Measuring LRRK2 Phosphorylation by Flow Cytometry
LRRK2 Pharmacodynamic Assays, 2013
Objective/Rationale: † † † † † ††
There is a possibility that inhibitors of the leucine-rich repeat kinase 2 (LRRK2) protein could be used for treating Parkinsonís disease. Our projects aims to develop a test that uses blood samples to measure how well LRRK2 inhibitors are working in human cells.
Project Description: † † † † † † †
Antibodies that can be used to measure LRRK2 will be modified and optimized for use in a process called flow cytometry. Blood samples will then be collected from control and Parkinsonís disease patients and the blood will be treated with LRRK2 inhibitors. Flow cytometry will be used to measure the phosphorylation (process that turns protein enzymes on and off) of LRRK2 in different types of white blood cells. We aim to further validate our findings that inhibitors of LRRK2 will result in less phosphorylation being detected.
Relevance to Diagnosis/Treatment of Parkinsonís Disease: † † † † † † † † † †
There is a possibility that inhibitors of the leucine-rich repeat kinase 2 (LRRK2) protein could be used for treating Parkinsonís disease however, little is known about what LRRK2 actually does. Therefore, it is very difficult to know what to measure in order to demonstrate that inhibitors of LRRK2 are working. If successful, our assay will provide a reliable readout to determine if LRRK2 kinase inhibitors are indeed, effectively inhibiting LRRK2 in human cells.†
Anticipated Outcome: † † † † ††
Human blood consists of a number of different cell types. Many of these express the LRRK2 protein. We anticipate generating a robust and reliable assay to measure LRRK2 simultaneously in a number of different blood cell types. This will tell us more about which cells express LRRK2 and if there are any differences seen in Parkinsonís disease. The assay could also demonstrate that a blood test could be used to determine that LRRK2 inhibitors are working.
Senior Research Fellow at The University of Sydney
Location: Sydney, NSW, Australia