Huntington disease (HD) is a dominantly inherited neurodegenerative disorder caused by a CAG repeat expansion in the huntingtin gene. We previously discovered that mutant Huntingtin sensitizes type 1 inositol 1,4,5-trisphosphate receptor (InsP3R1) to InsP3. This causes calcium leakage from the endoplasmic reticulum (ER) and a compensatory increase in neuronal store-operated calcium (nSOC) entry. We previously demonstrated that supranormal nSOC leads to synaptic loss in striatal medium spiny neurons (MSNs) in YAC128 HD mice.
Objective:
We sought to identify calcium channels supporting supranormal nSOC in HD MSNs and to validate these channels as potential therapeutic targets for HD.
Methods:
Cortico-striatal cultures were established from wild type and YAC128 HD mice and the density of MSN spines was quantified. The expression of candidate nSOC components was suppressed by RNAi knockdown and by CRISPR/Cas9 knockout. TRPC1 knockout mice were crossed with YAC128 HD mice for evaluation of motor performance in a beamwalk assay.
Results:
RNAi-mediated knockdown of TRPC1, TRPC6, Orai1, or Orai2, but not other TRPC isoforms or Orai3, rescued the density of YAC128 MSN spines. Knockdown of stromal interaction molecule 1 (STIM1), an ER calcium sensor and nSOC activator, also rescued YAC128 MSN spines. Knockdown of the same targets suppressed supranormal nSOC in YAC128 MSN spines. These channel subunits co-immunoprecipitated with STIM1 and STIM2 in synaptosomal lysates from mouse striata. Crossing YAC128 mice with TRPC1 knockout mice improved motor performance and rescued MSN spines in vitro and in vivo, indicating that inhibition of TRPC1 may serve as a neuroprotective strategy for HD treatment.
Conclusions:
TRPC1 channels constitute a potential therapeutic target for treatment of HD.
The Huntington’s Disease Collaborative Research Group. A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington’s disease chromosomes. Cell. 1993;72(6):971–83.
RosasH, KoroshetzW, ChenY, SkeuseC, VangelM, CudkowiczM, et al. Evidence for more widespread cerebral pathology in early HD: An MRI-based morphometric analysis. Neurology. 2003;60(10):1615–20.
4.
ZuccatoC, ValenzaM, CattaneoE. Molecular mechanisms and potential therapeutical targets in Huntington’s disease. Physiol Rev. 2010;90(3):905–81.
5.
MillerBR, BezprozvannyI. Corticostriatal circuit dysfunction in Huntington’s disease: Intersection of glutamate, dopamine, and calcium. Future Neurol. 2010;5:735–56.
6.
BunnerKD, RebecGV. Corticostriatal dysfunction in Huntington’s disease: The basics. Front Hum Neurosci. 2016;10:317.
7.
MilnerwoodAJ, RaymondLA. Early synaptic pathophysiology in neurodegeneration: Insights from Huntington’s disease. Trends Neurosci. 2010;33(11):513–23.
8.
GalvanL, AndreVM, WangEA, CepedaC, LevineMS. Functional differences between direct and indirect striatal output pathways in Huntington’s disease. J Huntingtons Dis. 2012;1(1):17–25.
RyskampD, WuJ, GevaM, KuskoR, GrossmanI, HaydenM, et al. The sigma-1 receptor mediates the beneficial effects of pridopidine in a mouse model of Huntington disease. Neurobiol Dis. 2017;97:46–59.
14.
de YebenesJG, LandwehrmeyerB, SquitieriF, ReilmannR, RosserA, BarkerRA, et al. Pridopidine for the treatment of motor function in patients with Huntington’s disease (MermaiHD): A phase 3, randomised, double-blind, placebo-controlled trial. Lancet Neurol. 2011;10(12):1049–57.
15.
Huntington Study Group HART Investigators. A randomized, double-blind, placebo-controlled trial of pridopidine in Huntington’s disease. Mov Disord. 2013;28(10):1407–15.
16.
KieburtzK, LandwehrmeyerGB, ReilmannR, SavolaJ, EyalE, GrachevI, et al. Efficacy, safety, and tolerability of pridopidine in Huntington disease (HD): Results from the phase II, double-blind, placebo-controlled, dose-ranging study, Pride-HD (P2.005). . Neurology. 2017;88(16 Supplement):P2.005.
17.
McGarryA, KieburtzK, AblerV, GrachevID, GandhiS, AuingerP, et al. Safety and exploratory efficacy at 36 months in Open-HART, an open-label extension study of pridopidine in Huntington’s disease. J Huntingtons Dis. 2017;6(3):189–99.
