Christin Krause1,2, Susen Schaake3, Karen Grütz3, Helen Sievert1, Charles Jourdan Reyes3, Inke R König4, Björn-Hergen Laabs4, Roland Dominic Jamora5, Raymond L Rosales6, Cid Czarina E Diesta7, Jelena Pozojevic2,8, Timo Gemoll9, Ana Westenberger3, Frank J Kaiser8,10, Christine Klein3, Henriette Kirchner1,2. 1. Institute for Human Genetics, Division Epigenetics & Metabolism, University of Lübeck, Lübeck, Germany. 2. Center of Brain, Behavior and Metabolism (CBBM), University of Lübeck, Ratzeburger Allee 160, 23562, Lübeck, Germany. 3. Institute of Neurogenetics, University of Lübeck, Lübeck, Germany. 4. Institute of Medical Biometry and Statistics, University of Lübeck, Lübeck, Germany. 5. Department of Neurosciences, College of Medicine - Philippine General Hospital, University of the Philippines, Manila, Philippines. 6. University of Santo Tomas Hospital, Manila, Philippines. 7. Department of Neurosciences, Movement Disorders Clinic, Makati Medical Center, Makati City, Philippines. 8. Section for Functional Genetics, Institute for Human Genetics, University of Lübeck, Lübeck, Germany. 9. Section for Translational Surgical Oncology and Biobanking, Department of Surgery, University of Lübeck and University Hospital Schleswig-Holstein, Kiel, Germany. 10. Institute of Human Genetics, University Hospital Essen, University Duisburg-Essen, Essen, Germany.
Abstract
BACKGROUND: X-linked dystonia-parkinsonism is a neurodegenerative movement disorder. The underlying molecular basis has still not been completely elucidated, but likely involves dysregulation of TAF1 expression. In X-linked dystonia-parkinsonism, 3 disease-specific single-nucleotide changes (DSCs) introduce (DSC12) or abolish (DSC2 and DSC3) CpG dinucleotides and consequently sites of putative DNA methylation. Because transcriptional regulation tightly correlates with specific epigenetic marks, we investigated the role of DNA methylation in the pathogenesis of X-linked dystonia-parkinsonism. METHODS: DNA methylation at DSC12, DSC3, and DSC2 was quantified by bisulfite pyrosequencing in DNA from peripheral blood leukocytes, fibroblasts, induced pluripotent stem cell-derived cortical neurons and brain tissue from X-linked dystonia-parkinsonism patients and age- and sex-matched healthy Filipino controls in a prospective study. RESULTS: Compared with controls, X-linked dystonia-parkinsonism patients showed striking differences in DNA methylation at the 3 investigated CpG sites. Using methylation-sensitive luciferase reporter gene assays and immunoprecipitation, we demonstrated (1) that lack of DNA methylation because of DSC2 and DSC3 affects gene promoter activity and (2) that methylation at all 3 investigated CpG sites alters DNA-protein interaction. Interestingly, DSC3 decreased promoter activity per se compared with wild type, and promoter activity further decreased when methylation was present. Moreover, we identified specific binding of proteins to the investigated DSCs that are associated with splicing and RNA and DNA binding. CONCLUSIONS: We identified altered DNA methylation in X-linked dystonia-parkinsonism patients as a possible additional mechanism modulating TAF1 expression and putative novel targets for future therapies using DNA methylation-modifying agents.
BACKGROUND:X-linked dystonia-parkinsonism is a neurodegenerative movement disorder. The underlying molecular basis has still not been completely elucidated, but likely involves dysregulation of TAF1 expression. In X-linked dystonia-parkinsonism, 3 disease-specific single-nucleotide changes (DSCs) introduce (DSC12) or abolish (DSC2 and DSC3) CpG dinucleotides and consequently sites of putative DNA methylation. Because transcriptional regulation tightly correlates with specific epigenetic marks, we investigated the role of DNA methylation in the pathogenesis of X-linked dystonia-parkinsonism. METHODS: DNA methylation at DSC12, DSC3, and DSC2 was quantified by bisulfite pyrosequencing in DNA from peripheral blood leukocytes, fibroblasts, induced pluripotent stem cell-derived cortical neurons and brain tissue from X-linked dystonia-parkinsonismpatients and age- and sex-matched healthy Filipino controls in a prospective study. RESULTS: Compared with controls, X-linked dystonia-parkinsonismpatients showed striking differences in DNA methylation at the 3 investigated CpG sites. Using methylation-sensitive luciferase reporter gene assays and immunoprecipitation, we demonstrated (1) that lack of DNA methylation because of DSC2 and DSC3 affects gene promoter activity and (2) that methylation at all 3 investigated CpG sites alters DNA-protein interaction. Interestingly, DSC3 decreased promoter activity per se compared with wild type, and promoter activity further decreased when methylation was present. Moreover, we identified specific binding of proteins to the investigated DSCs that are associated with splicing and RNA and DNA binding. CONCLUSIONS: We identified altered DNA methylation in X-linked dystonia-parkinsonismpatients as a possible additional mechanism modulating TAF1 expression and putative novel targets for future therapies using DNA methylation-modifying agents.
Authors: Jelena Pozojevic; Shela Marie Algodon; Joseph Neos Cruz; Joanne Trinh; Norbert Brüggemann; Joshua Laß; Karen Grütz; Susen Schaake; Ronnie Tse; Veronica Yumiceba; Nathalie Kruse; Kristin Schulz; Varun K A Sreenivasan; Raymond L Rosales; Roland Dominic G Jamora; Cid Czarina E Diesta; Jakob Matschke; Markus Glatzel; Philip Seibler; Kristian Händler; Aleksandar Rakovic; Henriette Kirchner; Malte Spielmann; Frank J Kaiser; Christine Klein; Ana Westenberger Journal: Int J Mol Sci Date: 2022-02-17 Impact factor: 5.923