Kv1.3 modulates neuroinflammation and neurodegeneration in Parkinson’s disease
Souvarish Sarkar, Hai M. Nguyen, Emir Malovic, Jie Luo, Monica Langley, Bharathi N. Palanisamy, Neeraj Singh, Sireesha Manne, Matthew Neal, Michelle Gabrielle, Ahmed Abdalla, Poojya Anantharam, Dharmin Rokad, Nikhil Panicker, Vikrant Singh, Muhammet Ay, Adhithiya Charli, Dilshan Harischandra, Lee-Way Jin, Huajun Jin, Srikant Rangaraju, Vellareddy Anantharam, Heike Wulff, Anumantha G. Kanthasamy
Souvarish Sarkar, Hai M. Nguyen, Emir Malovic, Jie Luo, Monica Langley, Bharathi N. Palanisamy, Neeraj Singh, Sireesha Manne, Matthew Neal, Michelle Gabrielle, Ahmed Abdalla, Poojya Anantharam, Dharmin Rokad, Nikhil Panicker, Vikrant Singh, Muhammet Ay, Adhithiya Charli, Dilshan Harischandra, Lee-Way Jin, Huajun Jin, Srikant Rangaraju, Vellareddy Anantharam, Heike Wulff, Anumantha G. Kanthasamy
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Research Article Inflammation Neuroscience

Kv1.3 modulates neuroinflammation and neurodegeneration in Parkinson’s disease

Abstract

Characterization of the key cellular targets contributing to sustained microglial activation in neurodegenerative diseases, including Parkinson’s disease (PD), and optimal modulation of these targets can provide potential treatments to halt disease progression. Here, we demonstrated that microglial Kv1.3, a voltage-gated potassium channel, was transcriptionally upregulated in response to aggregated α-synuclein (αSynAgg) stimulation in primary microglial cultures and animal models of PD, as well as in postmortem human PD brains. Patch-clamp electrophysiological studies confirmed that the observed Kv1.3 upregulation translated to increased Kv1.3 channel activity. The kinase Fyn, a risk factor for PD, modulated transcriptional upregulation and posttranslational modification of microglial Kv1.3. Multiple state-of-the-art analyses, including Duolink proximity ligation assay imaging, revealed that Fyn directly bound to Kv1.3 and posttranslationally modified its channel activity. Furthermore, we demonstrated the functional relevance of Kv1.3 in augmenting the neuroinflammatory response by using Kv1.3-KO primary microglia and the Kv1.3-specific small-molecule inhibitor PAP-1, thus highlighting the importance of Kv1.3 in neuroinflammation. Administration of PAP-1 significantly inhibited neurodegeneration and neuroinflammation in multiple animal models of PD. Collectively, our results imply that Fyn-dependent regulation of Kv1.3 channels plays an obligatory role in accentuating the neuroinflammatory response in PD and identify Kv1.3 as a potential therapeutic target for PD.

Authors

Souvarish Sarkar, Hai M. Nguyen, Emir Malovic, Jie Luo, Monica Langley, Bharathi N. Palanisamy, Neeraj Singh, Sireesha Manne, Matthew Neal, Michelle Gabrielle, Ahmed Abdalla, Poojya Anantharam, Dharmin Rokad, Nikhil Panicker, Vikrant Singh, Muhammet Ay, Adhithiya Charli, Dilshan Harischandra, Lee-Way Jin, Huajun Jin, Srikant Rangaraju, Vellareddy Anantharam, Heike Wulff, Anumantha G. Kanthasamy

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Figure 6

Kv1.3 modulates neuroinflammation in a cell culture model of PD.

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Kv1.3 modulates neuroinflammation in a cell culture model of PD. (A–C) K...
(A–C) Kv1.3 WT and KO PMCs were treated with 1 μM αSynAgg for 24 hours. Luminex analysis shows that Kv1.3 KO reduced the release of the αSynAgg-induced proinflammatory factors (A) TNF-α, (B) IL-12, and (C) IL-1β. (D–H) Immortalized MMCs were transfected with WT a Kv1.3 plasmid, and then 48 hours after transfection, cells were treated with 1 μM αSynAgg for 24 hours. (D–F) qRT-PCR analysis showing that Kv1.3 overexpression aggravated αSynAgg-induced production of the proinflammatory factors (D) Nos2, (E) pro–IL-1β, and (F) TNF-α. (G and H) Luminex analysis showing that Kv1.3 overexpression potentiated the release of the proinflammatory factors (G) IL-6 and (H) IL-12. (I) Voltage ramp from –120 mV to 40 mV elicited a characteristic outward rectifying current in αSynAgg-treated microglia that was sensitive to the Kv1.3-selective inhibitor PAP-1. (J) LDH assay showing that PAP-1 reduced αSynAgg-induced LDH release from microglial cells. (K–M) Luminex assay revealing that PAP-1 attenuated the αSynAgg-induced proinflammatory factors (K) IL-12, (L) TNF-α, and (M) IL-6. (N) Western blot analysis demonstrating that PAP-1 reduced αSynAgg-induced NLRP3 expression. (O) ICC analysis revealed that PAP-1 reduced NLRP3 expression induced by αSynAgg. Scale bar: 25 μm. A 1-way ANOVA was performed to compare multiple groups. In most cases, Tukey’s post hoc analysis was applied. Each dot on the bar graphs represents a biological replicate. Data are presented as the mean ± SEM, with 3–4 biological replicates from 2–3 independent experiments. *P ≤ 0.05, **P < 0.01, and ***P < 0.001.