Differential Modulation of the Voltage-Gated Na+ Channel 1.6 by Peptides Derived From Fibroblast Growth Factor 14

Singh, A.K.; Dvorak, Nolan M.; Tapia, Cynthia M.; Mosebarger, Angela; Ali, Shariq; Bullock, Zaniqua; Chen, Haiying; Zhou, Jia; Laezza, Fernanda

doi:10.3389/fmolb.2021.742903

Cited by 11 publications

(16 citation statements)

References 68 publications

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“…Consistent with the results of previous investigations [14,[25][26][27][28]30], co-expression of FGF14 with the Na v 1.6 channel in heterologous cells lead to a reduction in the peak I Na density of Na v 1.6-mediated currents, as evidenced by HEK-Na v 1.6-FGF14-GFP cells displaying an average peak I Na density (−17.31 ± 2.5 pA/pF; n = 10) significantly less than HEK-Na v 1.6-GFP cells (−57.83 ± 6.3 pA/pF; n = 8; Figure 4A-C). Whereas Wee1 inhibitor II exacerbated this FGF14-mediated regulatory on Na v 1.2-mediated currents (Figure 2A-C), pharmacological inhibition of Wee1 kinase did not affect Na v 1.6-mediated peak I Na density in the absence or presence of FGF14 (Figure 4A-C).…”

Section: Wee1 Inhibitor II Does Not Affect Fgf14-mediated Regulation Of the Na V 16 Channelsupporting

confidence: 93%

“…Whereas Wee1 inhibitor II exacerbated this FGF14-mediated regulatory on Na v 1.2-mediated currents (Figure 2A-C), pharmacological inhibition of Wee1 kinase did not affect Na v 1.6-mediated peak I Na density in the absence or presence of FGF14 (Figure 4A-C). Consistent with previous investigations [14,[25][26][27][28]30], co-expression of FGF14 with the Na v 1.6 channel lead to a slowing of Na v 1.6 channel fast inactivation (Figure 4D) and a depolarizing shift in the voltage-dependence of Na v 1.6 channel activation (Figure 4E,F). These findings are evidenced by HEK-Na v 1.6-GFP cells displaying a tau of fast inactivation of 1.03 ± 0.04 ms (n = 8), whereas HEK-Na v 1.6-FGF14-GFP cells display a significantly larger value for this parameter of 1.64 ± 0.26 ms (n = 8; Figure 4D), and HEK-Na v 1.6-GFP cells displaying a V 1/2 of activation of −22.87 ± 1.69 mV (n = 8), whereas HEK-Na v 1.6-FGF14-GFP cells display a significantly more depolarized V 1/2 of activation of −18.15 ± 1.03 mV (n = 8; Figure 4E,F), respectively.…”

Section: Wee1 Inhibitor II Does Not Affect Fgf14-mediated Regulation Of the Na V 16 Channelsupporting

confidence: 91%

“…The following plasmid constructs used in this study were engineered and characterized as previously described [8,14,15,[23][24][25][26][27][28]: CLuc-FGF14, CD4-Na v 1.2 CTD-NLuc, CD4-Na v 1.6 CTD-NLuc, GFP, FGF14-GFP, and FGF14 Y158A -GFP.…”

Section: Plasmid Constructsmentioning

confidence: 99%

“…Clampex 9.2 software (Molecular Devices) was used to set experimental parameters, and electrophysiological equipment was interfaced to this software using a Digidata 1200 analog-digital interface (Molecular Devices). Acquired data was then analyzed as previously described [27,28,30,31,33].…”

Section: Whole-cell Voltage-clamp Recordings In Heterologous Cellsmentioning

confidence: 99%

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Pharmacological Inhibition of Wee1 Kinase Selectively Modulates the Voltage-Gated Na+ Channel 1.2 Macromolecular Complex

Dvorak

Tapia

Baumgartner

et al. 2021

Cells

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Voltage-gated Na+ (Nav) channels are a primary molecular determinant of the action potential (AP). Despite the canonical role of the pore-forming α subunit in conferring this function, protein–protein interactions (PPI) between the Nav channel α subunit and its auxiliary proteins are necessary to reconstitute the full physiological activity of the channel and to fine-tune neuronal excitability. In the brain, the Nav channel isoforms 1.2 (Nav1.2) and 1.6 (Nav1.6) are enriched, and their activities are differentially regulated by the Nav channel auxiliary protein fibroblast growth factor 14 (FGF14). Despite the known regulation of neuronal Nav channel activity by FGF14, less is known about cellular signaling molecules that might modulate these regulatory effects of FGF14. To that end, and building upon our previous investigations suggesting that neuronal Nav channel activity is regulated by a kinase network involving GSK3, AKT, and Wee1, we interrogate in our current investigation how pharmacological inhibition of Wee1 kinase, a serine/threonine and tyrosine kinase that is a crucial component of the G2-M cell cycle checkpoint, affects the Nav1.2 and Nav1.6 channel macromolecular complexes. Our results show that the highly selective inhibitor of Wee1 kinase, called Wee1 inhibitor II, modulates FGF14:Nav1.2 complex assembly, but does not significantly affect FGF14:Nav1.6 complex assembly. These results are functionally recapitulated, as Wee1 inhibitor II entirely alters FGF14-mediated regulation of the Nav1.2 channel, but displays no effects on the Nav1.6 channel. At the molecular level, these effects of Wee1 inhibitor II on FGF14:Nav1.2 complex assembly and FGF14-mediated regulation of Nav1.2-mediated Na+ currents are shown to be dependent upon the presence of Y158 of FGF14, a residue known to be a prominent site for phosphorylation-mediated regulation of the protein. Overall, our data suggest that pharmacological inhibition of Wee1 confers selective modulatory effects on Nav1.2 channel activity, which has important implications for unraveling cellular signaling pathways that fine-tune neuronal excitability.

