Aberrant expression of miR-218 and miR-204 in human mesial temporal lobe epilepsy and hippocampal sclerosis—Convergence on axonal guidance
Sanne S. Kaalund
Department of Cellular and Molecular Medicine, Wilhelm Johannsen Center for Functional Genome Research, University of Copenhagen, Copenhagen, Denmark
Research Laboratory for Stereology and Neuroscience, Bispebjerg Hospital, Copenhagen, Denmark
Search for more papers by this authorMorten T. Venø
Department of Molecular Biology and Genetics, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Aarhus, Denmark
Search for more papers by this authorMads Bak
Department of Cellular and Molecular Medicine, Wilhelm Johannsen Center for Functional Genome Research, University of Copenhagen, Copenhagen, Denmark
Search for more papers by this authorRikke S. Møller
Department of Cellular and Molecular Medicine, Wilhelm Johannsen Center for Functional Genome Research, University of Copenhagen, Copenhagen, Denmark
Danish Epilepsy Center, Dianalund, Denmark
Institute for Regional Health Services, University of Southern Denmark, Odense, Denmark
Search for more papers by this authorHenning Laursen
Laboratory of Neuropathology, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark
Search for more papers by this authorFlemming Madsen
Department of Neurosurgery, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark
Search for more papers by this authorHelle Broholm
Laboratory of Neuropathology, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark
Search for more papers by this authorBjørn Quistorff
Department of Biomedical Sciences, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark
Search for more papers by this authorPeter Uldall
Department of Pediatrics, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark
Search for more papers by this authorNiels Tommerup
Department of Cellular and Molecular Medicine, Wilhelm Johannsen Center for Functional Genome Research, University of Copenhagen, Copenhagen, Denmark
Search for more papers by this authorSakari Kauppinen
Department of Haematology, Aalborg University Hospital, Aalborg, Denmark
Search for more papers by this authorAnne Sabers
Department of Neurology, The Epilepsy Clinic, Copenhagen University Hospital, Rigshospitalet, Copenhagen, Denmark
Search for more papers by this authorKees Fluiter
Department of Neurogenetics, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands
Search for more papers by this authorLisbeth B. Møller
Department of Clinical Genetics, Applied Human Molecular Genetics, Kennedy Center, Copenhagen University Hospital, Rigshospitalet, Glostrup, Denmark
Search for more papers by this authorAnne Y. Nossent
Laboratory for Molecular Cardiology, Department of Biomedical Sciences, The Danish National Research Foundation Centre for Cardiac Arrhythmia, University of Copenhagen, Copenhagen, Denmark
Search for more papers by this authorAsli Silahtaroglu
Department of Cellular and Molecular Medicine, Wilhelm Johannsen Center for Functional Genome Research, University of Copenhagen, Copenhagen, Denmark
Search for more papers by this authorJørgen Kjems
Department of Molecular Biology and Genetics, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Aarhus, Denmark
Search for more papers by this authorEleonora Aronica
SEIN–Stichting Epilepsie Instellingen Nederland, Heemstede, The Netherlands
Department of (Neuro) Pathology, AZ Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands
The senior authors who contributed equally to the manuscript.Search for more papers by this authorCorresponding Author
Zeynep Tümer
Department of Cellular and Molecular Medicine, Wilhelm Johannsen Center for Functional Genome Research, University of Copenhagen, Copenhagen, Denmark
Department of Clinical Genetics, Applied Human Molecular Genetics, Kennedy Center, Copenhagen University Hospital, Rigshospitalet, Glostrup, Denmark
The senior authors who contributed equally to the manuscript.Address correspondence to Zeynep Tümer, Center for Applied Human Molecular Genetics, The Kennedy Center, Gl. Landevej 7, 2600 Glostrup, Denmark. E-mail: [email protected]Search for more papers by this authorSanne S. Kaalund
Department of Cellular and Molecular Medicine, Wilhelm Johannsen Center for Functional Genome Research, University of Copenhagen, Copenhagen, Denmark
Research Laboratory for Stereology and Neuroscience, Bispebjerg Hospital, Copenhagen, Denmark
Search for more papers by this authorMorten T. Venø
Department of Molecular Biology and Genetics, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Aarhus, Denmark
Search for more papers by this authorMads Bak
Department of Cellular and Molecular Medicine, Wilhelm Johannsen Center for Functional Genome Research, University of Copenhagen, Copenhagen, Denmark
Search for more papers by this authorRikke S. Møller
Department of Cellular and Molecular Medicine, Wilhelm Johannsen Center for Functional Genome Research, University of Copenhagen, Copenhagen, Denmark
Danish Epilepsy Center, Dianalund, Denmark
Institute for Regional Health Services, University of Southern Denmark, Odense, Denmark
Search for more papers by this authorHenning Laursen
