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Sci Rep. 2017 Aug 31;7(1):10300. doi: 10.1038/s41598-017-10501-z.

Selective LRRK2 kinase inhibition reduces phosphorylation of endogenous Rab10 and Rab12 in human peripheral mononuclear blood cells.

Author information

1
Neurodegeneration and Biologics, H. Lundbeck A/S, Ottiliavej 9, Valby, DK-2500, Denmark.
2
Evotec (München) GmbH, Am Klopferspitz 19a, 82152, Martinsried, Germany.
3
Vernalis (R&D) Ltd, Granta Park, Great Abington, Cambridge, CB21 6GB, United Kingdom.
4
Discovery Chemistry, H. Lundbeck A/S, Ottiliavej 9, DK-2500, Valby, Denmark.
5
Neurodegeneration and Biologics, H. Lundbeck A/S, Ottiliavej 9, Valby, DK-2500, Denmark. kvc@lundbeck.com.

Abstract

Genetic variation in the leucine-rich repeat kinase 2 (LRRK2) gene is associated with risk of familial and sporadic Parkinson's disease (PD). To support clinical development of LRRK2 inhibitors as disease-modifying treatment in PD biomarkers for kinase activity, target engagement and kinase inhibition are prerequisite tools. In a combined proteomics and phosphoproteomics study on human peripheral mononuclear blood cells (PBMCs) treated with the LRRK2 inhibitor Lu AF58786 a number of putative biomarkers were identified. Among the phospho-site hits were known LRRK2 sites as well as two phospho-sites on human Rab10 and Rab12. LRRK2 dependent phosphorylation of human Rab10 and human Rab12 at positions Thr73 and Ser106, respectively, was confirmed in HEK293 and, more importantly, Rab10-pThr73 inhibition was validated in immune stimulated human PBMCs using two distinct LRRK2 inhibitors. In addition, in non-stimulated human PBMCs acute inhibition of LRRK2 with two distinct LRRK2 inhibitor compounds reduced Rab10-Thr73 phosphorylation in a concentration-dependent manner with apparent IC50's equivalent to IC50's on LRRK2-pSer935. The identification of Rab10 phosphorylated at Thr73 as a LRRK2 inhibition marker in human PBMCs strongly support inclusion of assays quantifying Rab10-pThr73 levels in upcoming clinical trials evaluating LRRK2 kinase inhibition as a disease-modifying treatment principle in PD.

PMID:
28860483
PMCID:
PMC5578959
DOI:
10.1038/s41598-017-10501-z
[Indexed for MEDLINE]
Free PMC Article

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