A SCN10A SNP biases human pain sensitivity

Guangyou Duan, Chongyang Han, Qingli Wang, Shanna Guo, Yuhao Zhang, Ying Ying, Penghao Huang, Li Zhang, Lawrence Macala, Palak Shah, Mi Zhang, Ningbo Li, Sulayman D Dib-Hajj, Stephen G Waxman, Xianwei Zhang, Guangyou Duan, Chongyang Han, Qingli Wang, Shanna Guo, Yuhao Zhang, Ying Ying, Penghao Huang, Li Zhang, Lawrence Macala, Palak Shah, Mi Zhang, Ningbo Li, Sulayman D Dib-Hajj, Stephen G Waxman, Xianwei Zhang

Abstract

Background: Nav1.8 sodium channels, encoded by SCN10A, are preferentially expressed in nociceptive neurons and play an important role in human pain. Although rare gain-of-function variants in SCN10A have been identified in individuals with painful peripheral neuropathies, whether more common variants in SCN10A can have an effect at the channel level and at the dorsal root ganglion, neuronal level leading to a pain disorder or an altered normal pain threshold has not been determined.

Results: Candidate single nucleotide polymorphism association approach together with experimental pain testing in human subjects was used to explore possible common SCN10A missense variants that might affect human pain sensitivity. We demonstrated an association between rs6795970 (G > A; p.Ala1073Val) and higher thresholds for mechanical pain in a discovery cohort (496 subjects) and confirmed it in a larger replication cohort (1005 female subjects). Functional assessments showed that although the minor allele shifts channel activation by -4.3 mV, a proexcitatory attribute, it accelerates inactivation, an antiexcitatory attribute, with the net effect being reduced repetitive firing of dorsal root ganglion neurons, consistent with lower mechanical pain sensitivity.

Conclusions: At the association and mechanistic levels, the SCN10A single nucleotide polymorphism rs6795970 biases human pain sensitivity.

Keywords: Nav1.8; dorsal root ganglion; pain; voltage-gated sodium channel.

© The Author(s) 2016.

Figures

Figure 1.
Figure 1.
Schematic of sodium channel polypeptide showing the locations of the corresponded amino acid residues encoded by the five SCN10A SNPs.
Figure 2.
Figure 2.
Mean mechanical pain measurement values for the different genotypes of rs6795970 in replication study. (a) dull pressure pain threshold (D-PPT) and dull pressure pain tolerance (D-PTO); (b) sharp pressure pain threshold (S-PPT) and sharp pressure pain tolerance (S-PTO); (c) quantizing pricking pain (QPT). Compared to A/A, *Bonferroni significance of difference P < 0.05; **P < 0.01; ***P < 0.001.
Figure 3.
Figure 3.
Voltage-clamp analysis of Nav1.8-Ala1073 and Nav1.8-Val1073 channels. (a and b) Representative Nav1.8 current family traces recorded from mouse Nav1.8-knockout DRG neurons transfected with Nav1.8-Ala1073 (a) or Nav1.8-Val1073 (b) channels. Cells were held at −70 mV and stepped to a range of potentials (−70 to +40 mV in 5-mV increments) for 100 ms. (c) Comparison of voltage-dependent activation between Nav1.8-Ala1073 and Nav1.8-Val1073 channels. Activation of Nav1.8-Val1073 channels is hyperpolarized by 4.3 mV compared with Nav1.8-Ala1073 channels. (d) Fast inactivation of Nav1.8-Val1073 channels is not significantly different from that of Nav1.8-Ala1073 channels. (e) Slow inactivation of Nav1.8-Val1073 channels is not significantly different from that of Nav1.8-Ala1073 channels. (f) Nav1.8-Val1073 channels display faster inactivation kinetics compared with Nav1.8-Ala1073 channels, *P < 0.05. (g) Comparison of persistent current between Nav1.8-Ala1073 channels and Nav1.8-Val1073 channels. (h) Comparison of recovery from fast inactivation between Nav1.8-Ala1073 and Nav1.8-Val1073 channels at −70 mV and −50 mV. Nav1.8-Val1073 channels tend to recovery more slowly from inactivation than Nav1.8-Ala1073 channels at both −70 mV and −50 mV.
Figure 4.
Figure 4.
Compared with Nav1.8-Ala1073 channels, Nav1.8-Val1073 channels decrease firing frequency in small DRG neurons. (a) Response of cells expressing Nav1.8-Ala1073 and Nav1.8-Val1073 channels, respectively, to 500-ms depolarizing current steps that are 1X, 2X, and 3X (left, middle, and right traces, respectively) the current threshold for action potential generation. (b) Comparison of mean fire frequencies among cells expressing Nav1.8-Ala1073 and Nav1.8-Val1073 channels across the range of current injections from 50 to 500 pA, *P < 0.05, indicating statistical significance between two groups of neurons.

