Metformin treatment reduces motor and neuropsychiatric phenotypes in the zQ175 mouse model of Huntington disease
Ana Sanchis, María Adelaida García-Gimeno, Antonio José Cañada-Martínez, María Dolores Sequedo, José María Millán, Pascual Sanz, Rafael P Vázquez-Manrique, Ana Sanchis, María Adelaida García-Gimeno, Antonio José Cañada-Martínez, María Dolores Sequedo, José María Millán, Pascual Sanz, Rafael P Vázquez-Manrique
Abstract
Huntington disease is a neurodegenerative condition for which there is no cure to date. Activation of AMP-activated protein kinase has previously been shown to be beneficial in in vitro and in vivo models of Huntington's disease. Moreover, a recent cross-sectional study demonstrated that treatment with metformin, a well-known activator of this enzyme, is associated with better cognitive scores in patients with this disease. We performed a preclinical study using metformin to treat phenotypes of the zQ175 mouse model of Huntington disease. We evaluated behavior (motor and neuropsychiatric function) and molecular phenotypes (aggregation of mutant huntingtin, levels of brain-derived neurotrophic factor, neuronal inflammation, etc.). We also used two models of polyglutamine toxicity in Caenorhabditis elegans to further explore potential mechanisms of metformin action. Our results provide strong evidence that metformin alleviates motor and neuropsychiatric phenotypes in zQ175 mice. Moreover, metformin intake reduces the number of nuclear aggregates of mutant huntingtin in the striatum. The expression of brain-derived neurotrophic factor, which is reduced in mutant animals, is partially restored in metformin-treated mice, and glial activation in mutant mice is reduced in metformin-treated animals. In addition, using worm models of polyglutamine toxicity, we demonstrate that metformin reduces polyglutamine aggregates and restores neuronal function through mechanisms involving AMP-activated protein kinase and lysosomal function. Our data indicate that metformin alleviates the progression of the disease and further supports AMP-activated protein kinase as a druggable target against Huntington's disease.
Conflict of interest statement
The authors declare that they have no conflict of interest.
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References
- Krobitsch S, Kazantsev AG. Huntington's disease: from molecular basis to therapeutic advances. Int. J. Biochem. Cell Biol. 2011;43:20–24. doi: 10.1016/j.biocel.2010.10.014.
- Ferrante RJ, Kowall NW, Richardson EP., Jr Proliferative and degenerative changes in striatal spiny neurons in Huntington's disease: a combined study using the section-Golgi method and calbindin D28k immunocytochemistry. J. Neurosci. 1991;11:3877–3887. doi: 10.1523/JNEUROSCI.11-12-03877.1991.
- Yamamoto A, Cremona ML, Rothman JE. Autophagy-mediated clearance of huntingtin aggregates triggered by the insulin-signaling pathway. J. Cell Biol. 2006;172:719–731. doi: 10.1083/jcb.200510065.
- Ortega Z, Lucas JJ. Ubiquitin-proteasome system involvement in Huntington’s disease. Front. Mol. Neurosci. 2014;7:77. doi: 10.3389/fnmol.2014.00077.
- Herrero-Martin G, et al. TAK1 activates AMPK-dependent cytoprotective autophagy in TRAIL-treated epithelial cells. EMBO J. 2009;28:677–685. doi: 10.1038/emboj.2009.8.
- Liang J, et al. The energy sensing LKB1-AMPK pathway regulatesp27(kip1) phosphorylation mediating the decision to enter autophagy or apoptosis. Nat. Cell Biol. 2007;9:218–224. doi: 10.1038/ncb1537.
- Carling D, Thornton C, Woods A, Sanders MJ. AMP-activated protein kinase: new regulation, new roles? Biochem. J. 2012;445:11–27. doi: 10.1042/BJ20120546.
- Hardie DG, Ashford ML. AMPK: regulating energy balance at the cellular and whole body levels. Physiology. 2014;29:99–107. doi: 10.1152/physiol.00050.2013.
