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Published on: December 14, 2017
Characterization of Lipopolysaccharide Effects on LRRK2 Signaling in RAW Macrophages
Asmaa Oun1,2,3, Emmy Hoeksema1, Ahmed Soliman1,2
1Department of Molecular Pharmacology, Faculty of Science and Engineering, Groningen Research Institute of Pharmacy (GRIP), University of Groningen, 9713 AV Groningen, The Netherlands.
Abstract:
Dysfunction of the immune system and mitochondrial metabolism has been associated with Parkinson's disease (PD) pathology. Mutations and increased kinase activity of leucine-rich repeat kinase 2 (LRRK2) are linked to both idiopathic and familial PD. However, the function of LRRK2 in the immune cells under inflammatory conditions is contradictory. Our results showed that lipopolysaccharide (LPS) stimulation increased the kinase activity of LRRK2 in parental RAW 264.7 (WT) cells. In addition to this, LRRK2 deletion in LRRK2 KO RAW 264.7 (KO) cells altered cell morphology following LPS stimulation compared to the WT cells, as shown by an increase in the cell impedance as observed by the xCELLigence measurements. LPS stimulation caused an increase in the cellular reactive oxygen species (ROS) levels in both WT and KO cells. However, WT cells displayed a higher ROS level compared to the KO cells. Moreover, LRRK2 deletion led to a reduction in interleukin-6 (IL-6) inflammatory cytokine and cyclooxygenase-2 (COX-2) expression and an increase in lactate production after LPS stimulation compared to the WT cells. These data illustrate that LRRK2 has an effect on inflammatory processes in RAW macrophages upon LPS stimulation.
Insights
Leucine-rich repeat kinase 2 (LRRK2) influences immune cell responses in Parkinson's disease models. Its deletion in macrophages reduced inflammation and altered metabolism following lipopolysaccharide stimulation.
Area of Science:
- Immunology
- Neuroscience
- Cell Biology
Background:
- Immune system and mitochondrial metabolism dysfunction are implicated in Parkinson's disease (PD).
- Mutations and altered kinase activity of leucine-rich repeat kinase 2 (LRRK2) are linked to PD.
- The role of LRRK2 in immune cells during inflammation remains unclear.
Purpose of the Study:
- To investigate the function of LRRK2 in RAW 264.7 macrophages under inflammatory conditions.
- To determine the impact of LRRK2 deletion on cellular responses to lipopolysaccharide (LPS).
Main Methods:
- Utilized wild-type (WT) and LRRK2 knockout (KO) RAW 264.7 macrophage cell lines.
- Stimulated cells with lipopolysaccharide (LPS) to induce inflammation.
- Measured cell morphology (cell impedance via xCELLigence), reactive oxygen species (ROS) levels, interleukin-6 (IL-6) and cyclooxygenase-2 (COX-2) expression, and lactate production.
Main Results:
- LPS stimulation increased LRRK2 kinase activity in WT cells.
- LRRK2 deletion altered cell morphology and increased cell impedance post-LPS stimulation.
- While both WT and KO cells showed increased ROS, WT cells had higher levels.
- LRRK2 deletion reduced IL-6 and COX-2 expression but increased lactate production after LPS stimulation.
Conclusions:
- LRRK2 plays a significant role in regulating inflammatory processes and metabolic changes in macrophages.
- These findings suggest LRRK2 as a potential therapeutic target for modulating neuroinflammation in Parkinson's disease.