References

    1. Akopian AN, Sivilotti L, Wood JN. A tetrodotoxin-resistant voltage-gated sodium channel expressed by sensory neurons. Nature 1996; 379: 257–262.
    1. Black JA, Waxman SG. Molecular identities of two tetrodotoxin-resistant sodium channels in corneal axons. Exp Eye Res 2002; 75: 193–199.
    1. Persson AK, Black JA, Gasser A, et al. Sodium-calcium exchanger and multiple sodium channel isoforms in intra-epidermal nerve terminals. Mol Pain 2010; 6: 84.
    1. Sangameswaran L, Fish LM, Koch BD, et al. A novel tetrodotoxin-sensitive, voltage-gated sodium channel expressed in rat and human dorsal root ganglia. J Biol Chem 1997; 272: 14805–14809.
    1. Bennett DL, Woods CG. Painful and painless channelopathies. Lancet Neurol 2014; 13: 587–599.
    1. Dib-Hajj SD, Cummins TR, Black JA, et al. Sodium channels in normal and pathological pain. Annu Rev Neurosci 2010; 33: 325–347.
    1. Garrison SR, Weyer AD, Barabas ME, et al. A gain-of-function voltage-gated sodium channel 1.8 mutation drives intense hyperexcitability of A- and C-fiber neurons. Pain 2014; 155: 896–905.
    1. Renganathan M, Cummins TR, Waxman SG. Contribution of Na(v)1.8 sodium channels to action potential electrogenesis in DRG neurons. J Neurophysiol 2001; 86: 629–640.
    1. Faber CG, Lauria G, Merkies IS, et al. Gain-of-function Nav1.8 mutations in painful neuropathy. Proc Natl Acad Sci USA 2012; 109: 19444–19449.
    1. Han C, Vasylyev D, Macala LJ, et al. The G1662S NaV1.8 mutation in small fibre neuropathy: impaired inactivation underlying DRG neuron hyperexcitability. J Neurol Neurosurg Psychiatry 2014; 85: 499–505.
    1. Huang J, Yang Y, Zhao P, et al. Small-fiber neuropathy Nav1.8 mutation shifts activation to hyperpolarized potentials and increases excitability of dorsal root ganglion neurons. J Neurosci 2013; 33: 14087–14097.
    1. Bezzina CR, Barc J, Mizusawa Y, et al. Common variants at SCN5A-SCN10A and HEY2 are associated with Brugada syndrome, a rare disease with high risk of sudden cardiac death. Nat Genet 2013; 45: 1044–1049.
    1. Chambers JC, Zhao J, Terracciano CM, et al. Genetic variation in SCN10A influences cardiac conduction. Nat Genet 2010; 42: 149–152.
    1. Denny JC, Ritchie MD, Crawford DC, et al. Identification of genomic predictors of atrioventricular conduction: using electronic medical records as a tool for genome science. Circulation 2010; 122: 2016–2021.
    1. Holm H, Gudbjartsson DF, Arnar DO, et al. Several common variants modulate heart rate, PR interval and QRS duration. Nat Genet 2010; 42: 117–122.
    1. Jeff JM, Ritchie MD, Denny JC, et al. Generalization of variants identified by genome-wide association studies for electrocardiographic traits in African Americans. Ann Hum Genet 2013; 77: 321–332.
    1. Pfeufer A, van Noord C, Marciante KD, et al. Genome-wide association study of PR interval. Nat Genet 2010; 42: 153–159.
    1. Ritchie MD, Denny JC, Zuvich RL, et al. Genome- and phenome-wide analyses of cardiac conduction identifies markers of arrhythmia risk. Circulation 2013; 127: 1377–1385.
    1. Sano M, Kamitsuji S, Kamatani N, et al. Genome-wide association study of electrocardiographic parameters identifies a new association for PR interval and confirms previously reported associations. Hum Mol Genet 2014; 23: 6668–6676.