- Vazquez-Manrique RP, et al. AMPK activation protects from neuronal dysfunction and vulnerability across nematode, cellular and mouse models of Huntington's disease. Hum. Mol. Genet. 2015;25:1043–1058. doi: 10.1093/hmg/ddv513.
- Harper SQ, et al. RNA interference improves motor and neuropathological abnormalities in a Huntington's disease mouse model. Proc. Natl. Acad. Sci. USA. 2005;102:5820–5825. doi: 10.1073/pnas.0501507102.
- Yamamoto A, Lucas JJ, Hen R. Reversal of neuropathology and motor dysfunction in a conditional model of Huntington's disease. Cell. 2000;101:57–66. doi: 10.1016/S0092-8674(00)80623-6.
- Shaw RJ, et al. The kinase LKB1 mediates glucose homeostasis in liver and therapeutic effects of metformin. Science. 2005;310:1642–1646. doi: 10.1126/science.1120781.
- Zhou G, et al. Role of AMP-activated protein kinase in mechanism of metformin action. J. Clin. Invest. 2001;108:1167–1174. doi: 10.1172/JCI13505.
- Witters LA. The blooming of the French lilac. J. Clin. Invest. 2001;108:1105–1107. doi: 10.1172/JCI14178.
- Yokoyama NN, et al. When anti-aging studies meet cancer chemoprevention: can anti-aging agent kill two birds with one blow? Curr. Pharm. Rep. 2015;1:420–433. doi: 10.1007/s40495-015-0039-5.
- Nasri H, Rafieian-Kopaei M. Metformin and diabetic kidney disease: a mini-review on recent findings. Iran J. Pediatr. 2014;24:565–568.
- Ma TC, et al. Metformin therapy in a transgenic mouse model of Huntington’s disease. Neurosci. Lett. 2007;411:98–103. doi: 10.1016/j.neulet.2006.10.039.
- Hervás D, et al. Metformin intake associates with better cognitive function in patients with Huntington's disease. PLoS ONE. 2017;12:e0179283. doi: 10.1371/journal.pone.0179283.
- Lewis JA, Fleming JT. Basic culture methods. Methods Cell Biol. 1995;48:3–29. doi: 10.1016/S0091-679X(08)61381-3.
- Mello C, Fire A. DNA transformation. Methods Cell Biol. 1995;48:451–482. doi: 10.1016/S0091-679X(08)61399-0.
- Walker DS, et al. Inositol 1,4,5-trisphosphate signalling regulates the avoidance response to nose touch in Caenorhabditis elegans. PLoS Genet. 2009;5:e1000636. doi: 10.1371/journal.pgen.1000636.
- Frokjaer-Jensen C, et al. Single-copy insertion of transgenes in Caenorhabditis elegans. Nat. Genet. 2008;40:1375–1383. doi: 10.1038/ng.248.
- Parker JA, et al. Expanded polyglutamines in Caenorhabditis elegans cause axonal abnormalities and severe dysfunction of PLM mechanosensory neurons without cell death. Proc. Natl. Acad. Sci. USA. 2001;98:13318–13323. doi: 10.1073/pnas.231476398.
- Vazquez-Manrique RP, et al. The frataxin-encoding operon of Caenorhabditis elegans shows complex structure and regulation. Genomics. 2007;89:392–401. doi: 10.1016/j.ygeno.2006.10.007.
- Vazquez-Manrique RP, Legg JC, Olofsson B, Ly S, Baylis HA. Improved gene targeting in C. elegans using counter-selection and Flp-mediated marker excision. Genomics. 2010;95:37–46. doi: 10.1016/j.ygeno.2009.09.001.
- Foretz M, Guigas B, Bertrand L, Pollak M, Viollet B. Metformin: from mechanisms of action to therapies. Cell Metab. 2014;20:953–966. doi: 10.1016/j.cmet.2014.09.018.