18.
TangTS, TuH, ChanEY, MaximovA, WangZ, WellingtonCL, et al. Huntingtin and huntingtin-associated protein 1 influence neuronal calcium signaling mediated by inositol-(1,4,5) triphosphate receptor type 1. Neuron. 2003;39(2):227–39.
19.
TangTS, TuH, OrbanPC, ChanEY, HaydenMR, BezprozvannyI. HAP1 facilitates effects of mutant huntingtin on inositol 1,4,5-trisphosphate-induced Ca release in primary culture of striatal medium spiny neurons. Eur J Neurosci. 2004;20(7):1779–87.
20.
TangTS, SlowE, LupuV, StavrovskayaIG, SugimoriM, LlinasR, et al. Disturbed Ca2+ signaling and apoptosis of medium spiny neurons in Huntington’s disease. Proc Natl Acad Sci U S A. 2005;102(7):2602–7.
21.
TangTS, GuoC, WangH, ChenX, BezprozvannyI. Neuroprotective effects of inositol 1,4,5-trisphosphate receptor C-terminal fragment in a Huntington’s disease mouse model. J Neurosci. 2009;29(5):1257–66.
22.
WuJ, RyskampDA, LiangX, EgorovaP, ZakharovaO, HungG, et al. Enhanced store-operated calcium entry leads to striatal synaptic loss in a Huntington’s disease mouse model. J Neurosci. 2016;36(1):125–41.
23.
WuJ, ShihHP, VigontV, HrdlickaL, DigginsL, SinghC, et al. Neuronal store-operated calcium entry pathway as a novel therapeutic target for Huntington’s disease treatment. Chem Biol. 2011;18(6):777–93.
ZengC, TianF, XiaoB. TRPC channels: Prominent candidates of underlying mechanism in neuropsychiatric diseases. Mol Neurobiol. 2016;53(1):631–47.
26.
MocciaF, ZuccoloE, SodaT, TanziF, GuerraG, MapelliL, et al. Stim and Orai proteins in neuronal Ca(2+) signaling and excitability. Front Cell Neurosci. 2015;9:153.
27.
KraftR. STIM and ORAI proteins in the nervous system. Channels. 2015;9(5):245–52.
28.
SelvarajS, SunY, SinghBB. TRPC channels and their implication in neurological diseases. CNS Neurol Disord Drug Targets. 2010;9(1):94–104.
29.
FisherA, BezprozvannyI, WuL, RyskampDA, Bar-NerN, NatanN, et al. AF710B, a novel M1/σ1 agonist with therapeutic efficacy in animal models of Alzheimer’s disease. Neurodegener Dis. 2016;16(1-2):95–110.
30.
LiuX, ChengKT, BandyopadhyayBC, PaniB, DietrichA, PariaBC, et al. Attenuation of store-operated Ca2+ current impairs salivary gland fluid secretion in TRPC1(-/-) mice. Proc Natl Acad Sci U S A. 2007;104(44):17542–7.
31.
SlowEJ, van RaamsdonkJ, RogersD, ColemanSH, GrahamRK, DengY, et al. Selective striatal neuronal loss in a YAC128 mouse model of Huntington disease. Hum Mol Genet. 2003;12(13):1555–67.
32.
SanjanaNE, ShalemO, ZhangF. Improved vectors and genome-wide libraries for CRISPR screening. Nat Methods. 2014;11(8):783–4.
33.
PlattRJ, ChenS, ZhouY, YimMJ, SwiechL, KemptonHR, et al. CRISPR-Cas9 knockin mice for genome editing and cancer modeling. Cell. 2014;159(2):440–55.
34.
SunS, ZhangH, LiuJ, PopugaevaE, XuNJ, FeskeS, et al. Reduced synaptic STIM2 expression and impaired store-operated calcium entry cause destabilization of mature spines in mutant presenilin mice. Neuron. 2014;82(1):79–93.
35.
JiangM, ChenG. High Ca2+-phosphate transfection efficiency in low-density neuronal cultures. Nat Protoc. 2006;1(2):695–700.
36.
TangTS, ChenX, LiuJ, BezprozvannyI. Dopaminergic signaling and striatal neurodegeneration in Huntington’s disease. J Neurosci. 2007;27(30):7899–910.
37.
