregulatory patterns in long-term MLi-2 treated mice. Follow-up validation of proteomics hits in Western blot confirmed differential changes to endolysosomal, trafficking, and mitochondrial proteins. Within these proteins, we observed divergent patterns of therapeutic and dysregulatory effects in the kidneys of 10-week, MLi-2 treated G2019S KI mice a. Therapeutic trends were observed in non-glycosylated Lamp1, Hgs, and Sfxn3, in which high expression of these proteins in untreated G2019S KI mice were ameliorated down to wildtype levels with LRRK2 inhibition b. We also observed significant increases in glycosylated Lamp1 and Legumain towards levels observed and previously characterized in Lrrk2 KO mice. Additionally, we have shown a significant decrease in Jip4 within the treated mice that were also exacerbated in Lrrk2 KO mice c. Based on these discoveries, we have hypothesized divergent mechanisms for LRRK2 hyperactivity and chronic inhibition d. The proteins in maroon reflect the proteins identified in our proteomics screens (italicized) and validation on WB (not italicized). The grey dashed arrows depict unknown mechanism/relationship with LRRK2 kinase activity. One-way ANOVA with Tukey’s post hoc; n = 6; ****P < 0.0001, **P < 0.005, *P < 0.05; b non-glycosylated Lamp1: F (3, 19)= 13.58, Hgs: F (3, 19)= 13.72, Sfxn3: F (3, 19)= 9.323, c glycosylated Lamp1: F (3, 19)= 13.70, Legumain: F (3, 20)= 61.36, Jip4: F (3, 20)= 27.39