title: Epilepsy-causing sequence variations in SIK1 disrupt synaptic activity response gene expression and affect neuronal morphology.
abstract: SIK1 syndrome is a newly described developmental epilepsy disorder caused by heterozygous mutations in the salt-inducible kinase SIK1. To better understand the pathophysiology of SIK1 syndrome, we studied the effects of SIK1 pathogenic sequence variations in human neurons. Primary human fetal cortical neurons were transfected with a lentiviral vector to overexpress wild-type and mutant SIK1 protein. We evaluated the transcriptional activity of known downstream gene targets in neurons expressing mutant SIK1 compared with wild type. We then assayed neuronal morphology by measuring neurite length, number and branching. Truncating SIK1 sequence variations were associated with abnormal MEF2C transcriptional activity and decreased MEF2C protein levels. Epilepsy-causing SIK1 sequence variations were associated with significantly decreased expression of ARC (activity-regulated cytoskeletal-associated) and other synaptic activity response element genes. Assay of mRNA levels for other MEF2C target genes NR4A1 (Nur77) and NRG1, found significantly, decreased the expression of these genes as well. The missense p.(Pro287Thr) SIK1 sequence variation was associated with abnormal neuronal morphology, with significant decreases in mean neurite length, mean number of neurites and a significant increase in proximal branches compared with wild type. Epilepsy-causing SIK1 sequence variations resulted in abnormalities in the MEF2C-ARC pathway of neuronal development and synapse activity response. This work provides the first insights into the mechanisms of pathogenesis in SIK1 syndrome, and extends the ARX-MEF2C pathway in the pathogenesis of developmental epilepsy.
authors.affiliation: Department of Biomedical Genetics, University of Rochester Medical Center, Rochester, NY, USA.
authors.forename: Christoph
authors.initials: C
authors.lastname: Pröschel
authors.affiliation: Department of Pediatrics, University of Rochester Medical Center, Rochester, NY, USA.|Center for Neural Development and Disease, University of Rochester Medical Center, Rochester, NY, USA.
authors.forename: Jeanne N
authors.initials: JN
authors.lastname: Hansen
authors.affiliation: Center for Neural Development and Disease, University of Rochester Medical Center, Rochester, NY, USA.
authors.forename: Adil
authors.initials: A
authors.lastname: Ali
authors.affiliation: Center for Neural Development and Disease, University of Rochester Medical Center, Rochester, NY, USA.
authors.forename: Emily
authors.initials: E
authors.lastname: Tuttle
authors.affiliation: Department of Biomedical Genetics, University of Rochester Medical Center, Rochester, NY, USA.
authors.forename: Michelle
authors.initials: M
authors.lastname: Lacagnina
authors.affiliation: Department of Biomedical Genetics, University of Rochester Medical Center, Rochester, NY, USA.
authors.forename: Georgia
authors.initials: G
authors.lastname: Buscaglia
authors.affiliation: Center for Neural Development and Disease, University of Rochester Medical Center, Rochester, NY, USA.|Departments of Neurology and Neuroscience, University of Rochester Medical Center, Rochester, NY, USA.
authors.forename: Marc W
authors.initials: MW
authors.lastname: Halterman
authors.affiliation: Department of Biomedical Genetics, University of Rochester Medical Center, Rochester, NY, USA.|Department of Pediatrics, University of Rochester Medical Center, Rochester, NY, USA.|Center for Neural Development and Disease, University of Rochester Medical Center, Rochester, NY, USA.|Departments of Neurology and Neuroscience, University of Rochester Medical Center, Rochester, NY, USA.
authors.forename: Alex R
authors.initials: AR
authors.lastname: Paciorkowski
mesh_terms: D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism; D004827:Epilepsy / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D057809:HEK293 Cells; D006801:Humans; D064326:MEF2 Transcription Factors / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D020890:Neuregulin-1 / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000378:metabolism* / Q000473:pathology / Q000502:physiology; D057105:Nuclear Receptor Subfamily 4, Group A, Member 1 / Q000235:genetics / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism; D009435:Synaptic Transmission
chemical_list: D003598:Cytoskeletal Proteins; D064326:MEF2 Transcription Factors; C578040:MEF2C protein, human; C539821:NR4A1 protein, human; C094131:NRG1 protein, human; D009419:Nerve Tissue Proteins; D020890:Neuregulin-1; D057105:Nuclear Receptor Subfamily 4, Group A, Member 1; C092069:activity regulated cytoskeletal-associated protein; D017346:Protein Serine-Threonine Kinases; C509080:SIK1 protein, human