The striatal kinase DCLK3 produces neuroprotection against mutant huntingtin

Galvan, Laurie; Francelle, Laetitia; Gaillard, Marie-Claude; Longprez, Lucie de; Sauvage, María-Angeles Carrillo-de; Liot, Géraldine; Cambon, Karine; Stimmer, Lev; Luccantoni, Sophie; Flament, Julien; Valette, Julien; Chaldée, Michel de; Aurégan, Gwenaëlle; Guillermier, Martine; Joséphine, Charlène; Petit, Fanny; Jan, Caroline; Jarrige, Margot; Dufour, Noëlle; Bonvento, Gilles; Humbert, Sandrine; Saudou, Frédéric; Hantraye, Philippe; Mérienne, Karine; Bemelmans, Alexis-Pierre; Perrier, Anselme L.; Déglon, Nicole; Brouillet, Emmanuel

doi:10.1093/brain/awy057

Cited by 29 publications

(25 citation statements)

References 57 publications

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“…The reduction in tNAA levels in mouse models is coincident with decreased levels of mRNA and proteins that are considered molecular markers of the striatum, ie preferentially expressed in striatal medium sized spiny neurons 33,34 . For example, reduced levels of the mRNA and/or protein of striatal markers such as dopamine, 3′,5′‐cyclic adenosine monophosphate‐regulated neuronal phosphoprotein, phosphodiesterase 10 or the recently studied doublecortin‐like kinase 3 have been observed in Ki140CAG and R6/1 mice 33–36 . The levels of these markers have also been found to be reduced in post‐mortem samples in HD patients 36 .…”

Section: Discussionmentioning

confidence: 99%

“…For example, reduced levels of the mRNA and/or protein of striatal markers such as dopamine, 3′,5′‐cyclic adenosine monophosphate‐regulated neuronal phosphoprotein, phosphodiesterase 10 or the recently studied doublecortin‐like kinase 3 have been observed in Ki140CAG and R6/1 mice 33–36 . The levels of these markers have also been found to be reduced in post‐mortem samples in HD patients 36 . In spite of the absence of neuronal loss at 12 months, both models showed behavioral phenotypes 19,22,31,32 .…”

Section: Discussionmentioning

confidence: 99%

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Complementarity of gluCEST and ¹H‐MRS for the study of mouse models of Huntington's disease

Pépin

Longprez

Trovero³

et al. 2020

NMR in Biomedicine

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Identification of relevant biomarkers is fundamental to understand biological processes of neurodegenerative diseases and to evaluate therapeutic efficacy. Atrophy of brain structures has been proposed as a biomarker, but it provides little information about biochemical events related to the disease. Here, we propose to identify early and relevant biomarkers by combining biological specificity provided by 1 H-MRS and high spatial resolution offered by gluCEST imaging. For this, two different genetic mouse models of Huntington's disease (HD)-the Ki140CAG model, characterized by a slow progression of the disease, and the R6/1 model, which mimics the juvenile form of HD-were used. Animals were scanned at 11.7 T using a protocol combining 1 H-MRS and gluCEST imaging. We measured a significant decrease in levels of N-acetyl-aspartate, a metabolite mainly located in the neuronal compartment, in HD animals, and the decrease seemed to be correlated with disease severity.In addition, variations of tNAA levels were correlated with striatal volumes in both models. Significant variations of glutamate levels were also observed in Ki140CAG but not in R6/1 mice. Thanks to its high resolution, gluCEST provided complementary insights, and we highlighted alterations in small brain regions such as the corpus callosum in Ki140CAG mice, whereas the glutamate level was unchanged in the whole brain of R6/1 mice. In this study, we showed that 1 H-MRS can provide key information about biological processes occurring in vivo but was limited by the spatial resolution. On the other hand, gluCEST may finely point to alterations in unexpected brain regions, but it can also be blind to disease processes when glutamate levels are preserved. This highlights in a practical context the complementarity of the two methods to study animal models of neurodegenerative diseases and to identify relevant biomarkers.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Complementarity of gluCEST and ¹H‐MRS for the study of mouse models of Huntington's disease

