Gene therapy for neurological diseases requires efficient gene delivery to target tissues in the central and peripheral nervous systems. Although adeno-associated virus is one of the most promising vectors for clinical use against neurological diseases, it is difficult to get it across the blood–brain barrier. A clinically practical approach to using a vector based on adeno-associated virus to decrease the expression of a specific gene in both the central and the peripheral nervous system has yet to be established. Here, we analyzed whether upper lumbar intrathecal administration of a therapeutic vector incorporating adeno-associated virus and short-hairpin RNA against superoxide dismutase-1 bypassed the blood–brain barrier to target the spinal cord and dorsal root ganglia. The therapeutic vector effectively suppressed mRNA and protein expression of endogenous superoxide dismutase-1 in the lumbar spinal cord and dorsal root ganglia. Moreover, neither neurological side effects nor toxicity due to the incorporated short-hairpin RNA occurred after the injection. We propose that this approach could be developed into novel therapies for motor neuron diseases and chronic pain conditions, such as complex regional pain syndrome, through silencing of the genes responsible for pathologies in the spinal cord and dorsal root ganglia.
Hirai and colleagues demonstrate that intrathecal injection of AAV9 vector encoding short hairpin RNA against SOD1 reduces the mRNA expression levels of the targeted molecule by approximately 60% in both the spinal cord and the dorsal root ganglia of adult mice, and that this reduction persists for at least 4 weeks.
Get full access to this article
View all access options for this article.
References
1.
AlexanderG.M., ErwinK.L., ByersN.et al.2004. Effect of transgene copy number on survival in the G93A SOD1 transgenic mouse model of ALS. Brain Res. Mol. Brain Res., 130:7–15.
2.
BevanA.K., DuqueS., FoustK.D.et al.2011. Systemic gene delivery in large species for targeting spinal cord, brain, and peripheral tissues for pediatric disorders. Mol. Ther., 19:1971–1980.
3.
BishL.T., MorineK., SleeperM.M.et al.2008. Adeno-associated virus (AAV) serotype 9 provides global cardiac gene transfer superior to AAV1, AAV6, AAV7, and AAV8 in the mouse and rat. Hum. Gene Ther., 19:1359–1368.
4.
ChristophT., GrünwellerA., MikaJ.et al.2006. Silencing of vanilloid receptor TRPV1 by RNAi reduces neuropathic and visceral pain in vivo. Biochem. Biophys. Res. Commun., 350:238–243.
5.
ChristophT., BahrenbergG., De VryJ.et al.2008. Investigation of TRPV1 loss-of-function phenotypes in transgenic shRNA expressing and knockout mice. Mol. Cell. Neurosci., 37:579–589.
FedericiT., TaubJ.S., BaumG.R.et al.2011. Robust spinal motor neuron transduction following intrathecal delivery of AAV9 in pigs. Gene Ther., 15. 10.1038/gt.2011.130.
8.
FireA., XuS., MontgomeryM.K.et al.1998. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature, 391:806–811.
9.
FuH., MuenzerJ., SamulskiR.J.et al.2003. Self-complementary adeno-associated virus serotype 2 vector: Global distribution and broad dispersion of AAV-mediated transgene expression in mouse brain. Mol. Ther., 8:911–917.
10.
FuH., DirosarioJ., KilledarS.et al.2011. Correction of neurological disease of mucopolysaccharidosis IIIB in adult mice by rAAV9 trans-blood–brain barrier gene delivery. Mol. Ther., 19:1025–1033.
11.
FuY., ZhangQ., KangC.et al.2009. Inhibitory effects of adenovirus mediated COX-2, Akt1 and PIK3R1 shRNA on the growth of malignant tumor cells in vitro and in vivo. Int. J. Oncol., 35:583–591.
12.
GarrawayS.M., XuQ., InturrisiC.E.2009. siRNA-mediated knockdown of the NR1 subunit gene of the NMDA receptor attenuates formalin-induced pain behaviors in adult rats. J. Pain, 10:380–390.
13.
GregorevicP., BlankinshipM.J., AllenJ.M.et al.2004. Systemic delivery of genes to striated muscles using adeno-associated viral vectors. Nat. Med., 10:828–834.