    1. Smith JG, Magnani JW, Palmer C, et al. Genome-wide association studies of the PR interval in African Americans. PLoS Genet 2011; 7: e1001304.
    1. Sotoodehnia N, Isaacs A, de Bakker PI, et al. Common variants in 22 loci are associated with QRS duration and cardiac ventricular conduction. Nat Genet 2010; 42: 1068–1076.
    1. Behr ER, Savio-Galimberti E, Barc J, et al. Role of common and rare variants in SCN10A: results from the Brugada syndrome QRS locus gene discovery collaborative study. Cardiovasc Res 2015; 106: 520–529.
    1. Jabbari J, Olesen MS, Yuan L, et al. Common and rare variants in SCN10A modulate the risk of atrial fibrillation. Circ Cardiovasc Genet 2015; 8: 64–73.
    1. Delaney JT, Muhammad R, Shi Y, et al. Common SCN10A variants modulate PR interval and heart rate response during atrial fibrillation. Europace 2014; 16: 485–490.
    1. Verkerk AO, Remme CA, Schumacher CA, et al. Functional Nav1.8 channels in intracardiac neurons: the link between SCN10A and cardiac electrophysiology. Circ Res 2012; 111: 333–343.
    1. Yang T, Atack TC, Stroud DM, et al. Blocking Scn10a channels in heart reduces late sodium current and is antiarrhythmic. Circ Res 2012; 111: 322–332.
    1. Duan G, Guo S, Zhang Y, et al. The effect of SCN9A variation on basal pain sensitivity in the general population: an experimental study in young women. J Pain 2015; 16: 971–980.
    1. Pauli P, Wiedemann G, Nickola M. Pressure pain thresholds asymmetry in left- and right-handers: associations with behavioural measures of cerebral laterality. Eur J Pain 1999; 3: 151–156.
    1. Chanock SJ, Manolio T, Boehnke M, et al. Replicating genotype-phenotype associations. Nature 2007; 447: 655–660.
    1. Tian C, Gregersen PK, Seldin MF. Accounting for ancestry: population substructure and genome-wide association studies. Hum Mol Genet 2008; 17: R143–150.
    1. Duan G, Guo S, Zhan H, et al. A new real-time method for detecting the effect of fentanyl using the preoperative pressure pain threshold and Narcotrend index: a randomized study in female surgery patients. Medicine (Baltimore) 2015; 94: e316.
    1. Guo S, Duan G, Wang J, et al. Comparison of sufentanil-tramadol PCIA between laparoscopic cholecystectomy and gynecological laparoscopy. Zhong hua Wai Ke Za Zhi 2015; 53: 150–154.
    1. Fillingim RB, King CD, Ribeiro-Dasilva MC, et al. Sex, gender, and pain: a review of recent clinical and experimental findings. J Pain 2009; 10: 447–485.
    1. Duan G, Xiang G, Zhang X, et al. An improvement of mechanical pain sensitivity measurement method: the smaller sized probes may detect heterogeneous sensory threshold in healthy male subjects. Pain Med 2014; 15: 272–280.
    1. Duan G, Xiang G, Zhang X, et al. A single-nucleotide polymorphism in SCN9A may decrease postoperative pain sensitivity in the general population. Anesthesiology 2013; 118: 436–442.
    1. Montagne-Clavel J, Oliveras JL. The “plantar test” apparatus (Ugo Basile Biological Apparatus), a controlled infrared noxious radiant heat stimulus for precise withdrawal latency measurement in the rat, as a tool for humans? Somatosens Mot Res 1996; 13: 215–223.