Laboratory of Neuropathology, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark
Search for more papers by this authorFlemming Madsen
Department of Neurosurgery, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark
Search for more papers by this authorHelle Broholm
Laboratory of Neuropathology, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark
Search for more papers by this authorBjørn Quistorff
Department of Biomedical Sciences, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark
Search for more papers by this authorPeter Uldall
Department of Pediatrics, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark
Search for more papers by this authorNiels Tommerup
Department of Cellular and Molecular Medicine, Wilhelm Johannsen Center for Functional Genome Research, University of Copenhagen, Copenhagen, Denmark
Search for more papers by this authorSakari Kauppinen
Department of Haematology, Aalborg University Hospital, Aalborg, Denmark
Search for more papers by this authorAnne Sabers
Department of Neurology, The Epilepsy Clinic, Copenhagen University Hospital, Rigshospitalet, Copenhagen, Denmark
Search for more papers by this authorKees Fluiter
Department of Neurogenetics, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands
Search for more papers by this authorLisbeth B. Møller
Department of Clinical Genetics, Applied Human Molecular Genetics, Kennedy Center, Copenhagen University Hospital, Rigshospitalet, Glostrup, Denmark
Search for more papers by this authorAnne Y. Nossent
Laboratory for Molecular Cardiology, Department of Biomedical Sciences, The Danish National Research Foundation Centre for Cardiac Arrhythmia, University of Copenhagen, Copenhagen, Denmark
Search for more papers by this authorAsli Silahtaroglu
Department of Cellular and Molecular Medicine, Wilhelm Johannsen Center for Functional Genome Research, University of Copenhagen, Copenhagen, Denmark
Search for more papers by this authorJørgen Kjems
Department of Molecular Biology and Genetics, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Aarhus, Denmark
Search for more papers by this authorEleonora Aronica
SEIN–Stichting Epilepsie Instellingen Nederland, Heemstede, The Netherlands
Department of (Neuro) Pathology, AZ Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands
The senior authors who contributed equally to the manuscript.Search for more papers by this authorCorresponding Author
Zeynep Tümer
Department of Cellular and Molecular Medicine, Wilhelm Johannsen Center for Functional Genome Research, University of Copenhagen, Copenhagen, Denmark
Department of Clinical Genetics, Applied Human Molecular Genetics, Kennedy Center, Copenhagen University Hospital, Rigshospitalet, Glostrup, Denmark
The senior authors who contributed equally to the manuscript.Address correspondence to Zeynep Tümer, Center for Applied Human Molecular Genetics, The Kennedy Center, Gl. Landevej 7, 2600 Glostrup, Denmark. E-mail: [email protected]Search for more papers by this authorSummary
Objective
Mesial temporal lobe epilepsy (MTLE) is one of the most common types of the intractable epilepsies and is most often associated with hippocampal sclerosis (HS), which is characterized by pronounced loss of hippocampal pyramidal neurons. microRNAs (miRNAs) have been shown to be dysregulated in epilepsy and neurodegenerative diseases, and we hypothesized that miRNAs could be involved in the pathogenesis of MTLE and HS.
Methods
miRNA expression was quantified in hippocampal specimens from human patients using miRNA microarray and quantitative real-time polymerase chain reaction RT-PCR, and by RNA-seq on fetal brain specimens from domestic pigs. In situ hybridization was used to show the spatial distribution of miRNAs in the human hippocampus. The potential effect of miRNAs on targets genes was investigated using the dual luciferase reporter gene assay.
Results
miRNA expression profiling showed that 25 miRNAs were up-regulated and 5 were down-regulated in hippocampus biopsies of MTLE/HS patients compared to controls. We showed that miR-204 and miR-218 were significantly down-regulated in MTLE and HS, and both were expressed in neurons in all subfields of normal hippocampus. Moreover, miR-204 and miR-218 showed strong changes in expression during fetal development of the hippocampus in pigs, and we identified four target genes, involved in axonal guidance and synaptic plasticity, ROBO1, GRM1, SLC1A2, and GNAI2, as bona fide targets of miR-218. GRM1 was also shown to be a direct target of miR-204.
Significance
miR-204 and miR-218 are developmentally regulated in the hippocampus and may contribute to the molecular mechanisms underlying the pathogenesis of MTLE and HS.
Supporting Information
| Filename | Description |
|---|---|
| epi12839-sup-0001-TableS1.docxWord document, 16.1 KB | Table S1. Luciferase primer sequences for PCR amplification of 3′UTRs and pre-miRNA. |
| epi12839-sup-0002-TableS2.docxWord document, 15.3 KB | Table S2. Selected miRNA target genes. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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