Lentiviral Overexpression of GRK6 Alleviates l-Dopa–Induced Dyskinesia in Experimental Parkinson’s Disease
Science Translational Medicine • 21 Apr 2010 • Vol 2, Issue 28 • p. 28ra28 • DOI: 10.1126/scitranslmed.3000664
Treatment for Tremors Without Side Effects
As neurodegenerative diseases go, Parkinson’s disease is fairly treatable. Oral doses of l-dopa can still the tremors and normalize a patient’s movements—for a time. Eventually, however, most patients develop involuntary aimless gestures call dyskinesias, thought to be a result of oversensitive dopamine responses in the brain, caused by years of taking l-dopa. Now, Bezard and his colleagues have taken aim at a regulator of the dopamine receptor, G protein–coupled receptor kinase 6 (GRK6), to combat these disturbing side effects.
The dopamine receptor, like others in its family, will desensitize after use. In this state, the receptor can no longer be activated and is taken up by the cell. The first step in desensitization is the phosphorylation of the receptor by GRK6. After many years of l-dopa, the amount of GRK in the brain starts to decline and the machinery that desensitizes the receptor does not work properly, leading, it is believed, to the uncontrolled movements of dyskinesia. The authors reinstated GRKs with gene therapy in mice that had an induced parkinsonian syndrome and showed that the dyskinesia-like movements of the mice were much reduced and, as expected, desensitization of the dopamine receptor was normalized. Repeating this experiment in macaque monkeys, in which a Parkinson-like disease had been artificially induced by a toxic agent, gave similar results: Increasing GRK6 expression in the brain could markedly improve the dyskinesia-like side effects of long-term l-dopa treatment, likely by correcting the desensitization of dopamine receptors. Notably, correction of GRK6 did not interfere with the therapeutic effects of l-dopa—an important attribute for the eventual application of such a therapy.
These authors have identified a signaling pathway that seems to be responsible for the worst side effect of the standard treatment for Parkinson’s disease. Manipulation of one of its members, GRK6, or other components of dopamine receptor sensitization may prove to be an effective treatment for these side effects without hindering the efficacy of one of the most useful drugs in the neurologist’s armamentarium.
Abstract
Parkinson’s disease is caused primarily by degeneration of brain dopaminergic neurons in the substantia nigra and the consequent deficit of dopamine in the striatum. Dopamine replacement therapy with the dopamine precursor l-dopa is the mainstay of current treatment. After several years, however, the patients develop l-dopa–induced dyskinesia, or abnormal involuntary movements, thought to be due to excessive signaling via dopamine receptors. G protein–coupled receptor kinases (GRKs) control desensitization of dopamine receptors. We found that dyskinesia is attenuated by lentivirus-mediated overexpression of GRK6 in the striatum in rodent and primate models of Parkinson’s disease. Conversely, reduction of GRK6 concentration by microRNA delivered with lentiviral vector exacerbated dyskinesia in parkinsonian rats. GRK6 suppressed dyskinesia in monkeys without compromising the antiparkinsonian effects of l-dopa and even prolonged the antiparkinsonian effect of a lower dose of l-dopa. Our finding that increased availability of GRK6 ameliorates dyskinesia and increases duration of the antiparkinsonian action of l-dopa suggests a promising approach for controlling both dyskinesia and motor fluctuations in Parkinson’s disease.
Get full access to this article
View all available purchase options and get full access to this article.
Already a Subscriber?Sign In
Supplementary Material
Summary
Materials and Methods
Fig. S1. The GFP-tagged GRK6 is functional and has the subcellular localization of the endogenous GRK6.
Fig. S2. Antibodies to GRK6 selectively recognize GRK6A or GRK6B splicing variants.
Fig. S3. The lentivirus carrying two chained miRNAs targets both GRK6A and GRK6B splice variants.
Fig. S4. Infection of the rat striatum with the miRNA lentivirus induces the GRK6 knockdown.
