Characterization of Lipopolysaccharide Effects on LRRK2 Signaling in RAW Macrophages

Asmaa Oun; Emmy Hoeksema; Ahmed Soliman; Famke Brouwer; Fabiola García-Reyes; Henderikus Pots; Marina Trombetta-Lima; Arjan Kortholt; Amalia M Dolga

doi:10.3390/ijms24021644

Characterization of Lipopolysaccharide Effects on LRRK2 Signaling in RAW Macrophages

Int J Mol Sci. 2023 Jan 13;24(2):1644. doi: 10.3390/ijms24021644.

Authors

Asmaa Oun^{1

2

3}, Emmy Hoeksema¹, Ahmed Soliman^{1

2}, Famke Brouwer¹, Fabiola García-Reyes¹, Henderikus Pots², Marina Trombetta-Lima¹, Arjan Kortholt^{2

4}, Amalia M Dolga¹

Affiliations

¹ Department of Molecular Pharmacology, Faculty of Science and Engineering, Groningen Research Institute of Pharmacy (GRIP), University of Groningen, 9713 AV Groningen, The Netherlands.
² Department of Cell Biochemistry, Groningen Institute of Biomolecular Sciences & Biotechnology (GBB), University of Groningen, 9747 AG Groningen, The Netherlands.
³ Department of Biotechnology, Institute of Graduate Studies and Research, Alexandria University, Alexandria 21526, Egypt.
⁴ YETEM-Innovative Technologies Application and Research Centre, Suleyman Demirel University, 32260 Isparta, Turkey.

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.

Keywords: LPS; LRRK2; cytokines; inflammation.

MeSH terms

Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 / genetics
Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 / metabolism
Lipopolysaccharides* / pharmacology
Macrophages / metabolism
Mutation
Reactive Oxygen Species
Signal Transduction*

Substances

Leucine-Rich Repeat Serine-Threonine Protein Kinase-2
Lipopolysaccharides
Reactive Oxygen Species

Grants and funding

AO is a recipient of Molecular Life and Health Fellowship of the University of Groningen. AMD and AK are recipients of a ParkinsonFonds grant (Grant 1889) and AMD was supported by a Rosalind Franklin Fellowship co-funded by the European Union and the University of Groningen.