A systems pharmacology-based approach to identify novel Kv1.3 channel-dependent mechanisms in microglial activation

Rangaraju, Srikant; Raza, Syed Ali; Pennati, Andrea; Deng, Qiudong; Dammer, Eric B.; Duong, Duc M.; Pennington, Michael W.; Tansey, Malú G.; Lah, James J.; Betarbet, Ranjita; Seyfried, Nicholas T.; Levey, Aĺlan I.

doi:10.1186/s12974-017-0906-6

Cited by 55 publications

(76 citation statements)

References 64 publications

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“…Although published (Charolidi et al 2015; Nguyen et al 2017; Rangaraju et al 2015; Rangaraju et al 2017; Rus et al 2005) and present data clearly showed that in the brain, Kv1.3 is mainly expressed in activated microglia, it has been described in neurons, most prominently in mitral cells of the olfactory bulb (Fadool et al 2004) and presynaptic terminals of brain stem auditory neurons (Gazula et al 2010), and also in cortical interneurons and in oligodendrocyte progenitor cells but at low levels (Vautier et al 2004; Duque et al 2013), raising the possibility that the above in vivo effects of Kv1.3 knockout may be secondary to Kv1.3 hypomorph in neurons. To address this possibility, we used a chemical biology approach, as in neurons Kv1.3 is typically part of hetereomultimers with Kv1.1, Kv1.2 and Kv1.6 subunits (Helms et al 1997) and therefore has a different pharmacology from the Kv1.3 homotetramers or Kv1.3/Kv1.5 heterotetramers found in T cells, microglia and macrophages (Chandy et al 2004; Feske et al 2015; Wulff et al 2009).…”

Section: Resultsmentioning

confidence: 99%

“…A first in vivo suggestion of such a role of Kv1.3 came from a recent report showing that systemic inflammation induced by intravenous administration of LPS to adult mice resulted in increased ShK-F6CA (a fluorescein-conjugated Kv1.3-specific peptide inhibitor) labeling of microglia and infiltrating macrophages in the brain (Rangaraju et al 2017). In the current study, we stereotactically injected LPS into the ventricles (Maezawa et al 2006).…”

Section: Discussionmentioning

confidence: 99%

“…We previously demonstrated that cultured primary microglia activated by lipopolysaccharides (LPS), which induces a classic M1-like activation state, exhibited high Kv1.3 current densities but virtually no activities of the other major microglial K + channel, the inward-rectifier Kir2.1 (Nguyen et al 2017). A systems pharmacology-based study identified functional roles for Kv1.3 in pro-inflammatory microglial activation (Rangaraju et al 2017) while electrophysiological studies by our group demonstrated upregulated Kv1.3 expression in microglia acutely isolated from the infarct areas of mice subjected to experimental stroke (Chen et al 2016). Kv1.3 blockers have further been demonstrated to inhibit microglia mediated neurotoxicity in culture (Fordyce et al 2005) and to protect mice from microglia mediated radiation-induced brain injury (Peng et al 2014).…”

Section: Introductionmentioning

confidence: 99%

“…Although Kv1.3 appears to be a suitable target for microglia-targeted modulation, a definite in vivo demonstration of the role of Kv1.3 in neuroinflammation is missing, as nearly all prior studies were conducted using BV2 cells, primary microglia, or brain slices in culture (Charolidi et al 2015; Fordyce et al 2005; Khanna et al 2001; Kotecha and Schlichter 1999; Küst et al 1999; Nguyen et al 2017; Rangaraju et al 2017; Schilling and Eder 2003; Schilling and Eder 2011). It is now realized that BV2 cells significantly differ from cultured or in vivo microglia in many aspects (Butovsky et al 2014; Henn et al 2009; Kettenmann et al 2011).…”

Section: Introductionmentioning

confidence: 99%

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The voltage‐gated potassium channel Kv1.3 is required for microglial pro‐inflammatory activation in vivo

Lucente

Nguyen

Wulff

et al. 2018

Glia

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Microglia show a rich repertoire of activation patterns regulated by a complex ensemble of surface ion channels, receptors, and transporters. We and others have investigated whether microglia vary their K channel expression as a means to achieve functional diversity. However, most of the prior studies were conducted using in vitro models such as BV2 cells, primary microglia, or brain slices in culture, which may not accurately reflect microglia physiology in adult individuals. Here we employed an in vivo mouse model of selective innate immune activation by intracerebroventricular injection of lipopolysaccharides (ICV-LPS) to determine the role of the voltage-gated Kv1.3 channel in LPS-induced M1-like microglial activation. Using microglia acutely isolated from adult brains, we detected Kv1.3 and Kir2.1 currents, and found that ICV-LPS increased the current density and RNA expression of Kv1.3 but did not affect those of Kir2.1. Genetic knockout of Kv1.3 abolished LPS-induced microglial activation exemplified by Iba-1 immunoreactivity and expression of pro-inflammatory mediators such as IL-1β, TNF-α, IL-6, and iNOS. Moreover, Kv1.3 knockout mitigated the LPS-induced impairment of hippocampal long-term potentiation (hLTP), suggesting that Kv1.3 activity regulates pro-inflammatory microglial neurotoxicity. Pharmacological intervention using PAP-1, a small molecule that selectively blocks homotetrameric Kv1.3 channels, achieved anti-inflammatory and hLTP-recovery effects similar to Kv1.3 knockout. We conclude that Kv1.3 is required for microglial M1-like pro-inflammatory activation in vivo. A significant implication of our in vivo data is that Kv1.3 blockers could be therapeutic candidates for neurological diseases where microglia-mediated neurotoxicity is implicated in the pathogenesis.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The voltage‐gated potassium channel Kv1.3 is required for microglial pro‐inflammatory activation in vivo

