Kinase inhibition of G2019S-LRRK2 enhances autolysosome formation and function to reduce endogenous alpha-synuclein intracellular inclusions

Julia Obergasteiger; Giulia Frapporti; Giulia Lamonaca; Sara Pizzi; Anne Picard; Alexandros A Lavdas; Francesca Pischedda; Giovanni Piccoli; Sabine Hilfiker; Evy Lobbestael; Veerle Baekelandt; Andrew A Hicks; Corrado Corti; Peter P Pramstaller; Mattia Volta

doi:10.1038/s41420-020-0279-y

Kinase inhibition of G2019S-LRRK2 enhances autolysosome formation and function to reduce endogenous alpha-synuclein intracellular inclusions

Cell Death Discov. 2020 Jun 8:6:45. doi: 10.1038/s41420-020-0279-y. eCollection 2020.

Authors

Julia Obergasteiger^#¹, Giulia Frapporti^#^{1

2}, Giulia Lamonaca¹, Sara Pizzi¹, Anne Picard¹, Alexandros A Lavdas¹, Francesca Pischedda², Giovanni Piccoli², Sabine Hilfiker³, Evy Lobbestael⁴, Veerle Baekelandt⁴, Andrew A Hicks¹, Corrado Corti¹, Peter P Pramstaller^{1

5

6}, Mattia Volta¹

Affiliations

¹ Institute for Biomedicine, Eurac Research, Affiliated Institute of the University of Lübeck - Via Galvani 31, 39100 Bolzano, Italy.
² Department of Cellular, Computational and Integrative Biology-CIBIO, University of Trento, Via Sommarive 9, 38123 Povo, TN Italy.
³ Department of Anesthesiology, Rutgers University - New Jersey Medical School, Medical Science Building, 185 South Orange Avenue, Newark, NJ 07103 USA.
⁴ Department of Neurosciences, KU Leuven, Herestraat 49 bus 1023, 3000 Leuven, Belgium.
⁵ Department of Neurology, General Central Hospital, Via Böhler 5, 39100 Bolzano, Italy.
⁶ Department of Neurology, University of Lübeck, Ratzeburger Allee 160, 23538 Lübeck, Germany.

^# Contributed equally.

Abstract

The Parkinson's disease (PD)-associated kinase Leucine-Rich Repeat Kinase 2 (LRRK2) is a crucial modulator of the autophagy-lysosome pathway, but unclarity exists on the precise mechanics of its role and the direction of this modulation. In particular, LRRK2 is involved in the degradation of pathological alpha-synuclein, with pathogenic mutations precipitating neuropathology in cellular and animal models of PD, and a significant proportion of LRRK2 patients presenting Lewy neuropathology. Defects in autophagic processing and lysosomal degradation of alpha-synuclein have been postulated to underlie its accumulation and onset of neuropathology. Thus, it is critical to obtain a comprehensive knowledge on LRRK2-associated pathology. Here, we investigated a G2019S-LRRK2 recombinant cell line exhibiting accumulation of endogenous, phosphorylated alpha-synuclein. We found that G2019S-LRRK2 leads to accumulation of LC3 and abnormalities in lysosome morphology and proteolytic activity in a kinase-dependent fashion, but independent from constitutively active Rab10. Notably, LRRK2 inhibition was ineffective upon upstream blockade of autophagosome-lysosome fusion events, highlighting this step as critical for alpha-synuclein clearance.

Keywords: Cellular neuroscience; Macroautophagy.