References
Resources
File (2-28ra28_sm.pdf)
References and Notes
1
Fahn S., How do you treat motor complications in Parkinson’s disease: Medicine, surgery, or both? Ann. Neurol. 64 (suppl. 2), S56–S64 (2008).
2
Bezard E., Gross C. E., Qin L., Gurevich V. V., Benovic J. L., Gurevich E. V., L-DOPA reverses the MPTP-induced elevation of the arrestin2 and GRK6 expression and enhanced ERK activation in monkey brain. Neurobiol. Dis. 18, 323–335 (2005).
3
Bychkov E., Ahmed M. R., Dalby K. N., Gurevich E. V., Dopamine depletion and subsequent treatment with l-DOPA, but not the long-lived dopamine agonist pergolide, enhances activity of the Akt pathway in the rat striatum. J. Neurochem. 102, 699–711 (2007).
4
Sgambato-Faure V., Buggia V., Gilbert F., Lévesque D., Benabid A. L., Berger F., Coordinated and spatial upregulation of arc in striatonigral neurons correlates with L-dopa-induced behavioral sensitization in dyskinetic rats. J. Neuropathol. Exp. Neurol. 64, 936–947 (2005).
5
Aubert I., Guigoni C., Håkansson K., Li Q., Dovero S., Barthe N., Bioulac B. H., Gross C. E., Fisone G., Bloch B., Bezard E., Increased D1 dopamine receptor signaling in levodopa-induced dyskinesia. Ann. Neurol. 57, 17–26 (2005).
6
Kovoor A., Seyffarth P., Ebert J., Barghshoon S., Chen C. K., Schwarz S., Axelrod J. D., Cheyette B. N., Simon M. I., Lester H. A., Schwarz J., D2 dopamine receptors colocalize regulator of G-protein signaling 9-2 (RGS9-2) via the RGS9 DEP domain, and RGS9 knock-out mice develop dyskinesias associated with dopamine pathways. J. Neurosci. 25, 2157–2165 (2005).
7
Gold S. J., Hoang C. V., Potts B. W., Porras G., Pioli E., Kim K. W., Nadjar A., Qin C., LaHoste G. J., Li Q., Bioulac B. H., Waugh J. L., Gurevich E., Neve R. L., Bezard E., RGS9–2 negatively modulates l-3,4-dihydroxyphenylalanine-induced dyskinesia in experimental Parkinson’s disease. J. Neurosci. 27, 14338–14348 (2007).
8
Bézard E., Ferry S., Mach U., Stark H., Leriche L., Boraud T., Gross C., Sokoloff P., Attenuation of levodopa-induced dyskinesia by normalizing dopamine D3 receptor function. Nat. Med. 9, 762–767 (2003).
9
DeWire S. M., Ahn S., Lefkowitz R. J., Shenoy S. K., β-Arrestins and cell signaling. Annu. Rev. Physiol. 69, 483–510 (2007).
10
Gurevich E. V., Gurevich V. V., Arrestins: Ubiquitous regulators of cellular signaling pathways. Genome Biol. 7, 236 (2006).
11
Bohn L. M., Gainetdinov R. R., Sotnikova T. D., Medvedev I. O., Lefkowitz R. J., Dykstra L. A., Caron M. G., Enhanced rewarding properties of morphine, but not cocaine, in βarrestin-2 knock-out mice. J. Neurosci. 23, 10265–10273 (2003).
12
Bohn L. M., Lefkowitz R. J., Gainetdinov R. R., Peppel K., Caron M. G., Lin F. T., Enhanced morphine analgesia in mice lacking β-arrestin 2. Science 286, 2495–2498 (1999).
13
Gainetdinov R. R., Bohn L. M., Sotnikova T. D., Cyr M., Laakso A., Macrae A. D., Torres G. E., Kim K. M., Lefkowitz R. J., Caron M. G., Premont R. T., Dopaminergic supersensitivity in G protein-coupled receptor kinase 6-deficient mice. Neuron 38, 291–303 (2003).