14.
GrimmD., StreetzK.L., JoplingC.L.et al.2006. Fatality in mice due to oversaturation of cellular microRNA/short hairpin RNA pathways. Nature, 441:537–541.
15.
HargreavesK., DubnerR., BrownF.et al.1988. A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia. Pain, 32:77–88.
16.
HassaniZ., LemkineG.F., ErbacherP.et al.2005. Lipid-mediated siRNA delivery down-regulates exogenous gene expression in the mouse brain at picomolar levels. J. Gene Med., 7:198–207.
17.
HermensW.T., Ter BrakeO., DijkhuizenP.A.et al.1999. Purification of recombinant adeno-associated virus by iodixanol gradient ultracentrifugation allows rapid and reproducible preparation of vector stocks for gene transfer in the nervous system. Hum. Gene Ther., 10:1885–1891.
18.
JansonC., McPheeS., BilaniukL.et al.2002. Clinical protocol: Gene therapy of Canavan disease: AAV-2 vector for neurosurgical delivery of aspartoacylase gene (ASPA) to the human brain. Hum. Gene Ther., 13:1391–1412.
19.
KaemmererW.F., ReddyR.G., WarlickC.A.et al.2000. In vivo transduction of cerebellar Purkinje cells using adeno-associated virus vectors. Mol. Ther., 2:446–457.
20.
KappelS., MatthessY., KaufmannM., StrebhardtK.2007. Silencing of mammalian genes by tetracycline-inducible shRNA expression. Nat. Protoc., 2:3257–3269.
21.
KawaseM., MurakamiK., FujimuraM.et al.1999. Exacerbation of delayed cell injury after transient global ischemia in mutant mice with CuZn superoxide dismutase deficiency. Stroke, 30:1962–1968.
22.
LiG., LiD., XieQ.et al.2008. RNA interfering connective tissue growth factor prevents rat hepatic stellate cell activation and extracellular matrix production. J. Gene Med., 10:1039–1047.
23.
LuoM.C., ZhangD.Q., MaS.W.et al.2005. An efficient intrathecal delivery of small interfering RNA to the spinal cord and peripheral neurons. Mol. Pain, 1:29.
24.
MannoC.S., PierceG.F., ArrudaV.R.et al.2006. Successful transduction of liver in hemophilia by AAV-Factor IX and limitations imposed by the host immune response. Nat. Med., 12:342–347.
25.
MarksW.J., OstremJ.L., VerhagenL.et al.2008. Safety and tolerability of intraputaminal delivery of CERE-120 (adeno-associated virus serotype 2-neurturin) to patients with idiopathic Parkinson's disease: An open-label, phase I trial. Lancet Neurol., 7:400–408.
26.
MayraA., TomimitsuH., KuboderaT.et al.2011. Intraperitoneal AAV9-shRNA inhibits target expression in neonatal skeletal and cardiac muscles. Biochem. Biophys. Res. Commun., 405:204–209.
27.
MikamiM., YangJ.2005. Short hairpin RNA-mediated selective knockdown of NaV1.8 tetrodotoxin-resistant voltage-gated sodium channel in dorsal root ganglion neurons. Anesthesiology, 103:828–836.
28.
MitchellM., JerebtsovaM., BatshawM.L.et al.2000. Long-term gene transfer to mouse fetuses with recombinant adenovirus and adeno-associated virus (AAV) vectors. Gene Ther., 7:1986–1992.
29.
PanH.L., ZhangY.Q., ZhaoZ.Q.2010. Involvement of lysophosphatidic acid in bone cancer pain by potentiation of TRPV1 via PKCɛ pathway in dorsal root ganglion neurons. Mol. Pain, 6:85.
30.
PavlakisP.P., AlexopoulosH., KosmidisM.L.et al.2011. Peripheral neuropathies in Sjögren syndrome: A new reappraisal. J. Neurol. Neurosurg. Psychiatry, 82:798–802.
31.
PedenC.S., BurgerC., MuzyczkaN., MandelR.J.2004. Circulating anti-wild-type adeno-associated virus type 2 (AAV2) antibodies inhibit recombinant AAV2 (rAAV2)-mediated, but not rAAV5-mediated, gene transfer in the brain. J. Virol., 78:6344–6359.
