Autophagy Activation by Transcription Factor EB (TFEB) in Striatum of HDQ175/Q7 Mice.
Mutant huntingtin (mHTT) is encoded by the Huntington's disease (HD) gene and its accumulation in the brain contributes to HD pathogenesis. Reducing mHTT levels through activation of the autophagosome-lysosomal pathway may have therapeutic benefit. Transcription factor EB (TFEB) regulates lysosome biogenesis and autophagy.



To examine if increasing TFEB protein levels in HD mouse striatum induces autophagy and influences mHTT levels.



We introduced cDNA encoding TFEB with an HA tag (TFEB-HA) under the control of neuron specific synapsin 1 promoter into the striatum of 3 month old HDQ175/Q7 mice using adeno-associated virus AAV2/9. The levels of exogenous TFEB were analyzed using qPCR and Western blot. Proteins involved in autophagy, levels of huntingtin, and striatal-enriched proteins were examined using biochemical and/or immunohistochemical methods.



In HD mice expressing TFEB-HA, HA immunoreactivity distributed throughout the striatum in neuronal cell bodies and processes and preferentially in neuronal nuclei and overlapped with a loss of DARPP32 immunoreactivity. TFEB-HA mRNA and protein were detected in striatal lysates. There were increased levels of proteins involved with autophagosome/lysosome activity including LAMP-2A, LC3II, and cathepsin D and reduced levels of mutant HTT and the striatal enriched proteins DARPP32 and PDE10A. Compared to WT mice, HDQ175/Q7 mice had elevated levels of the ER stress protein GRP78/BiP and with TFEB-HA expression, increased levels of the astrocyte marker GFAP and pro-caspase 3.



These results suggest that TFEB expression in the striatum of HDQ175/Q7 mice stimulates autophagy and lysosome activity, and lowers mHTT, but may also increase a neuronal stress response.
Department of Neurology, Massachusetts General Hospital and Harvard Medical School, Charlestown, MA, USA.
Petr
P
Vodicka
Departments of Medicine and Cell Biology, University of Massachusetts, Worcester, MA, USA.
Kathryn
K
Chase
Department of Neurology, Massachusetts General Hospital and Harvard Medical School, Charlestown, MA, USA.
Maria
M
Iuliano
Department of Neurology, Massachusetts General Hospital and Harvard Medical School, Charlestown, MA, USA.
Adelaide
A
Tousley
Department of Neurology, Massachusetts General Hospital and Harvard Medical School, Charlestown, MA, USA.
Dana T
DT
Valentine
Department of Neurology, Massachusetts General Hospital and Harvard Medical School, Charlestown, MA, USA.
Ellen
E
Sapp
Department of Neurology, Massachusetts General Hospital and Harvard Medical School, Charlestown, MA, USA.
Kimberly B
KB
Kegel-Gleason
Department of Neurology, Horae Gene Therapy Center, University of Massachusetts Medical School, Albert Sherman Center, Worcester, MA, USA.
Miguel
M
Sena-Esteves
Departments of Medicine and Cell Biology, University of Massachusetts, Worcester, MA, USA.
Neil
N
Aronin
Department of Neurology, Massachusetts General Hospital and Harvard Medical School, Charlestown, MA, USA.
Marian
M
DiFiglia
D000818:Animals; D001343:Autophagy / Q000235:genetics*; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors / Q000235:genetics / Q000378:metabolism*; D002402:Cathepsin D / Q000378:metabolism; D002452:Cell Count; D003342:Corpus Striatum / Q000378:metabolism*; D004195:Disease Models, Animal; D051937:Dopamine and cAMP-Regulated Phosphoprotein 32 / Q000378:metabolism; D000091342:Endoplasmic Reticulum Chaperone BiP; D005786:Gene Expression Regulation / Q000235:genetics; D050883:HSC70 Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D000071058:Huntingtin Protein / Q000235:genetics; D006816:Huntington Disease / Q000235:genetics / Q000473:pathology*; D052119:Lysosomal-Associated Membrane Protein 2 / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000378:metabolism; D000090802:rab7 GTP-Binding Proteins
D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D051937:Dopamine and cAMP-Regulated Phosphoprotein 32; D000091342:Endoplasmic Reticulum Chaperone BiP; D050883:HSC70 Heat-Shock Proteins; C000717247:HSPA5 protein, human; C000717248:Hspa5 protein, mouse; D000071058:Huntingtin Protein; D052119:Lysosomal-Associated Membrane Protein 2; D009419:Nerve Tissue Proteins; C497929:Ppp1r1b protein, mouse; C064972:TFEB protein, human; D000090802:rab7 GTP-Binding Proteins; C119843:PDE10A protein, human; D010727:Phosphoric Diester Hydrolases; D002402:Cathepsin D; D020691:rab GTP-Binding Proteins
Adeno associated virus; GRP78/BiP; Huntington’s disease; LC3; TFEB; autophagy; huntingtin; neurodegeneration; striatum
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