J Stroke.
2018 Sep;20(3):350-361. 10.5853/jos.2018.00962.
Aberrant Promoter Hypomethylation of Sortilin 1: A Moyamoya Disease Biomarker
- Affiliations
-
- 1Department of Biochemistry, Ewha Womans University College of Medicine, Seoul, Korea. nsthomas@snu.ac.kr ahnj@ewha.ac.kr
- 2Department of Anatomy, Seoul National University College of Medicine, Seoul, Korea.
- 3Division of Pediatric Neurosurgery, Seoul National University Children's Hospital, Seoul National University College of Medicine, Seoul, Korea. nsthomas@snu.ac.kr ahnj@ewha.ac.kr
- 4Suncheon National University College of Pharmacy, Suncheon, Korea.
- 5Department of Molecular Medicine, Ewha Womans University College of Medicine, Seoul, Korea.
- 6Department of Pharmacology, Ewha Womans University College of Medicine, Seoul, Korea.
- KMID: 2428770
- DOI: http://doi.org/10.5853/jos.2018.00962
Abstract
- BACKGROUND AND PURPOSE
The pathogenesis of moyamoya disease (MMD) remains poorly understood, and no reliable molecular biomarkers for MMD have been identified to date. The present study aimed to identify epigenetic biomarkers for use in the diagnosis of MMD.
METHODS
We performed integrated analyses of gene expression profiles and DNA methylation profiles in endothelial colony forming cells (ECFCs) from three patients with MMD and two healthy individuals. Candidate gene mRNA expression and DNA methylation status were further validated using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and pyrosequencing analysis of an expanded ECFC sample set from nine patients with MMD and ten controls. We evaluated the diagnostic accuracy of the potential biomarkers identified here using receiver operating characteristic curve analyses and further measured major angiogenic factor expression levels using a tube formation assay and RT-qPCR.
RESULTS
Five candidate genes were selected via integrated analysis; all five were upregulated by hypomethylation of specific promoter CpG sites. After further validation in an expanded sample set, we identified a candidate biomarker gene, sortilin 1 (SORT1). DNA methylation status at a specific SORT1 promoter CpG site in ECFCs readily distinguished patients with MMD from the normal controls with high accuracy (area under the curve 0.98, sensitivity 83.33%, specificity 100%). Furthermore, SORT1 overexpression suppressed endothelial cell tube formation and modulated major angiogenic factor and matrix metalloproteinase-9 expression, implying SORT1 involvement in MMD pathogenesis.
CONCLUSIONS
Our findings suggest that DNA methylation status at the SORT1 promoter CpG site may be a potential biomarker for MMD.
Keyword
MeSH Terms
-
Angiogenesis Inducing Agents
Biomarkers
Diagnosis
DNA Methylation
Endothelial Cells
Epigenomics
Humans
Matrix Metalloproteinase 9
Moyamoya Disease*
Polymerase Chain Reaction
RNA, Messenger
ROC Curve
Sensitivity and Specificity
Transcriptome
Angiogenesis Inducing Agents
Biomarkers
Matrix Metalloproteinase 9
RNA, Messenger