14
Gainetdinov R. R., Bohn L. M., Walker J. K., Laporte S. A., Macrae A. D., Caron M. G., Lefkowitz R. J., Premont R. T., Muscarinic supersensitivity and impaired receptor desensitization in G protein–coupled receptor kinase 5–deficient mice. Neuron 24, 1029–1036 (1999).
15
Ahmed M. R., Bychkov E., Gurevich V. V., Benovic J. L., Gurevich E. V., Altered expression and subcellular distribution of GRK subtypes in the dopamine-depleted rat basal ganglia is not normalized by l-DOPA treatment. J. Neurochem. 104, 1622–1636 (2008).
16
Guigoni C., Doudnikoff E., Li Q., Bloch B., Bezard E., Altered D1 dopamine receptor trafficking in parkinsonian and dyskinetic non-human primates. Neurobiol. Dis. 26, 452–463 (2007).
17
Arriza J. L., Dawson T. M., Simerly R. B., Martin L. J., Caron M. G., Snyder S. H., Lefkowitz R. J., The G-protein–coupled receptor kinases βARK1 and βARK2 are widely distributed at synapses in rat brain. J. Neurosci. 12, 4045–4055 (1992).
18
Gurevich E. V., Benovic J. L., Gurevich V. V., Arrestin2 expression selectively increases during neural differentiation. J. Neurochem. 91, 1404–1416 (2004).
19
Bychkov E. R., Gurevich V. V., Joyce J. N., Benovic J. L., Gurevich E. V., Arrestins and two receptor kinases are upregulated in Parkinson’s disease with dementia. Neurobiol. Aging 29, 379–396 (2008).
20
Firsov D., Elalouf J.-M., Molecular cloning of two rat GRK6 splice variants. Am. J. Physiol. 273, C953–C961 (1997).
21
Fienberg A. A., Hiroi N., Mermelstein P. G., Song W., Snyder G. L., Nishi A., Cheramy A., O’Callaghan J. P., Miller D. B., Cole D. G., Corbett R., Haile C. N., Cooper D. C., Onn S. P., Grace A. A., Ouimet C. C., White F. J., Hyman S. E., Surmeier D. J., Girault J., Nestler E. J., Greengard P., DARPP-32: Regulator of the efficacy of dopaminergic neurotransmission. Science 281, 838–842 (1998).
22
Bordet R., Ridray S., Carboni S., Diaz J., Sokoloff P., Schwartz J. C., Induction of dopamine D3 receptor expression as a mechanism of behavioral sensitization to levodopa. Proc. Natl. Acad. Sci. U.S.A. 94, 3363–3367 (1997).
23
Cenci M. A., Lee C. S., Björklund A., L-DOPA-induced dyskinesia in the rat is associated with striatal overexpression of prodynorphin- and glutamic acid decarboxylase mRNA. Eur. J. Neurosci. 10, 2694–2706 (1998).
24
Lundblad M., Andersson M., Winkler C., Kirik D., Wierup N., Cenci M. A., Pharmacological validation of behavioural measures of akinesia and dyskinesia in a rat model of Parkinson’s disease. Eur. J. Neurosci. 15, 120–132 (2002).
25
Berthet A., Porras G., Doudnikoff E., Stark H., Cador M., Bezard E., Bloch B., Pharmacological analysis demonstrates dramatic alteration of D1 dopamine receptor neuronal distribution in the rat analog of l-DOPA-induced dyskinesia. J. Neurosci. 29, 4829–4835 (2009).
26
Gerfen C. R., Miyachi S., Paletzki R., Brown P., D1 dopamine receptor supersensitivity in the dopamine-depleted striatum results from a switch in the regulation of ERK1/2/MAP kinase. J. Neurosci. 22, 5042–5054 (2002).
27
Le Moine C., Bloch B., D1 and D2 dopamine receptor gene expression in the rat striatum: Sensitive cRNA probes demonstrate prominent segregation of D1 and D2 mRNAs in distinct neuronal populations of the dorsal and ventral striatum. J. Comp. Neurol. 355, 418–426 (1995).