DumitriuD, RodriguezA, MorrisonJH. High-throughput, detailed, cell-specific neuroanatomy of dendritic spines using microinjection and confocal microscopy. Nat Protoc. 2011;6(9):1391–411.
38.
ZhangH, SunS, WuL, PchitskayaE, ZakharovaO, Fon TacerK, et al. Store-operated calcium channel complex in postsynaptic spines: A new therapeutic target for Alzheimer’s disease treatment. J Neurosci. 2016;36(47):11837–50.
39.
RiccioA, MedhurstAD, MatteiC, KelsellRE, CalverAR, RandallAD, et al. mRNA distribution analysis of human TRPC family in CNS and peripheral tissues. Brain Res Mol Brain Res. 2002;109(1-2):95–104.
40.
ChengKT, OngHL, LiuX, AmbudkarIS. Contribution and regulation of TRPC channels in store-operated Ca2+ entry. Curr Top Membr. 2013;71:149–79.
GwackY, SrikanthS, Oh-HoraM, HoganPG, LampertiED, YamashitaM, et al. Hair loss and defective T-and B-cell function in mice lacking ORAI1. Mol Cell Biol. 2008;28(17):5209–22.
DietrichA, KalwaH, StorchU, y SchnitzlerMM, SalanovaB, PinkenburgO, et al. Pressure-induced and store-operated cation influx in vascular smooth muscle cells is independent of TRPC1. Pflugers Arch. 2007;455(3):465–77.
46.
WangH, ChenX, LiY, TangTS, BezprozvannyI. Tetrabenazine is neuroprotective in Huntington’s disease mice. Mol Neurodegener. 2010;5:18.
47.
LiangX, WuJ, EgorovaP, BezprozvannyI. An automated and quantitative method to evaluate progression of striatal pathology in Huntington’s disease transgenic mice. J Huntingtons Dis. 2014;3(4):343–50.
48.
HeK, QiF, GuoC, ZhanS, XuH, LiuJ, et al. Movement deficits and neuronal loss in basal ganglia in TRPC1 deficient mice. Oncotarget. 2016;7(43):69337.
49.
LuB, FivazM. Neuronal SOCE: Myth or reality?Trends Cell Biol. 2016;26(12):890–3.
50.
MajewskiL, KuznickiJ. SOCE in neurons: Signaling or just refilling?Biochim Biophys Acta. 1853(9):1940–52.
51.
SegalM, KorkotianE. Roles of calcium stores and store-operated channels in plasticity of dendritic spines. Neuroscientist. 2016;22(5):477–85.
52.
PopugaevaE, PchitskayaE, BezprozvannyI. Dysregulation of neuronal calcium homeostasis in Alzheimer’s disease - A therapeutic opportunity?Biochem Biophys Res Commun. 2017;483(4):998–1004.
53.
Berna-ErroA, BraunA, KraftR, KleinschnitzC, SchuhmannMK, StegnerD, et al. STIM2 regulates capacitive Ca2+ entry in neurons and plays a key role in hypoxic neuronal cell death. Sci Signal. 2009;2(93):ra67.
54.
KlejmanME, Gruszczynska-BiegalaJ, Skibinska-KijekA, WisniewskaMB, MisztalK, BlazejczykM, et al. Expression of STIM1 in brain and puncta-like co-localization of STIM1 and ORAI1 upon depletion of Ca(2+) store in neurons. Neurochem Int. 2009;54(1):49–55.
55.
KorkotianE, Oni-BitonE, SegalM. The role of the store-operated calcium entry channel Orai1 in cultured rat hippocampal synapse formation and plasticity. J Physiol. 2017;595(1):125–40.
56.
Gruszczynska-BiegalaJ, PomorskiP, WisniewskaMB, KuznickiJ. Differential roles for STIM1 and STIM2 in store-operated calcium entry in rat neurons. PLoS One. 2011;6(4):e19285.
57.
Gruszczynska-BiegalaJ, KuznickiJ. Native STIM2 and ORAI1 proteins form a calcium-sensitive and thapsigargin-insensitive complex in cortical neurons. J Neurochem. 2013;126(6):727–38.
58.
HartmannJ, KarlRM, AlexanderRP, AdelsbergerH, BrillMS, RuhlmannC, et al. STIM1 controls neuronal Ca(2)(+) signaling, mGluR1-dependent synaptic transmission, and cerebellar motor behavior. Neuron. 2014;82(3):635–44.
59.
StrubingC, KrapivinskyG, KrapivinskyL, ClaphamDE. TRPC1 and TRPC5 form a novel cation channel in mammalian brain. Neuron. 2001;29(3):645–55.