Pépin

Longprez

Trovero³

et al. 2020

NMR in Biomedicine

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“…Fractionation experiments. For fractionation experiments, samples were processed as previously described 32 . Briefly, frozen pieces of tissue were weighted and placed in a loose-fitting Dounce homogenizer with 25 volumes of ice-cold Buffer A [10 mM HEPES pH 7.9, 10 mM KCl, 0.1 mM EDTA, 0.1 mM EGTA, 1 mM DTT, 1X Halt protease and phosphatase inhibitor cocktail (Thermo Scientific)] and homogenized with 10 up-and-down passes.…”

Section: Methodsmentioning

confidence: 99%

Inhibition of HDAC6 activity protects dopaminergic neurons from alpha-synuclein toxicity

Francelle

Outeiro

Rappold

2020

Sci Rep

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The neuropathological hallmarks of Parkinson's disease include preferential vulnerability of dopaminergic neurons of the substantia nigra pars compacta, and accumulation of intraneuronal protein inclusions known as Lewy bodies. These inclusions contain, among other proteins, aggregated alpha-synuclein and histone deacetylase 6 (HDAC6). In our study we found that selective inhibition of HDAC6 activity by Tubastatin A has protective effects in a rat model of Parkinson's disease. We provide evidence that this protection may be due to the activation of chaperone-mediated autophagy through the up-regulation of key members of this pathway. Moreover, Tubastatin A significantly inhibited the expression of a toxic form of alpha-synuclein that is phosphorylated at serine position 129. Tubastatin A treatment also permitted to partially modulate neuroinflammation. Taken together, our study highlights the neuroprotective effects of Tubastatin A in a rat model of Parkinson's disease and provides mechanistic insight in Tubastatin A-mediated protection against alpha-synuclein toxicity and substantia nigra degeneration. These findings are of potential therapeutic value in Parkinson's disease and other synucleinopathies.www.nature.com/scientificreports www.nature.com/scientificreports/ induced by human alpha-synuclein overexpression in the substantia nigra pars compacta 15 . We already used this rat model to test the effect of other putative therapeutic targets, such as sirtuin 2 (SIRT2) 8 . We adapted the experimental design in order to phenocopy early-onset PD defects. To specifically inhibit HDAC6, we used Tubastatin A and tested it as a preventive drug in this early onset model of PD. We found that inhibition of HDAC6 activity by Tubastatin A has a disease modifier effect in the substantia nigra pars compacta by protecting dopaminergic neurons against alpha-synuclein toxicity, accompanied by reduced astrogliosis. Our results demonstrate that Tubastatin A treatment lead to differential protein expression compared to the vehicle. Interestingly, we found that Hsc70 and Lamp2 proteins were upregulated by Tubastatin A, whereas alpha-synuclein was downregulated. Importantly, we did not detect phosphorylation of alpha-synuclein at serine position 129, an accepted marker of pathological alpha-synuclein, in the rats treated with Tubastatin A. Overall, our work supports the hypothesis that specific inhibition of HDAC6 activity increases alpha-synuclein acetylation, up-regulates key members of the chaperone-mediated autophagy, and reduces alpha-synuclein expression and toxicity. Therefore, HDAC6 is a potential therapeutic target for PD and other alpha-synucleinopathies. Scientific RepoRtS |(2020) 10:6064 | https://doi.CM3050S) at a 30 µm thickness. Sections were collected as serial coronal free-floating sections in a storage solution (0.1 M PBS; 0.1% sodium azide), stored at 4 °C until use.Scientific RepoRtS | (2020) 10:6064 | https://doi.