28
Morissette M., Grondin R., Goulet M., Bédard P. J., Di Paolo T., Differential regulation of striatal preproenkephalin and preprotachykinin mRNA levels in MPTP-lesioned monkeys chronically treated with dopamine D1 or D2 receptor agonists. J. Neurochem. 72, 682–692 (1999).
29
Bezard E., Brotchie J. M., Gross C. E., Pathophysiology of levodopa-induced dyskinesia: Potential for new therapies. Nat. Rev. Neurosci. 2, 577–588 (2001).
30
Aubert I., Guigoni C., Li Q., Dovero S., Bioulac B. H., Gross C. E., Crossman A. R., Bloch B., Bezard E., Enhanced preproenkephalin-B–derived opioid transmission in striatum and subthalamic nucleus converges upon globus pallidus internalis in L-3,4-dihydroxyphenylalanine–induced dyskinesia. Biol. Psychiatry 61, 836–844 (2007).
31
Guigoni C., Aubert I., Li Q., Gurevich V. V., Benovic J. L., Ferry S., Mach U., Stark H., Leriche L., Håkansson K., Bioulac B. H., Gross C. E., Sokoloff P., Fisone G., Gurevich E. V., Bloch B., Bezard E., Pathogenesis of levodopa-induced dyskinesia: Focus on D1 and D3 dopamine receptors. Parkinsonism Relat. Disord. 11 (suppl. 1), S25–S29 (2005).
32
Hollinger S., Hepler J. R., Cellular regulation of RGS proteins: Modulators and integrators of G protein signaling. Pharmacol. Rev. 54, 527–559 (2002).
33
Moore C. A., Milano S. K., Benovic J. L., Regulation of receptor trafficking by GRKs and arrestins. Annu. Rev. Physiol. 69, 451–482 (2007).
34
Kim K. M., Valenzano K. J., Robinson S. R., Yao W. D., Barak L. S., Caron M. G., Differential regulation of the dopamine D2 and D3 receptors by G protein-coupled receptor kinases and β-arrestins. J. Biol. Chem. 276, 37409–37414 (2001).
35
Tiberi M., Nash S. R., Bertrand L., Lefkowitz R. J., Caron M. G., Differential regulation of dopamine D1A receptor responsiveness by various G protein-coupled receptor kinases. J. Biol. Chem. 271, 3771–3778 (1996).
36
Boraud T., Bezard E., Bioulac B., Gross C. E., Dopamine agonist-induced dyskinesias are correlated to both firing pattern and frequency alterations of pallidal neurones in the MPTP-treated monkey. Brain 124, 546–557 (2001).
37
Bezard E., Dovero S., Prunier C., Ravenscroft P., Chalon S., Guilloteau D., Crossman A. R., Bioulac B., Brotchie J. M., Gross C. E., Relationship between the appearance of symptoms and the level of nigrostriatal degeneration in a progressive 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-lesioned macaque model of Parkinson’s disease. J. Neurosci. 21, 6853–6861 (2001).
38
Bezard E., Boraud T., Chalon S., Brotchie J. M., Guilloteau D., Gross C. E., Pallidal border cells: An anatomical and electrophysiological study in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated monkey. Neuroscience 103, 117–123 (2001).
Information & Authors
Information
Published In
Science Translational Medicine
Volume 2 | Issue 28
April 2010
April 2010
Copyright
Copyright © 2010, American Association for the Advancement of Science.