Lucente

Nguyen

Wulff

et al. 2018

Glia

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“…These findings suggest that Kv1.3 inhibition does not affect phagocytosis in vivo confirming a recent study reporting that Kv1.3 blockers to not impair the ability of isolated brain mononuclear cells to phagocytose polystyrene microspheres. 38…”

Section: Kv13 Blockade Preferentially Reduces Inflammatory Cytokine mentioning

confidence: 99%

Inhibition of the potassium channel Kv1.3 reduces infarction and inflammation in ischemic stroke

Chen

Nguyen

Maezawa

et al. 2017

Ann Clin Transl Neurol

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ObjectiveInhibitors of the voltage‐gated K+ channel Kv1.3 are currently in development as immunomodulators for the treatment of autoimmune diseases. As Kv1.3 is also expressed on microglia and has been shown to be specifically up‐regulated on “M1‐like” microglia, we here tested the therapeutic hypothesis that the brain‐penetrant small‐molecule Kv1.3‐inhibitor PAP‐1 reduces secondary inflammatory damage after ischemia/reperfusion.MethodsWe studied microglial Kv1.3 expression using electrophysiology and immunohistochemistry, and evaluated PAP‐1 in hypoxia‐exposed organotypic hippocampal slices and in middle cerebral artery occlusion (MCAO) with 8 days of reperfusion in both adult male C57BL/6J mice (60 min MCAO) and adult male Wistar rats (90 min MCAO). In both models, PAP‐1 administration was started 12 h after reperfusion.ResultsWe observed Kv1.3 staining on activated microglia in ischemic infarcts in mice, rats, and humans and found higher Kv1.3 current densities in acutely isolated microglia from the infarcted hemisphere than in microglia isolated from the contralateral hemisphere of MCAO mice. PAP‐1 reduced microglia activation and increased neuronal survival in hypoxia‐exposed hippocampal slices as effectively as minocycline. In mouse MCAO, PAP‐1 dose‐dependently reduced infarct area, improved neurological deficit score, and reduced brain levels of IL‐1β and IFN‐γ without affecting IL‐10 and brain‐derived nerve growth factor (BDNF) levels or inhibiting ongoing phagocytosis. The beneficial effects on infarct area and neurological deficit score were reproduced in rats providing confirmation in a second species.InterpretationOur findings suggest that Kv1.3 constitutes a promising therapeutic target for preferentially inhibiting “M1‐like” inflammatory microglia/macrophage functions in ischemic stroke.

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Spinal voltage‐gated potassium channel Kv1.3 contributes to neuropathic pain via the promotion of microglial M1 polarization and activation of the NLRP3 inflammasome

Yuan

Han

Manyande

et al. 2022

European Journal of Pain

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Background: Studies have shown that activation of microglia is the main mechanism of neuropathic pain. Kv1.3 channel is a novel therapeutic target for treating neuroin ammatory disorders due to its crucial role in subsets of microglial cells. As such, it may be involved in the processes of neuropathic pain, however, whether Kv1.3 plays a role in neuroin ammation following peripheral nerve injury is unclear.Methods: The spared nerve injury model (SNI) was used to establish neuropathic pain. Western blot and immuno uorescence were used to examine the effect of Kv1.3 in the SNI rats. PAP-1, a Kv1.3 speci c blocker was administered to alleviate neuropathic pain in the SNI rats.Results: Neuropathic pain and allodynia occurred after SNI, the levels of M1 (CD68, iNos) and M2 (CD206, Arg-1) phenotypes were up-regulated in the spinal cord, and the protein levels of NLRP3, caspase-1 and IL-1β were also increased. Pharmacological blocking of Kv1.3 with PAP-1 alleviated hyperpathia induced by SNI. Meanwhile, intrathecal injection of PAP-1 reduced M1 polarization and decreased NLRP3, caspase-1, and IL-1β expressions of protein levels.Conclusion: Our research indicates that the Kv1.3 channel in the spinal cord contributes to neuropathic pain by promoting microglial M1 polarization and activating the NLRP3 in ammasome.

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A systems pharmacology-based approach to identify novel Kv1.3 channel-dependent mechanisms in microglial activation

Cited by 55 publications

References 64 publications

The voltage‐gated potassium channel Kv1.3 is required for microglial pro‐inflammatory activation in vivo

The voltage‐gated potassium channel Kv1.3 is required for microglial pro‐inflammatory activation in vivo

Inhibition of the potassium channel Kv1.3 reduces infarction and inflammation in ischemic stroke

Spinal voltage‐gated potassium channel Kv1.3 contributes to neuropathic pain via the promotion of microglial M1 polarization and activation of the NLRP3 inflammasome

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