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Journal of Neuroinflammation

, 13:295

First Online: 18 November 2016Received: 04 August 2016Accepted: 09 November 2016

Abstract

BackgroundMutations in leucine-rich repeat kinase 2 LRRK2 contribute to both familial and idiopathic forms of Parkinson’s disease PD. Neuroinflammation is a key event in neurodegeneration and aging, and there is mounting evidence of LRRK2 involvement in inflammatory pathways. In a previous study, we described an alteration of the inflammatory response in dermal fibroblasts from PD patients expressing the G2019S and R1441G mutations in LRRK2.

MethodsTaking advantage of cellular reprogramming, we generated induced pluripotent stem cell iPSC lines and neurons thereafter, harboring LRRK2 and LRRK2 mutations. We used gene silencing and functional reporter assays to characterize the effect of the mutations. We examined the temporal profile of TNFα-induced changes in proteins of the NF-κB pathway and optimized western blot analysis to capture α-synuclein dynamics. The effects of the mutations and interventions were analyzed by two-way ANOVA tests with respect to corresponding controls.

ResultsLRRK2 silencing decreased α-synuclein protein levels in mutated neurons and modified NF-κB transcriptional targets, such as PTGS2 COX-2 and TNFAIP3 A20. We next tested whether NF-κB and α-synuclein pathways converged and found that TNFα modulated α-synuclein levels, although we could not detect an effect of LRRK2 mutations, partly because of the individual variability. Nevertheless, we confirmed NF-κB dysregulation in mutated neurons, as shown by a protracted recovery of IκBα and a clear impairment in p65 nuclear translocation in the LRRK2 mutants.

ConclusionsAltogether, our results show that LRRK2 mutations affect α-synuclein regulation and impair NF-κB canonical signaling in iPSC-derived neurons. TNFα modulated α-synuclein proteostasis but was not modified by the LRRK2 mutations in this paradigm. These results strengthen the link between LRRK2 and the innate immunity system underscoring the involvement of inflammatory pathways in the neurodegenerative process in PD.

KeywordsParkinson’s disease LRRK2 Inflammation iPSCs NF-κB α-Synuclein AbbreviationsDADopamine

iPSCInduced pluripotent stem cells

IκBαNuclear factor of kappa light polypeptide gene enhancer in B cells inhibitor, alpha

LRRK2Leucine-rich repeat kinase 2

NF-κBNuclear factor kappa-light-chain-enhancer of activated B cells

PDParkinson’s disease

ROCRas of complex proteins

SNCASynuclein alpha

TNFTumor necrosis factor

TNFRTumor necrosis factor receptor

WTWild-type

Electronic supplementary materialThe online version of this article doi:10.1186-s12974-016-0761-x contains supplementary material, which is available to authorized users.

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Autor: Rakel López de Maturana - Valérie Lang - Amaia Zubiarrain - Amaya Sousa - Nerea Vázquez - Ana Gorostidi - Julio Águila

Fuente: https://link.springer.com/



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