Submission history
Received: 23 November 2009
Accepted: 2 April 2010
Acknowledgments
Acknowledgments: We thank R. Baishen, J. Li, and H. Li for excellent technical assistance; J.-M. Elalouf and J. L. Benovic for rat and human GRK6 cDNAs; D. Levesque and S. Sabol (NIH, Bethesda, MD) for prodynorphin and preproenkephalin RNA probes; and J. L. Benovic for purified human GRK6A protein. Funding: Agence Nationale de la Recherche, France (E. Bezard); Biothèque Primate–Centre National de la Recherche Scientifique Life Sciences Department (E. Bezard); NIH grants EY011500 (V.V.G.), NS45117 and NS065868 (E.V.G.), and GM077561 and GM081756 (V.V.G.); and Michael J. Fox Foundation for Parkinson's Research (E. Bezard and E.V.G.). Author contributions: E. Bezard and E.V.G. designed and organized the experiments; E.V.G., V.V.G., M.R.A., Y.T.C., and S.K. designed, cloned, and produced viral vectors and viruses; E.V.G., M.R.A., Y.T.C., E. Bychkov, S.K., A.B., and G.P. performed rat behavioral, neurochemical, and histological experiments; E. Bezard, A.B., G.P., Q.L., B.H.B., B.B., I.A., S.D., and E.D. performed monkey behavioral, neurochemical, and histological experiments; E. Bezard and E.V.G. analyzed the data; E. Bezard, V.V.G., and E.V.G. wrote the paper. Competing interests: The authors have declared no competing interests.
Authors
Metrics & Citations
Metrics
Article Usage
Altmetrics
Citations
Export citation
Select the format you want to export the citation of this publication.
Cited by
- Mass spectrometry imaging identifies abnormally elevated brain l-DOPA levels and extrastriatal monoaminergic dysregulation in l-DOPA–induced dyskinesia, Science Advances, 7, 2, (2021)./doi/10.1126/sciadv.abe5948
- Involvement of Autophagy in Levodopa‐Induced Dyskinesia, Movement Disorders, 36, 5, (1137-1146), (2021).https://doi.org/10.1002/mds.28480
- GRK6 Depletion Induces HIF Activity in Lung Adenocarcinoma, Frontiers in Oncology, 11, (2021).https://doi.org/10.3389/fonc.2021.654812
- Identification of distinct pathological signatures induced by patient-derived α-synuclein structures in nonhuman primates, Science Advances, 6, 20, (2020)./doi/10.1126/sciadv.aaz9165
- Use of adeno-associated virus-mediated delivery of mutant huntingtin to study the spreading capacity of the protein in mice and non-human primates, Neurobiology of Disease, 141, (104951), (2020).https://doi.org/10.1016/j.nbd.2020.104951
- Rational Design of a Two‐Photon Fluorogenic Probe for Visualizing Monoamine Oxidase A Activity in Human Glioma Tissues, Angewandte Chemie, 132, 19, (7606-7611), (2020).https://doi.org/10.1002/ange.202000059
- Models of hyperkinetic disorders in primates, Journal of Neuroscience Methods, 332, (108551), (2020).https://doi.org/10.1016/j.jneumeth.2019.108551
- Gastrointestinal and metabolic function in the MPTP-treated macaque model of Parkinson's disease, Heliyon, 6, 12, (e05771), (2020).https://doi.org/10.1016/j.heliyon.2020.e05771
- Rational Design of a Two‐Photon Fluorogenic Probe for Visualizing Monoamine Oxidase A Activity in Human Glioma Tissues, Angewandte Chemie International Edition, 59, 19, (7536-7541), (2020).https://doi.org/10.1002/anie.202000059
- Bidirectional gut-to-brain and brain-to-gut propagation of synucleinopathy in non-human primates, Brain, 143, 5, (1462-1475), (2020).https://doi.org/10.1093/brain/awaa096
- See more
Loading...
View Options
Get Access
Log in to view the full text
AAAS login provides access to Science for AAAS Members, and access to other journals in the Science family to users who have purchased individual subscriptions.
- Become a AAAS Member
- Activate your AAAS ID
- Purchase Access to Other Journals in the Science Family
- Account Help
Log in via OpenAthens.
Log in via Shibboleth.
More options
Register for free to read this article
As a service to the community, this article is available for free. Login or register for free to read this article.
View options
PDF format
Download this article as a PDF file
Download PDF