Faithful SGCE imprinting in iPSC-derived cortical neurons: an endogenous cellular model of myoclonus-dystonia.
In neuropathology research, induced pluripotent stem cell (iPSC)-derived neurons are considered a tool closely resembling the patient brain. Albeit in respect to epigenetics, this concept has been challenged. We generated iPSC-derived cortical neurons from myoclonus-dystonia patients with mutations (W100G and R102X) in the maternally imprinted ε-sarcoglycan (SGCE) gene and analysed properties such as imprinting, mRNA and protein expression. Comparison of the promoter during reprogramming and differentiation showed tissue-independent differential methylation. DNA sequencing with methylation-specific primers and cDNA analysis in patient neurons indicated selective expression of the mutated paternal SGCE allele. While fibroblasts only expressed the ubiquitous mRNA isoform, brain-specific SGCE mRNA and ε-sarcoglycan protein were detected in iPSC-derived control neurons. However, neuronal protein levels were reduced in both mutants. Our phenotypic characterization highlights the suitability of iPSC-derived cortical neurons with SGCE mutations for myoclonus-dystonia research and, in more general terms, prompts the use of iPSC-derived cellular models to study epigenetic mechanisms impacting on health and disease.
Institute of Neurogenetics, University of Lübeck, 23562 Lübeck, Germany.
Karen
K
Grütz
Institute of Neurogenetics, University of Lübeck, 23562 Lübeck, Germany.
Philip
P
Seibler
Institute of Neurogenetics, University of Lübeck, 23562 Lübeck, Germany.
Anne
A
Weissbach
Institute of Neurogenetics, University of Lübeck, 23562 Lübeck, Germany.
Katja
K
Lohmann
MRC Centre for Neuropsychiatric Genetics and Genomics, Institute of Psychological Medicine and Clinical Neurosciences, School of Medicine, Cardiff University, Cardiff CF24 4HQ, United Kingdom.
Francesca A
FA
Carlisle
MRC Centre for Neuropsychiatric Genetics and Genomics, Institute of Psychological Medicine and Clinical Neurosciences, School of Medicine, Cardiff University, Cardiff CF24 4HQ, United Kingdom.
Derek J
DJ
Blake
Institute of Neurogenetics, University of Lübeck, 23562 Lübeck, Germany.
Ana
A
Westenberger
Institute of Neurogenetics, University of Lübeck, 23562 Lübeck, Germany.
Christine
C
Klein
Institute of Neurogenetics, University of Lübeck, 23562 Lübeck, Germany.|Molecular and Functional Neurobiology Group, Luxembourg Centre for Systems Biomedicine, University of Luxembourg, L-4367 Belvaux, Luxembourg.
Anne
A
Grünewald
D002460:Cell Line; D019175:DNA Methylation; D020821:Dystonic Disorders / Q000235:genetics* / Q000378:metabolism; D005260:Female; D018392:Genomic Imprinting; D057809:HEK293 Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D008297:Male; D008954:Models, Biological; D009154:Mutation; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D049031:Sarcoglycans / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA
C526572:SGCE protein, human; D049031:Sarcoglycans