- Can, A. et al. The tail suspension test. J. Vis. Exp.59, e3769 (2012).
- Brooks SP, Dunnett SB. Tests to assess motor phenotype in mice: a user's guide. Nat. Rev. Neurosci. 2009;10:519–529. doi: 10.1038/nrn2652.
- Schindelin J, et al. Fiji: an open-source platform for biological-image analysis. Nat. Methods. 2012;9:676–682. doi: 10.1038/nmeth.2019.
- Ruifrok AC, Johnston DA. Quantification of histochemical staining by color deconvolution. Anal. Quant. Cytol. Histol. 2001;23:291–299.
- Gibson-Corley KN, Olivier AK, Meyerholz DK. Principles for valid histopathologic scoring in research. Vet. Pathol. 2013;50:1007–1015. doi: 10.1177/0300985813485099.
- Morley JF, Brignull HR, Weyers JJ, Morimoto RI. The threshold for polyglutamine-expansion protein aggregation and cellular toxicity is dynamic and influenced by aging in Caenorhabditis elegans. Proc. Natl. Acad. Sci. USA. 2002;99:10417–10422. doi: 10.1073/pnas.152161099.
- Klionsky DJ, et al. Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy. 2012;8:445–544. doi: 10.4161/auto.19496.
- Menalled LB, et al. Comprehensive behavioral and molecular characterization of a new knock-in mouse model of Huntington’s disease: zQ175. PLoS ONE. 2012;7:e49838. doi: 10.1371/journal.pone.0049838.
- Chermat R, Thierry B, Mico JA, Steru L, Simon P. Adaptation of the tail suspension test to the rat. J. Pharm. 1986;17:348–350.
- Steru L, et al. The automated Tail Suspension Test: a computerized device which differentiates psychotropic drugs. Prog. Neuropsychopharmacol. Biol. Psychiatry. 1987;11:659–671. doi: 10.1016/0278-5846(87)90002-9.
- Steru L, Chermat R, Thierry B, Simon P. The tail suspension test: a new method for screening antidepressants in mice. Psychopharmacology (Berl.) 1985;85:367–370. doi: 10.1007/BF00428203.
- Cryan JF, Mombereau C. In search of a depressed mouse: utility of models for studying depression-related behavior in genetically modified mice. Mol. Psychiatry. 2004;9:326–357. doi: 10.1038/sj.mp.4001457.
- Labuzek K, et al. Quantification of metformin by the HPLC method in brain regions, cerebrospinal fluid and plasma of rats treated with lipopolysaccharide. Pharm. Rep. 2010;62:956–965. doi: 10.1016/S1734-1140(10)70357-1.
- Han I, You Y, Kordower JH, Brady ST, Morfini GA. Differential vulnerability of neurons in Huntington’s disease: the role of cell type-specific features. J. Neurochem. 2010;113:1073–1091.
- Saudou F, Finkbeiner S, Devys D, Greenberg ME. Huntingtin acts in the nucleus to induce apoptosis but death does not correlate with the formation of intranuclear inclusions. Cell. 1998;95:55–66. doi: 10.1016/S0092-8674(00)81782-1.
- Ferrer I, Goutan E, Marin C, Rey MJ, Ribalta T. Brain-derived neurotrophic factor in Huntington disease. Brain Res. 2000;866:257–261. doi: 10.1016/S0006-8993(00)02237-X.
- Zuccato C, et al. Loss of huntingtin-mediated BDNF gene transcription in Huntington's disease. Science. 2001;293:493–498. doi: 10.1126/science.1059581.
- Zuccato C, et al. Systematic assessment of BDNF and its receptor levels in human cortices affected by Huntington's disease. Brain Pathol. 2008;18:225–238. doi: 10.1111/j.1750-3639.2007.00111.x.