32.
PogatzkiE.M., ZahnP.K., BrennanT.J.2000. Lumbar catheterization of the subarachnoid space with a 32-gauge polyurethane catheter in the rat. Eur. J. Pain, 4:111–113.
33.
PuskovicV., WolfeD., GossJ.et al.2004. Prolonged biologically active transgene expression driven by HSV LAP2 in brain in vivo. Mol. Ther., 10:67–75.
34.
ReaumeA.G., ElliottJ.L., HoffmanE.K.et al.1996. Motor neurons in Cu/Zn superoxide dismutase-deficient mice develop normally but exhibit enhanced cell death after axonal injury. Nat. Genet., 13:43–47.
35.
Rodriguez-LebronE., Denovan-WrightE.M., NashK.et al.2005. Intrastriatal rAAV-mediated delivery of anti-huntingtin shRNAs induces partial reversal of disease progression in R6/1 Huntington's disease transgenic mice. Mol. Ther., 12:618–633.
36.
RossiJ.J.2008. Expression strategies for short hairpin RNA interference triggers. Hum. Gene Ther., 19:313–317.
37.
SaitoY., YokotaT., MitaniT.et al.2005. Transgenic small interfering RNA halts amyotrophic lateral sclerosis in a mouse model. J. Biol. Chem., 280:42826–42830.
38.
SakuraS., HashimotoK., BollenA.W.et al.1996. Intrathecal catheterization in the rat: Improved technique for morphologic analysis of drug-induced injury. Anesthesiology, 85:1184–1189.
39.
SenechalY., KellyP.H., CryanJ.F.et al.2007. Amyloid precursor protein knockdown by siRNA impairs spontaneous alternation in adult mice. J. Neurochem., 102:1928–1940.
40.
SennC., HangartnerC., MoesS.et al.2005. Central administration of small interfering RNAs in rats: A comparison with antisense oligonucleotides. Eur. J. Pharmacol., 522:30–37.
41.
SnyderB.R., GrayS.J., QuachE.T.et al.2011. Comparison of adeno-associated viral vector serotypes for spinal cord and motor neuron gene delivery. Hum. Gene Ther., 22:1129–1135.
42.
StorekB., ReinhardtM., WangC.et al.2008. Sensory neuron targeting by self-complementary AAV8 via lumbar puncture for chronic pain. Proc. Natl. Acad. Sci. U.S.A., 105:1055–1060.
43.
TanP.H., YangL.C., ShihH.C.et al.2005. Gene knockdown with intrathecal siRNA of NMDA receptor NR2B subunit reduces formalin-induced nociception in the rat. Gene Ther., 12:59–66.
44.
TowneC., RaoulC., SchneiderB.L., AebischerP.2008. Systemic AAV6 delivery mediating RNA interference against SOD1: Neuromuscular transduction does not alter disease progression in fALS mice. Mol. Ther., 16:1018–1025.
45.
UnoY., PiaoW., MiyataK.et al.2011. High-density lipoprotein facilitates in vivo delivery of α-tocopherol-conjugated short-interfering RNA to the brain. Hum. Gene Ther., 22:711–719.
46.
WeiT., LiW.W., GuoT.Z.et al.2009. Post-junctional facilitation of substance P signaling in a tibia fracture rat model of complex regional pain syndrome type I. Pain, 144:278–286.
47.
WorgallS., SondhiD., HackettN.R.et al.2008. Treatment of late infantile neuronal ceroid lipofuscinosis by CNS administration of a serotype 2 adeno-associated virus expressing CLN2 cDNA. Hum. Gene Ther., 19:463–474.
48.
YokotaT., MiyagishiM., HinoT.et al.2004. siRNA-based inhibition specific for mutant SOD1 with single nucleotide alternation in familial ALS, compared with ribozyme and DNA enzyme. Biochem. Biophys. Res. Commun., 314:283–291.
49.
YokotaT., SasaguriH., SaitoY.et al.2007. Increase of disease duration of amyotrophic lateral sclerosis in a mouse model by transgenic small interfering RNA. Arch. Neurol., 64:145–146.