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“…Using a different approach with lentiviral vectors in mice, we tested whether the different forms of ΔLRRK2 could modify the neurotoxicity produced by the Nterminal domain of human huntingtin (Htt) with a pathological expansion of its poly-glutamine (Q) region (Htt-N171-82Q) [30]. Lentiviral vectors were injected into the striatum of wildtype mice to produce local cell loss within the six weeks following transduction, as previously described [31,32].…”

Section: Effects Of Co-transduction With Aav-α-syn A53t and Aav-∆lrrkmentioning

confidence: 99%

The C-terminal fragment of LRRK2 with the G2019S substitution increases the neurotoxicity of mutant A53T α-synuclein in dopaminergic neurons in vivo

Cresto

Gardier

Gaillard

et al. 2020

Preprint

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Background: Alpha-synuclein (α-syn) and leucine-rich repeat kinase 2 (LRRK2) likely play crucial roles both in sporadic and familial forms of Parkinson’s disease (PD). The most prevalent mutation in LRRK2 is the G2019S substitution, which induces neurotoxicity through increased kinase activity. There is likely an interplay between LRRK2 and α-syn involved in the neurodegeneration of dopaminergic (DA) neurons in the substantia nigra (SNc) in PD. However, the mechanisms underlying this interplay are ill-defined. Here, we investigated whether LRRK2G2019S can increase the neurotoxicity induced by a mutant form of α-syn (A53T mutation) in DA neuronsin vivo.Methods: We used a co-transduction approach with AAV2/6 vectors encoding human a-synA53T and the C-terminal portion of LRRK2 (ΔLRRK2), which contains the kinase domain, with either the G2019S mutation (ΔLRRK2G2019) alone or the D1994A mutation (ΔLRRK2G2019S/D1994A), which inactivates the kinase activity of LRRK2. The AAVs were co-injected into the rat SNc and histological evaluation was performed at 6- and 15-weeks post-injection (PI). Results: Most SNc neurons co-expressed ΔLRRK2 and human α-synA53T after transduction. ΔLRRK2G2019S alone produced no cell loss at 15-weeks PI, whereas as expected, transduction with AAV-a-synA53T mixed with a control AAV coding for GFP produced significant loss of DA neurons. Co-injection of AAV-ΔLRRK2G2019S and AAV-α-synA53T induced a loss of DA neurons slightly but significantly greater than that produced by co-injection of AAV-α-synA53T and AAV-GFP. We also studied the inactive form, ΔLRRK2G2019S/D1994A at 6 weeks PI. Results showed that ΔLRRK2G2019S/D1994A did not alter the early toxicity of α-synA53T, in contrast to the active form ΔLRRK2G2019S that produced a moderate but significant increase in α-synA53T toxicity.Conclusion. Thus, these results show that mutant LRRK2 may selectively facilitate α-syn toxicity in DA neurons through a cell-autonomous mechanism involving its kinase activity. However, considering that the effect of ΔLRRK2G2019S upon human α-synA53T is moderate in our paradigm where pathological proteins are overexpressed, the study supports the hypothesis that the interplay between LRRK2 and α-syn also implicates non-cell-autonomous mechanisms such as those involved in neuroinflammation.

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The striatal kinase DCLK3 produces neuroprotection against mutant huntingtin

Cited by 29 publications

References 57 publications

Complementarity of gluCEST and ¹H‐MRS for the study of mouse models of Huntington's disease

Complementarity of gluCEST and ¹H‐MRS for the study of mouse models of Huntington's disease

Inhibition of HDAC6 activity protects dopaminergic neurons from alpha-synuclein toxicity

The C-terminal fragment of LRRK2 with the G2019S substitution increases the neurotoxicity of mutant A53T α-synuclein in dopaminergic neurons in vivo

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The striatal kinase DCLK3 produces neuroprotection against mutant huntingtin

Cited by 29 publications

References 57 publications

Complementarity of gluCEST and 1H‐MRS for the study of mouse models of Huntington's disease

Complementarity of gluCEST and 1H‐MRS for the study of mouse models of Huntington's disease

Inhibition of HDAC6 activity protects dopaminergic neurons from alpha-synuclein toxicity

The C-terminal fragment of LRRK2 with the G2019S substitution increases the neurotoxicity of mutant A53T α-synuclein in dopaminergic neurons in vivo

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Complementarity of gluCEST and ¹H‐MRS for the study of mouse models of Huntington's disease

Complementarity of gluCEST and ¹H‐MRS for the study of mouse models of Huntington's disease