- Ma Q, Yang J, Li T, Milner TA, Hempstead BL. Selective reduction of striatal mature BDNF without induction of proBDNF in the zQ175 mouse model of Huntington's disease. Neurobiol. Dis. 2015;82:466–477. doi: 10.1016/j.nbd.2015.08.008.
- Zuccato C, Valenza M, Cattaneo E. Molecular mechanisms and potential therapeutical targets in Huntington’s disease. Physiol. Rev. 2010;90:905–981. doi: 10.1152/physrev.00041.2009.
- Jansen AH, et al. Frequency of nuclear mutant huntingtin inclusion formation in neurons and glia is cell-type-specific. Glia. 2017;65:50–61. doi: 10.1002/glia.23050.
- Crotti A, Glass CK. The choreography of neuroinflammation in Huntington’s disease. Trends Immunol. 2015;36:364–373. doi: 10.1016/j.it.2015.04.007.
- Peng Q, et al. Characterization of behavioral, neuropathological, brain metabolic and key molecular changes in zQ175 knock-in mouse model of Huntington’s disease. PLoS ONE. 2016;11:e0148839. doi: 10.1371/journal.pone.0148839.
- Imai Y, Ibata I, Ito D, Ohsawa K, Kohsaka S. A novel gene iba1 in the major histocompatibility complex class III region encoding an EF hand protein expressed in a monocytic lineage. Biochem. Biophys. Res. Commun. 1996;224:855–862. doi: 10.1006/bbrc.1996.1112.
- Simmons DA, et al. Ferritin accumulation in dystrophic microglia is an early event in the development of Huntington's disease. Glia. 2007;55:1074–1084. doi: 10.1002/glia.20526.
- Bowles KR, Jones L. Kinase signalling in Huntington’s disease. J. Huntingt. Dis. 2014;3:89–123.
- Winder WW, Hardie DG. AMP-activated protein kinase, a metabolic master switch: possible roles in type 2 diabetes. Am. J. Physiol. 1999;277:E1–E10.
- Markowicz-Piasecka M, et al. Metformin—a Future Therapy For Neurodegenerative Diseases. Pharm. Res. 2017;34:2614–2627. doi: 10.1007/s11095-017-2199-y.
- Heikkinen T, et al. Characterization of neurophysiological and behavioral changes, MRI brain volumetry and 1H MRS in zQ175 knock-in mouse model of Huntington's disease. PLoS ONE. 2012;7:e50717. doi: 10.1371/journal.pone.0050717.
- Southwell AL, et al. An enhanced Q175 knock-in mouse model of Huntington disease with higher mutant huntingtin levels and accelerated disease phenotypes. Hum. Mol. Genet. 2016;25:3654–3675. doi: 10.1093/hmg/ddw212.
- Carty N, et al. Characterization of HTT inclusion size, location, and timing in the zQ175 mouse model of Huntington's disease: an in vivo high-content imaging study. PLoS ONE. 2015;10:e0123527. doi: 10.1371/journal.pone.0123527.
- Arnoux I, et al. Metformin reverses early cortical network dysfunction and behavior changes in Huntington’s disease. Elife. 2018;7:e38744. doi: 10.7554/eLife.38744.
- Lin SC, Hardie DG. AMPK: sensing glucose as well as cellular energy status. Cell Metab. 2018;27:299–313. doi: 10.1016/j.cmet.2017.10.009.
- Giampa C, et al. Systemic delivery of recombinant brain derived neurotrophic factor (BDNF) in the R6/2 mouse model of Huntington’s disease. PLoS ONE. 2013;8:e64037. doi: 10.1371/journal.pone.0064037.
- Brenner S. The genetics of Caenorhabditis elegans. Genetics. 1974;77:71–94.
- Apfeld J, O'Connor G, McDonagh T, DiStefano PS, Curtis R. The AMP-activated protein kinase AAK-2 links energy levels and insulin-like signals to lifespan in C. elegans. Genes Dev. 2004;18:3004–3009. doi: 10.1101/gad.1255404.
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