Targeting Rab‐RILPL interactions as a strategy to downregulate pathogenic LRRK2 in Parkinson's disease

Familial Parkinson's disease (PD) is frequently linked to multiple disease‐causing mutations within Leucine‐Rich Repeat Protein Kinase 2 (LRRK2), leading to aberrant kinase activity. Multiple pathogenic effects of enhanced LRRK2 activity have been identified, including loss of cilia and centrosomal cohesion defects. When phosphorylated by LRRK2, Rab8a and Rab10 bind to phospho‐specific RILPL effector proteins. RILPL‐mediated accumulation of pRabs proximal to the mother centriole is critical for initiating deficits in ciliogenesis and centrosome cohesion mediated by LRRK2. We hypothesized that Rab‐derived phospho‐mimics may serve to block phosphorylated Rab proteins from docking with RILPL in the context of hyperactive LRRK2 mutants. This would serve as an alternative strategy to downregulate pathogenic signaling mediated by LRRK2, rather than targeting LRRK2 kinase activity itself. To test this theory, we designed a series of constrained peptides mimicking phosphorylated Switch II derived from Rab8. These RILPL interacting peptides, termed RIP, were further shown to permeate cells. Further, several peptides were found to bind RILPL2 and restore ciliogenesis and centrosomal cohesion defects in cells expressing PD‐associated mutant LRRK2. This research demonstrates the utility of constrained peptides as downstream inhibitors to target pathogenic LRRK2 activity and may provide an alternative approach to target specific pathways activated by LRRK2. A series of constrained peptides mimicking phosphorylated Switch II derived from Rab8 were designed. These RILPL‐interacting peptides, termed RIP, were shown to permeate cells, and several peptides were found to bind RILPL2 and restore ciliogenesis and centrosomal cohesion defects in cells expressing Parkinson's disease‐associated mutant LRRK2 (Leucine‐Rich Repeat Protein Kinase 2).