    1. Sinville R, Coyne J, Meagher RJ, et al. Ligase detection reaction for the analysis of point mutations using free-solution conjugate electrophoresis in a polymer microfluidic device. Electrophoresis 2008; 29: 4751–4760.
    1. Atkins JF, Wills NM, Loughran G, et al. A case for “StopGo”: reprogramming translation to augment codon meaning of GGN by promoting unconventional termination (Stop) after addition of glycine and then allowing continued translation (Go). RNA 2007; 13: 803–810.
    1. Luke GA, de Felipe P, Lukashev A, et al. Occurrence, function and evolutionary origins of ‘2 A- like’ sequences in virus genomes. J Gen Virol 2008; 89: 1036–1042.
    1. Ryan MD, Drew J. Foot-and-mouth disease virus 2 A oligopeptide mediated cleavage of an artificial polyprotein. EMBO J 1994; 13: 928–933.
    1. Dib-Hajj SD, Choi JS, Macala LJ, et al. Transfection of rat or mouse neurons by biolistics or electroporation. Nat Protoc 2009; 4: 1118–1126.
    1. Stirling LC, Forlani G, Baker MD, et al. Nociceptor-specific gene deletion using heterozygous NaV1.8-Cre recombinase mice. Pain 2005; 113: 27–36.
    1. Amir R, Michaelis M, Devor M. Membrane potential oscillations in dorsal root ganglion neurons: role in normal electrogenesis and neuropathic pain. J Neurosci 1999; 19: 8589–8596.
    1. Amir R, Michaelis M, Devor M. Burst discharge in primary sensory neurons: triggered by subthreshold oscillations, maintained by depolarizing afterpotentials. J Neurosci 2002; 22: 1187–1198.
    1. Castillo RC, Wegener ST, Heins SE, et al. Longitudinal relationships between anxiety, depression, and pain: results from a two-year cohort study of lower extremity trauma patients. Pain 2013; 154: 2860–2866.
    1. Kroenke K, Wu J, Bair MJ, et al. Reciprocal relationship between pain and depression: a 12-month longitudinal analysis in primary care. J Pain 2011; 12: 964–973.
    1. Riley JL, 3rd, Cruz-Almeida Y, Glover TL, et al. Age and race effects on pain sensitivity and modulation among middle-aged and older adults. J Pain 2014; 15: 272–282.
    1. Estacion M, Harty TP, Choi JS, et al. A sodium channel gene SCN9A polymorphism that increases nociceptor excitability. Ann Neurol 2009; 66: 862–866.
    1. Huang J, Yang Y, Dib-Hajj SD, et al. Depolarized inactivation overcomes impaired activation to produce DRG neuron hyperexcitability in a Nav1.7 mutation in a patient with distal limb pain. J Neurosci 2014; 34: 12328–12340.
    1. Dib-Hajj SD, Yang Y, Black JA, et al. The Na(V)1.7 sodium channel: from molecule to man. Nat Rev Neurosci 2013; 14: 49–62.
    1. Faber CG, Hoeijmakers JG, Ahn HS, et al. Gain of function Nanu1.7 mutations in idiopathic small fiber neuropathy. Ann Neurol 2012; 71: 26–39.
    1. Fertleman CR, Baker MD, Parker KA, et al. SCN9A mutations in paroxysmal extreme pain disorder: allelic variants underlie distinct channel defects and phenotypes. Neuron 2006; 52: 767–774.
    1. Reimann F, Cox JJ, Belfer I, et al. Pain perception is altered by a nucleotide polymorphism in SCN9A. Proc Natl Acad Sci U S A 2010; 107: 5148–5153.

Source: PubMed

Sponsorit ja yhteistyökumppanit

Sairaudet

Huumeiden interventiot

3
Tilaa