A rational mechanism for combination treatment of Huntington's disease using lithium and rapamycin.

Huntington's disease (HD) is caused by a polyglutamine expansion mutation in the huntingtin protein that confers a toxic gain-of-function and causes the protein to become aggregate-prone. Aggregate-prone proteins are cleared by macroautophagy, and upregulating this process by rapamycin, which inhibits the mammalian target of rapamycin (mTOR), attenuates their toxicity in various HD models. Recently, we demonstrated that lithium induces mTOR-independent autophagy by inhibiting inositol monophosphatase (IMPase) and reducing inositol and IP3 levels. Here we show that glycogen synthase kinase-3beta (GSK-3beta), another enzyme inhibited by lithium, has opposite effects. In contrast to IMPase inhibition that enhances autophagy, GSK3beta inhibition attenuates autophagy and mutant huntingtin clearance by activating mTOR. In order to counteract the autophagy inhibitory effects of mTOR activation resulting from lithium treatment, we have used the mTOR inhibitor rapamycin in combination with lithium. This combination enhances macroautophagy by mTOR-independent (IMPase inhibition by lithium) and mTOR-dependent (mTOR inhibition by rapamycin) pathways. We provide proof-of-principle for this rational combination treatment approach in vivo by showing greater protection against neurodegeneration in an HD fly model with TOR inhibition and lithium, or in HD flies treated with rapamycin and lithium, compared with either pathway alone.

[1]  K. Lindsten,et al.  Short-lived green fluorescent proteins for quantifying ubiquitin/proteasome-dependent proteolysis in living cells , 2000, Nature Biotechnology.

[2]  Rainer Duden,et al.  Aggregate-prone proteins with polyglutamine and polyalanine expansions are degraded by autophagy. , 2002, Human molecular genetics.

[3]  D. Rubinsztein,et al.  Trehalose, a Novel mTOR-independent Autophagy Enhancer, Accelerates the Clearance of Mutant Huntingtin and α-Synuclein* , 2007, Journal of Biological Chemistry.

[4]  Francesco Scaravilli,et al.  Inhibition of mTOR induces autophagy and reduces toxicity of polyglutamine expansions in fly and mouse models of Huntington disease , 2004, Nature Genetics.

[5]  M. Ashburner,et al.  The DrosDel Collection , 2004, Genetics.

[6]  Alexei Degterev,et al.  Regulation of Intracellular Accumulation of Mutant Huntingtin by Beclin 1* , 2006, Journal of Biological Chemistry.

[7]  Y. Moriyama,et al.  Bafilomycin A1 prevents maturation of autophagic vacuoles by inhibiting fusion between autophagosomes and lysosomes in rat hepatoma cell line, H-4-II-E cells. , 1998, Cell structure and function.

[8]  S. Schreiber,et al.  Small molecules enhance autophagy and reduce toxicity in Huntington's disease models. , 2007, Nature chemical biology.

[9]  N. Wood,et al.  Chronic lithium chloride treatment has variable effects on motor behaviour and survival of mice transgenic for the Huntington’s disease mutation , 2003, Brain Research Bulletin.

[10]  N. Mizushima,et al.  How to Interpret LC3 Immunoblotting , 2007, Autophagy.

[11]  Claire H. Michel,et al.  Lithium rescues toxicity of aggregate-prone proteins in Drosophila by perturbing Wnt pathway. , 2005, Human molecular genetics.

[12]  D. Rubinsztein,et al.  Rapamycin alleviates toxicity of different aggregate-prone proteins. , 2006, Human molecular genetics.

[13]  D. Rubinsztein,et al.  A molecular investigation of true dominance in Huntington’s disease , 1999, Journal of medical genetics.

[14]  Takeshi Noda,et al.  LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing , 2000, The EMBO journal.

[15]  N. Mizushima Methods for monitoring autophagy. , 2004, The international journal of biochemistry & cell biology.

[16]  D. Rubinsztein,et al.  Lithium induces autophagy by inhibiting inositol monophosphatase , 2005, The Journal of cell biology.

[17]  G. Mills,et al.  Glycogen Synthase Kinase 3β Is a Negative Regulator of Growth Factor-induced Activation of the c-Jun N-terminal Kinase* , 2004, Journal of Biological Chemistry.

[18]  C. Ross,et al.  What is the role of protein aggregation in neurodegeneration? , 2005, Nature Reviews Molecular Cell Biology.

[19]  D. Rubinsztein,et al.  Potential therapeutic applications of autophagy , 2007, Nature Reviews Drug Discovery.

[20]  Leslie Michels Thompson,et al.  Drosophila in the Study of Neurodegenerative Disease , 2006, Neuron.

[21]  C. Phiel,et al.  Molecular targets of lithium action. , 2003, Annual review of pharmacology and toxicology.

[22]  H. Manji,et al.  Lithium: A Molecular Transducer of Mood-Stabilization in the Treatment of Bipolar Disorder , 1998, Neuropsychopharmacology.

[23]  J. Hodges,et al.  Psychiatric symptoms and CAG repeats in neurologically asymptomatic Huntington's disease gene carriers , 2001, Psychiatry Research.

[24]  D. Rubinsztein,et al.  Glycogen Synthase Kinase-3β Inhibitors Prevent Cellular Polyglutamine Toxicity Caused by the Huntington's Disease Mutation* , 2002, The Journal of Biological Chemistry.

[25]  Ming You,et al.  TSC2 Integrates Wnt and Energy Signals via a Coordinated Phosphorylation by AMPK and GSK3 to Regulate Cell Growth , 2006, Cell.

[26]  Iris Salecker,et al.  Polyglutamine-Expanded Human Huntingtin Transgenes Induce Degeneration of Drosophila Photoreceptor Neurons , 1998, Neuron.

[27]  Takeshi Tokuhisa,et al.  Dissection of Autophagosome Formation Using Apg5-Deficient Mouse Embryonic Stem Cells , 2001, The Journal of cell biology.

[28]  D. Rubinsztein Lessons from animal models of Huntington's disease. , 2002, Trends in genetics : TIG.

[29]  Mauro Delorenzi,et al.  Mutant huntingtin's effects on striatal gene expression in mice recapitulate changes observed in human Huntington's disease brain and do not differ with mutant huntingtin length or wild-type huntingtin dosage. , 2007, Human molecular genetics.

[30]  Jeremy N. Skepper,et al.  α-Synuclein Is Degraded by Both Autophagy and the Proteasome* , 2003, Journal of Biological Chemistry.

[31]  T. P. Neufeld,et al.  Regulation of cellular growth by the Drosophila target of rapamycin dTOR. , 2000, Genes & development.