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Predictive Markers of LRRK2 Inhibition in Biofluids

22 Pages Posted: 12 Oct 2019

See all articles by Shijie Wang

Shijie Wang

Duke University - Center for Neurodegeneration and Neurotherapeutics

Kaela Kelly

Duke University - Center for Neurodegeneration and Neurotherapeutics

Nathalie Schussler

Sanofi - France

Sylviane Boularand

Sanofi - France

Laurent Dubois

Sanofi - France

Frank Hsieh

Nextcea, Inc.

Elizabeth Tengstrand

Nextcea, Inc.

Jonathan M. Brotchie

Atuka Inc.

James B. Koprich

Atuka Inc.

Andrew B. West

Duke University - Center for Neurodegeneration and Neurotherapeutics

More...

Abstract

Background: Hyper-activated LRRK2 is linked to Parkinson’s disease susceptibility and progression. Quantitative measures of LRRK2 inhibition, especially in the brain, may be critical in the development of successful LRRK2- targeting therapeutics.

Methods: Three structurally distinct, brain-penetrant, and selective LRRK2 small-molecule kinase inhibitors (PFE360, MLi2, and RA283) were orally administered to groups of cynomolgus macaques at different doses. Biofluid markers with proposed predictive value for assessing LRRK2 inhibition were measured from samples of blood, urine, and cerebral-spinal fluid (CSF).

Findings: LRRK2 kinase inhibition led to consistent reduced pS935-LRRK2 and pRab10 proteins normalized to β- actin in blood mononuclear cells, reduced exosome-total LRRK2 protein normalized to TSG101 and didocosahexaenoyl (22:6) bis (monoacylglycerol) phosphate normalized to creatinine in urine, and reduced exosometotal LRRK2 and autophosphorylated pS1292-LRRK2 protein normalized to flotillin in CSF. Incomplete LRRK2 kinase inhibition reduced LRRK2 protein secretion in exosomes, whereas high drug exposures reduced both exosome and cellular levels of LRRK2 protein.

Interpretation: Orthogonal pairs of markers for LRRK2 inhibition were identified in urine and CSF that can be used in combination with blood markers to non-invasively monitor the potency of LRRK2-targeting therapeutics in the brain and periphery.

Funding Statement: Research support was provided by NIH U01 NS097028, R01 NS064934, and experiments were further supported through internal research programs at Atuka Inc, Sanofi, and Nextcea Inc.

Declaration of Interests: S.W., K.K., and A.B.W. declare no competing conflicts of interest. L.D., S.B., and N.S. are employees of Sanofi and have shares in Sanofi. J.B.K. and J.M.B. are employees of Atuka and have shares in Atuka Inc. F.H. and E.T. are employed by Nexctea which holds patent rights to the di-22:6-BMP biomarker for neurological diseases involving lysosomal dysfunction (US 8,313,949, Japan 5,702,363, and Europe EP2419742).

Ethics Approval Statement: All procedures were performed in full compliance with standards for the care and use of laboratory animals, according to French and European Community (Directive 2010/63/EU) legislation, and approved by the local Animal Ethics Committee and the French Ministry for Research, under authorization H94-002-4 arrêté 2015-160.

Keywords: Parkinson's disease, leucine-rich repeat kinase 2 (LRRK2), Park8, small-molecule kinase inhibitors, Pharmacodynamic biomarkers

Suggested Citation

Wang, Shijie and Kelly, Kaela and Schussler, Nathalie and Boularand, Sylviane and Dubois, Laurent and Hsieh, Frank and Tengstrand, Elizabeth and Brotchie, Jonathan M. and Koprich, James B. and West, Andrew B., Predictive Markers of LRRK2 Inhibition in Biofluids (October 9, 2019). Available at SSRN: https://ssrn.com/abstract=3466981 or http://dx.doi.org/10.2139/ssrn.3466981

Shijie Wang

Duke University - Center for Neurodegeneration and Neurotherapeutics

100 Fuqua Drive
Durham, NC 27715
United States

Kaela Kelly

Duke University - Center for Neurodegeneration and Neurotherapeutics

100 Fuqua Drive
Durham, NC 27715
United States

Nathalie Schussler

Sanofi - France

France

Sylviane Boularand

Sanofi - France

France

Laurent Dubois

Sanofi - France

France

Frank Hsieh

Nextcea, Inc.

Woburn, MA
United States

Elizabeth Tengstrand

Nextcea, Inc.

Woburn, MA
United States

Jonathan M. Brotchie

Atuka Inc.

Toronto
Canada

James B. Koprich

Atuka Inc.

Toronto
Canada

Andrew B. West (Contact Author)

Duke University - Center for Neurodegeneration and Neurotherapeutics ( email )

100 Fuqua Drive
Durham, NC 27715
United States

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