Neuroprotective Effect of the LRRK2 Kinase Inhibitor PF-06447475 in Human Nerve-Like Differentiated Cells Exposed to Oxidative Stress Stimuli: Implications for Parkinson’s Disease

Abstract

Leucine-rich repeat kinase 2 (LRRK2) has been implicated in oxidative stress (OS) and neurodegeneration in Parkinson's disease (PD). However, the pathophysiological mechanism of the LRRK2 kinase in neurons under stress stimuli is not yet understood. We demonstrate that rotenone (ROT), a mitochondria complex I inhibitor frequently used to generate in vitro and in vivo experimental models of PD, induces LRRK2 phosphorylation at serine 935 p-(S935) concomitant with cell death in nerve-like differentiated cells (NLCs). Indeed, ROT (50 µM) at 6 h exposure significantly increased reactive oxygen species (ROS) (~100 %), p-(S935)-LRRK2 kinase [~2 f(old)-(i)ncrease] level, induced nuclei condensation/fragmentation (16 %), increased the expression of NF-κB (5.6 f-i), p53 (5.3 f-i), c-Jun (5.4 f-i) transcription factors, activated caspase-3 (8.0 f-i) and AIF (6.8 f-i) proteins; but significantly decreased mitochondrial membrane potential (∆Ψ, ~21 %), indicative of apoptosis -a type of regulated cell death process- compared to untreated cells. Strikingly, the LRRK2 kinase inhibitor PF-06447475 (PF-475, 1 µM) protects NLCs against ROT induced noxious effect. The inhibitor not only blocked the p-(S935)-LRRK2 kinase phosphorylation but also completely abolished ROS, and significantly reversed all ROT-induced apoptosis signaling and OS associated markers to comparable control values. We conclude that wild-type LRRK2 may act as a pro-apoptotic factor under OS stimuli. Our findings suggest an association between OS and LRRK2 phosphorylation in the NLCs death process, as PD model. Therefore, the pharmacological inhibition of LRRK2 might help to understand the OS-mediated kinase activation in PD neurodegenerative disorder.

GSK-3β Inhibitor Alsterpaullone Attenuates MPP+-Induced Cell Damage in a c-Myc-Dependent Manner in SH-SY5Y Cells

“…Mitochondria are important intercellular organelles in the nerve cells, producing energy through the mitochondrial respiratory chain and acting as a mediator of the cell survival and death (Keilhoff et al, 2016 ; Mendivil-Perez et al, 2016 ). Accumulating studies of PD animals and PD patients suggested that mitochondrial dysfunction may play crucial roles in the PD pathogenesis (Martin, 2006 ; Desmet et al, 2017 ).…”

Leucine-Rich Repeat Kinase 2 in Parkinson’s Disease: Updated from Pathogenesis to Potential Therapeutic Target

“…Rotenone (ROT) introduction into nerve-like differentiated cells generates in-vitro models of PD, resulting in LRRK2 phosphorylation at Ser935, nuclear condensation, increased nuclear factor κ-light-chain-enhancer of activated B (NF-κB) expression, and activated caspase-3. Subsequent exposure to PF-06447475 blocks LRRK2 phosphorylation at Ser935 and reverses the apoptotic signaling activated by ROT [66]. In 2 preclinical mouse models of LRRK2 (bacterial artificial chromosome transgenic hG2019S and hR1441G), GNE-7915 was reported to enhance DA release and synaptic vesicle recycling [67].…”

Cholinergic-like neurons carrying PSEN1 E280A mutation from familial Alzheimer’s disease reveal intraneuronal Aβ42 peptide accumulation, hyperphosphorylation of TAU, oxidative stress, apoptosis and Ca2+ flux dysregulation: Therapeutic Implications

“…We first evaluated whether the PSEN1 E280A mutation affects the capacity of MSCs to generate mesodermal and ectodermal lineages through differentiation and transdifferentiation, respectively. Therefore, WJ-MSCs were isolated from ten umbilical cords of volunteers according to standard procedures [39], and PCR-RFLP electrophoretic profile analysis identified one umbilical cord sample out of ten as a carrier of the mutation PSEN1 c.839A > C, p.E280A (PSEN1 E280A MSCs). Then, PSEN1 E280A MSCs and wild-type WJ-MSCs (WT PSEN1 MSCs) were further cultured and characterized for morphological, karyotype, immuno-phenotypic features and differentiation capabilities.…”