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Section: Wee1 Inhibitor II Does Not Affect Fgf14-mediated Regulation Of the Na V 16 Channelsupporting

confidence: 93%

Section: Wee1 Inhibitor II Does Not Affect Fgf14-mediated Regulation Of the Na V 16 Channelsupporting

confidence: 91%

Section: Plasmid Constructsmentioning

confidence: 99%

Section: Whole-cell Voltage-clamp Recordings In Heterologous Cellsmentioning

confidence: 99%

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Pharmacological Inhibition of Wee1 Kinase Selectively Modulates the Voltage-Gated Na+ Channel 1.2 Macromolecular Complex

Dvorak

Tapia

Baumgartner

et al. 2021

Cells

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“…However, the opposite effects of FGF14, the protein from which PW201 is derived, in heterologous cells versus neurons is widely recognized [12,17,24]. Specifically, co-expression of FGF14 with the Na v 1.6 channel in heterologous systems has previously been shown to suppress Na v 1.6-mediated I Na [12,15,24,38,40,41,60,61], whereas over-expression of FGF14 in neurons has been shown to increase I Na [17]. As such, these opposite effects observed for PW201 in heterologous cells versus in neurons are unsurprising and provide supporting evidence for the compound functioning as a partial pharmacological mimic of FGF14.…”

Section: Discussionmentioning

confidence: 99%

Pharmacologically Targeting the Fibroblast Growth Factor 14 Interaction Site on the Voltage-Gated Na+ Channel 1.6 Enables Isoform-Selective Modulation

Dvorak

Tapia

Singh

et al. 2021

IJMS

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Voltage-gated Na+ (Nav) channels are the primary molecular determinant of the action potential. Among the nine isoforms of the Nav channel α subunit that have been described (Nav1.1-Nav1.9), Nav1.1, Nav1.2, and Nav1.6 are the primary isoforms expressed in the central nervous system (CNS). Crucially, these three CNS Nav channel isoforms display differential expression across neuronal cell types and diverge with respect to their subcellular distributions. Considering these differences in terms of their localization, the CNS Nav channel isoforms could represent promising targets for the development of targeted neuromodulators. However, current therapeutics that target Nav channels lack selectivity, which results in deleterious side effects due to modulation of off-target Nav channel isoforms. Among the structural components of the Nav channel α subunit that could be pharmacologically targeted to achieve isoform selectivity, the C-terminal domains (CTD) of Nav channels represent promising candidates on account of displaying appreciable amino acid sequence divergence that enables functionally unique protein–protein interactions (PPIs) with Nav channel auxiliary proteins. In medium spiny neurons (MSNs) of the nucleus accumbens (NAc), a critical brain region of the mesocorticolimbic circuit, the PPI between the CTD of the Nav1.6 channel and its auxiliary protein fibroblast growth factor 14 (FGF14) is central to the generation of electrical outputs, underscoring its potential value as a site for targeted neuromodulation. Focusing on this PPI, we previously developed a peptidomimetic derived from residues of FGF14 that have an interaction site on the CTD of the Nav1.6 channel. In this work, we show that whereas the compound displays dose-dependent effects on the activity of Nav1.6 channels in heterologous cells, the compound does not affect Nav1.1 or Nav1.2 channels at comparable concentrations. In addition, we show that the compound correspondingly modulates the action potential discharge and the transient Na+ of MSNs of the NAc. Overall, these results demonstrate that pharmacologically targeting the FGF14 interaction site on the CTD of the Nav1.6 channel is a strategy to achieve isoform-selective modulation, and, more broadly, that sites on the CTDs of Nav channels interacted with by auxiliary proteins could represent candidates for the development of targeted therapeutics.

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New developments in the biology of fibroblast growth factors

Ornitz

Itoh

2022

WIREs Mechanisms of Disease

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The fibroblast growth factor (FGF) family is composed of 18 secreted signaling proteins consisting of canonical FGFs and endocrine FGFs that activate four receptor tyrosine kinases (FGFRs 1-4) and four intracellular proteins (intracellular FGFs or iFGFs) that primarily function to regulate the activity of voltagegated sodium channels and other molecules. The canonical FGFs, endocrine FGFs, and iFGFs have been reviewed extensively by us and others. In this review, we briefly summarize past reviews and then focus on new developments in the FGF field since our last review in 2015. Some of the highlights in the past 6 years include the use of optogenetic tools, viral vectors, and inducible transgenes to experimentally modulate FGF signaling, the clinical use of small molecule FGFR inhibitors, an expanded understanding of endocrine FGF signaling, functions for FGF signaling in stem cell pluripotency and differentiation, roles for FGF signaling in tissue homeostasis and regeneration, a continuing elaboration of mechanisms of FGF signaling in development, and an expanding appreciation of roles for FGF signaling in neuropsychiatric diseases.

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Differential Modulation of the Voltage-Gated Na+ Channel 1.6 by Peptides Derived From Fibroblast Growth Factor 14

Cited by 11 publications

References 68 publications

Pharmacological Inhibition of Wee1 Kinase Selectively Modulates the Voltage-Gated Na+ Channel 1.2 Macromolecular Complex

Pharmacological Inhibition of Wee1 Kinase Selectively Modulates the Voltage-Gated Na+ Channel 1.2 Macromolecular Complex

Pharmacologically Targeting the Fibroblast Growth Factor 14 Interaction Site on the Voltage-Gated Na+ Channel 1.6 Enables Isoform-Selective Modulation

New developments in the biology of fibroblast growth factors

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