Inhibition of excessive mitochondrial fission reduced aberrant autophagy and neuronal damage caused by LRRK2 G2019S mutation

Hum Mol Genet. 2013 Nov 15;22(22):4545-61. doi: 10.1093/hmg/ddt301. Epub 2013 Jun 27.

Abstract

LRRK2 G2019S mutation is the most common genetic cause of Parkinson's disease (PD). Cellular pathology caused by this mutant is associated with mitochondrial dysfunction and augmented autophagy. However, the underlying mechanism is not known. In this study, we determined whether blocking excessive mitochondrial fission could reduce cellular damage and neurodegeneration induced by the G2019S mutation. In both LRRK2 G2019S-expressing cells and PD patient fibroblasts carrying this specific mutant, treatment with P110, a selective peptide inhibitor of fission dynamin-related protein 1 (Drp1) recently developed in our lab, reduced mitochondrial fragmentation and damage, and corrected excessive autophagy. LRRK2 G2019S directly bound to and phosphorylated Drp1 at Threonine595, whereas P110 treatment abolished this phosphorylation. A site-directed mutant, Drp1(T595A), corrected mitochondrial fragmentation, improved mitochondrial mass and suppressed excessive autophagy in both cells expressing LRRK2 G2019S and PD patient fibroblasts carrying the mutant. Further, in dopaminergic neurons derived from LRRK2 G2019S PD patient-induced pluripotent stem cells, we demonstrated that either P110 treatment or expression of Drp1(T595A) reduced mitochondrial impairment, lysosomal hyperactivity and neurite shortening. Together, we propose that inhibition of Drp1-mediated excessive mitochondrial fission might be a strategy for treatment of PD relevant to LRRK2 G2019S mutation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autophagy / drug effects*
  • Autophagy / genetics
  • Dynamins
  • Enzyme Inhibitors / pharmacology*
  • Female
  • GTP Phosphohydrolases / antagonists & inhibitors
  • GTP Phosphohydrolases / pharmacology*
  • Guanine / metabolism
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Leucine-Rich Repeat Serine-Threonine Protein Kinase-2
  • Male
  • Microtubule-Associated Proteins / antagonists & inhibitors
  • Middle Aged
  • Mitochondria / metabolism
  • Mitochondria / pathology
  • Mitochondrial Dynamics / drug effects*
  • Mitochondrial Dynamics / genetics
  • Mitochondrial Proteins / antagonists & inhibitors
  • Mitochondrial Proteins / genetics
  • Molecular Targeted Therapy
  • Mutagenesis, Site-Directed
  • Mutation
  • Neurites / pathology
  • Neuroprotective Agents / pharmacology*
  • Parkinson Disease / genetics
  • Parkinson Disease / physiopathology*
  • Peptide Fragments / pharmacology*
  • Phosphorylation
  • Protein Serine-Threonine Kinases / genetics*
  • Serine / metabolism

Substances

  • Enzyme Inhibitors
  • Microtubule-Associated Proteins
  • Mitochondrial Proteins
  • Neuroprotective Agents
  • P110 peptide
  • Peptide Fragments
  • Serine
  • Guanine
  • LRRK2 protein, human
  • Leucine-Rich Repeat Serine-Threonine Protein Kinase-2
  • Protein Serine-Threonine Kinases
  • GTP Phosphohydrolases
  • DNM1L protein, human
  • Dynamins