Understanding LRRK2 kinase activity in preclinical models and human subjects through quantitative analysis of LRRK2 and pT73 Rab10
| Autor: | Ceyda Llapashtica, Sarah Huntwork-Rodriguez, Michael T. Maloney, Nathan J. Moerke, Hilda Solanoy, Romeo Maciuca, Anastasia G. Henry, Vitaliy V. Bondar, Shan V. Andrews, Elvira Negrou, Manuel Montalban, Hoang Nguyen, Xiang Wang, Annie Arguello, Laralynne Przybyla |
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| Rok vydání: | 2021 |
| Předmět: |
Science
Parkinson's disease Gene Expression Kinases Disease Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 Peripheral blood mononuclear cell Article Mice medicine Animals Humans Kinase activity Gene Whole blood Enzyme Assays Multidisciplinary Microglia business.industry Kinase LRRK2 nervous system diseases High-Throughput Screening Assays Enzyme Activation medicine.anatomical_structure rab GTP-Binding Proteins Cancer research Leukocytes Mononuclear Medicine business Neuroglia Biomarkers |
| Zdroj: | Scientific Reports Scientific Reports, Vol 11, Iss 1, Pp 1-17 (2021) |
| ISSN: | 2045-2322 |
| Popis: | Variants in the leucine-rich repeat kinase 2 (LRRK2) gene are associated with increased risk for familial and sporadic Parkinson’s disease (PD). Pathogenic variants in LRRK2, including the common variant G2019S, result in increased LRRK2 kinase activity, supporting the therapeutic potential of LRRK2 kinase inhibitors for PD. To better understand the role of LRRK2 in disease and to support the clinical development of LRRK2 inhibitors, quantitative and high-throughput assays to measure LRRK2 levels and activity are needed. We developed and applied such assays to measure the levels of LRRK2 as well as the phosphorylation of LRRK2 itself or one of its substrates, Rab10 (pT73 Rab10). We observed increased LRRK2 activity in various cellular models of disease, including iPSC-derived microglia, as well as in human subjects carrying the disease-linked variant LRRK2 G2019S. Capitalizing on the high-throughput and sensitive nature of these assays, we detected a significant reduction in LRRK2 activity in subjects carrying missense variants in LRRK2 associated with reduced disease risk. Finally, we optimized these assays to enable analysis of LRRK2 activity following inhibition in human peripheral blood mononuclear cells (PBMCs) and whole blood, demonstrating their potential utility as biomarkers to assess changes in LRRK2 expression and activity in the clinic. |
| Databáze: | OpenAIRE |
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