title: SOX11 identified by target gene evaluation of miRNAs differentially expressed in focal and non-focal brain tissue of therapy-resistant epilepsy patients.
abstract: MicroRNAs (miRNAs) are small non-coding RNAs that post-transcriptionally control the expression of their target genes via RNA interference. There is increasing evidence that expression of miRNAs is dysregulated in neuronal disorders, including epilepsy, a chronic neurological disorder characterized by spontaneous recurrent seizures. Mesial temporal lobe epilepsy (MTLE) is a common type of focal epilepsy in which disease-induced abnormalities of hippocampal neurogenesis in the subgranular zone as well as gliosis and neuronal cell loss in the cornu ammonis area are reported. We hypothesized that in MTLE altered miRNA-mediated regulation of target genes could be involved in hippocampal cell remodeling. A miRNA screen was performed in hippocampal focal and non-focal brain tissue samples obtained from the temporal neocortex (both n=8) of MTLE patients. Out of 215 detected miRNAs, two were differentially expressed (hsa-miR-34c-5p: mean increase of 5.7 fold (p=0.014), hsa-miR-212-3p: mean decrease of 76.9% (p=0.0014)). After in-silico target gene analysis and filtering, reporter gene assays confirmed RNA interference for hsa-miR-34c-5p with 3'-UTR sequences of GABRA3, GRM7 and GABBR2 and for hsa-miR-212-3p with 3'-UTR sequences of SOX11, MECP2, ADCY1 and ABCG2. Reporter gene assays with mutated 3'-UTR sequences of the transcription factor SOX11 identified two different binding sites for hsa-miR-212-3p and its primary transcript partner hsa-miR-132-3p. Additionally, there was an inverse time-dependent expression of Sox11 and miR-212-3p as well as miR-132-3p in rat neonatal cortical neurons. Transfection of neurons with anti-miRs for miR-212-3p and miR-132-3p suggest that both miRNAs work synergistically to control Sox11 expression. Taken together, these results suggest that differential miRNA expression in neurons could contribute to an altered function of the transcription factor SOX11 and other genes in the setting of epilepsy, resulting not only in impaired neural differentiation, but also in imbalanced neuronal excitability and accelerated drug export.
authors.affiliation: School of Pharmacy, Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, CA, USA; Institute of Experimental and Clinical Pharmacology, University Hospital Schleswig-Holstein, Campus Kiel, Germany. Electronic address: haenisch@pharmakologie.uni-kiel.de.
authors.forename: Sierk
authors.initials: S
authors.lastname: Haenisch
authors.affiliation: Department of Nuclear Medicine, Molecular Imaging and Therapy, University Hospital Schleswig-Holstein, Campus Kiel, Germany.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: School of Pharmacy, Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, CA, USA.
authors.forename: Aparna
authors.initials: A
authors.lastname: Chhibber
authors.affiliation: Swedish Epilepsy Center, Neuroscience Institute, Seattle, WA, USA.
authors.forename: Kitti
authors.initials: K
authors.lastname: Kaiboriboon
authors.affiliation: Clinical Pharmacy Services, Medical Center, University of California, San Francisco, CA, USA; School of Pharmacy, Department of Clinical Pharmacy, University of California, San Francisco, CA, USA.
authors.forename: Lynn V
authors.initials: LV
authors.lastname: Do
authors.affiliation: Department of Neuropathology, Institute of Pathology, University of Greifswald, Greifswald, Germany.
authors.forename: Silke
authors.initials: S
authors.lastname: Vogelgesang
authors.affiliation: Department of Neurological Surgery and Neurology, School of Medicine, Indiana University, Indianapolis, IN, USA.
authors.forename: Nicholas M
authors.initials: NM
authors.lastname: Barbaro
authors.affiliation: School of Pharmacy, Department of Clinical Pharmacy, University of California, San Francisco, CA, USA; School of Medicine, Department of Neurology, University of California, San Francisco, CA, USA.
authors.forename: Brian K
authors.initials: BK
authors.lastname: Alldredge
authors.affiliation: School of Medicine, Department of Neurology, University of California, San Francisco, CA, USA.
authors.forename: Daniel H
authors.initials: DH
authors.lastname: Lowenstein
authors.affiliation: Institute of Experimental and Clinical Pharmacology, University Hospital Schleswig-Holstein, Campus Kiel, Germany.
authors.forename: Ingolf
authors.initials: I
authors.lastname: Cascorbi
authors.affiliation: School of Pharmacy, Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, CA, USA.
authors.forename: Deanna L
authors.initials: DL
authors.lastname: Kroetz
mesh_terms: D000328:Adult; D000818:Animals; D001921:Brain / Q000378:metabolism*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D003198:Computer Simulation; D000069279:Drug Resistant Epilepsy / Q000235:genetics* / Q000473:pathology*; D005260:Female; D020869:Gene Expression Profiling; D018197:Hepatoblastoma / Q000473:pathology; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000378:metabolism*; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D025341:Principal Component Analysis; D051381:Rats; D055751:SOXC Transcription Factors / Q000235:genetics* / Q000378:metabolism*; D013997:Time Factors; D055815:Young Adult
chemical_list: D035683:MicroRNAs; C097473:SOX11 protein, human; D055751:SOXC Transcription Factors
keywords: Epilepsy; GABA; Glutamate; MicroRNA; Neurogenesis; Neuronal differentiation; SOX11; Temporal lobe; hsa-miR-34c-5p; miR-132-3p; miR-212-3p