title: Kinase Inhibitor Screening Identifies Cyclin-Dependent Kinases and Glycogen Synthase Kinase 3 as Potential Modulators of TDP-43 Cytosolic Accumulation during Cell Stress.
abstract: Abnormal processing of TAR DNA binding protein 43 (TDP-43) has been identified as a major factor in neuronal degeneration during amyotrophic lateral sclerosis (ALS) or frontotemporal lobar degeneration (FTLD). It is unclear how changes to TDP-43, including nuclear to cytosolic translocation and subsequent accumulation, are controlled in these diseases. TDP-43 is a member of the heterogeneous ribonucleoprotein (hnRNP) RNA binding protein family and is known to associate with cytosolic RNA stress granule proteins in ALS and FTLD. hnRNP trafficking and accumulation is controlled by the action of specific kinases including members of the mitogen-activated protein kinase (MAPK) pathway. However, little is known about how kinase pathways control TDP-43 movement and accumulation. In this study, we used an in vitro model of TDP-43-positve stress granule formation to screen for the effect of kinase inhibitors on TDP-43 accumulation. We found that while a number of kinase inhibitors, particularly of the MAPK pathways modulated both TDP-43 and the global stress granule marker, human antigen R (HuR), multiple inhibitors were more specific to TDP-43 accumulation, including inhibitors of cyclin-dependent kinases (CDKs) and glycogen synthase kinase 3 (GSK3). Close correlation was observed between effects of these inhibitors on TDP-43, hnRNP K and TIAR, but often with different effects on HuR accumulation. This may indicate a potential interaction between TDP-43, hnRNP K and TIAR. CDK inhibitors were also found to reverse pre-formed TDP-43-positive stress granules and both CDK and GSK3 inhibitors abrogated the accumulation of C-terminal TDP-43 (219-414) in transfected cells. Further studies are required to confirm the specific kinases involved and whether their action is through phosphorylation of the TDP-43 binding partner hnRNP K. This knowledge provides a valuable insight into the mechanisms controlling abnormal cytoplasmic TDP-43 accumulation and may herald new opportunities for kinase modulation-based therapeutic intervention in ALS and FTLD.
authors.affiliation: Department of Pathology, The University of Melbourne, Victoria, Australia and Florey Institute of Neuroscience and Mental Health, Parkville, Victoria, Australia.
authors.forename: Diane
authors.initials: D
authors.lastname: Moujalled
authors.forename: Janine L
authors.initials: JL
authors.lastname: James
authors.forename: Sarah J
authors.initials: SJ
authors.lastname: Parker
authors.forename: Grace E
authors.initials: GE
authors.lastname: Lidgerwood
authors.forename: Clare
authors.initials: C
authors.lastname: Duncan
authors.forename: Jodi
authors.initials: J
authors.lastname: Meyerowitz
authors.forename: Takashi
authors.initials: T
authors.lastname: Nonaka
authors.forename: Masato
authors.initials: M
authors.lastname: Hasegawa
authors.forename: Katja M
authors.initials: KM
authors.lastname: Kanninen
authors.forename: Alexandra
authors.initials: A
authors.lastname: Grubman
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Liddell
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Crouch
authors.forename: Anthony R
authors.initials: AR
authors.lastname: White
mesh_terms: D045744:Cell Line, Tumor; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D003600:Cytosol / Q000187:drug effects* / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D004353:Drug Evaluation, Preclinical; D005786:Gene Expression Regulation / Q000187:drug effects; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006801:Humans; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D021381:Protein Transport / Q000187:drug effects
chemical_list: D004268:DNA-Binding Proteins; D047428:Protein Kinase Inhibitors; C000598624:TARDBP protein, human; D018844:Cyclin-Dependent Kinases; D038362:Glycogen Synthase Kinase 3