[HTML][HTML] Impaired inflammatory responses in murine Lrrk2-knockdown brain microglia

B Kim, MS Yang, D Choi, JH Kim, HS Kim, W Seol… - PloS one, 2012 - journals.plos.org
B Kim, MS Yang, D Choi, JH Kim, HS Kim, W Seol, S Choi, I Jou, EY Kim, E Joe
PloS one, 2012journals.plos.org
LRRK2, a Parkinson's disease associated gene, is highly expressed in microglia in addition
to neurons; however, its function in microglia has not been evaluated. Using Lrrk2
knockdown (Lrrk2-KD) murine microglia prepared by lentiviral-mediated transfer of Lrrk2-
specific small inhibitory hairpin RNA (shRNA), we found that Lrrk2 deficiency attenuated
lipopolysaccharide (LPS)-induced mRNA and/or protein expression of inducible nitric oxide
synthase, TNF-α, IL-1β and IL-6. LPS-induced phosphorylation of p38 mitogen-activated …
LRRK2, a Parkinson's disease associated gene, is highly expressed in microglia in addition to neurons; however, its function in microglia has not been evaluated. Using Lrrk2 knockdown (Lrrk2-KD) murine microglia prepared by lentiviral-mediated transfer of Lrrk2-specific small inhibitory hairpin RNA (shRNA), we found that Lrrk2 deficiency attenuated lipopolysaccharide (LPS)-induced mRNA and/or protein expression of inducible nitric oxide synthase, TNF-α, IL-1β and IL-6. LPS-induced phosphorylation of p38 mitogen-activated protein kinase and stimulation of NF-κB-responsive luciferase reporter activity was also decreased in Lrrk2-KD cells. Interestingly, the decrease in NF-κB transcriptional activity measured by luciferase assays appeared to reflect increased binding of the inhibitory NF-κB homodimer, p50/p50, to DNA. In LPS-responsive HEK293T cells, overexpression of the human LRRK2 pathologic, kinase-active mutant G2019S increased basal and LPS-induced levels of phosphorylated p38 and JNK, whereas wild-type and other pathologic (R1441C and G2385R) or artificial kinase-dead (D1994A) LRRK2 mutants either enhanced or did not change basal and LPS-induced p38 and JNK phosphorylation levels. However, wild-type LRRK2 and all LRRK2 mutant variants equally enhanced NF-κB transcriptional activity. Taken together, these results suggest that LRRK2 is a positive regulator of inflammation in murine microglia, and LRRK2 mutations may alter the microenvironment of the brain to favor neuroinflammation.
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