60.
BollimunthaS, SelvarajS, SinghBB. Emerging roles of canonical TRP channels in neuronal function. Adv Exp Med Biol. 2011;704:573–93.
61.
WangY, BuJ, ShenH, LiH, WangZ, ChenG. Targeting transient receptor potential canonical channels for nervous system diseases. Curr Drug Targets. 2017;18(12):1460–5.
62.
VigontV, KolobkovaY, SkopinA, ZiminaO, ZeninV, GlushankovaL, et al. Both Orai1 and TRPC1 are involved in excessive store-operated calcium entry in striatal neurons expressing mutant huntingtin exon 1. Front Physiol. 2015;6:337.
63.
GrigorievI, GouveiaSM, Van der VaartB, DemmersJ, SmythJT, HonnappaS, et al. STIM1 is a MT-plus-end-tracking protein involved in remodeling of the ER. Curr Biol. 2008;18(3):177–82.
64.
HolbroN, GrunditzA, OertnerTG. Differential distribution of endoplasmic reticulum controls metabotropic signaling and plasticity at hippocampal synapses. Proc Natl Acad Sci U S A. 2009;106(35):15055–60.
65.
YuanJP, ZengW, HuangGN, WorleyPF, MuallemS. STIM1 heteromultimerizes TRPC channels to determine their function as store-operated channels. Nat Cell Biol. 2007;9(6):636–45.
66.
OngHL, ChengKT, LiuX, BandyopadhyayBC, PariaBC, SoboloffJ, et al. Dynamic assembly of TRPC1-STIM1-Orai1 ternary complex is involved in store-operated calcium influx. Evidence for similarities in store-operated and calcium release-activated calcium channel components. J Biol Chem. 2007;282(12):9105–16.
67.
NgLC, McCormackMD, AireyJA, SingerCA, KellerPS, ShenXM, et al. TRPC1 and STIM1 mediate capacitative Ca2+ entry in mouse pulmonary arterial smooth muscle cells. J Physiol. 2009;587(11):2429–42.
68.
AlmirzaW, PetersP, van ZoelenE, TheuvenetA. Role of Trpc channels, Stim1 and Orai1 in PGF 2α-induced calcium signaling in NRK fibroblasts. Cell Calcium. 2012;51(1):12–21.
69.
JardinI, GomezLJ, SalidoGM, RosadoJA. Dynamic interaction of hTRPC6 with the Orai1-STIM1 complex or hTRPC3 mediates its role in capacitative or non-capacitative Ca(2+) entry pathways. Biochem J. 2009;420(2):267–76.
70.
NarayananKL, IrmadyK, SubramaniamS, UnsickerK, von Bohlen und HalbachO. Evidence that TRPC1 is involved in hippocampal glutamate-induced cell death. Neurosci Lett. 2008;446(2-3):117–22.
71.
ZhouQ, YenA, RymarczykG, AsaiH, TrengroveC, AzizN, et al. Impairment of PARK14-dependent Ca(2+) signalling is a novel determinant of Parkinson’s disease. Nat Commun. 2016;7:10332.
72.
XingR, ZhangY, XuH, LuoX, ChangRC-C, LiuJ, et al. Spatial memory impairment by TRPC1 depletion is ameliorated by environmental enrichment. Oncotarget. 2016;7(19):27855.
73.
SelvarajS, SunY, WattJA, WangS, LeiS, BirnbaumerL, et al. Neurotoxin-induced ER stress in mouse dopaminergic neurons involves downregulation of TRPC1 and inhibition of AKT/mTOR signaling. J Clin Invest. 2012;122(4):1354–67.
74.
BollimunthaS, SinghBB, ShavaliS, SharmaSK, EbadiM. TRPC1-mediated inhibition of 1-methyl-4-phenylpyridinium ion neurotoxicity in human SH-SY5Y neuroblastoma cells. J Biol Chem. 2005;280(3):2132–40.
SunY, ZhangH, SelvarajS, SukumaranP, LeiS, BirnbaumerL, et al. Inhibition of L-Type Ca2+ channels by TRPC1-STIM1 complex is essential for the protection of dopaminergic neurons. J Neurosci. 2017;37(12):3364–77.
77.
McGurkJS, ShimS, KimJY, WenZ, SongH, MingG-L. Postsynaptic TRPC1 function contributes to BDNF-induced synaptic potentiation at the developing neuromuscular junction. J Neurosci. 2011;31(41):14754–62.
