@article{71656b86023c4ebbb61fa950f9fd2794,
title = "Individual epigenetic status of the pathogenic D4Z4 macrosatellite correlates with disease in facioscapulohumeral muscular dystrophy",
abstract = "Background: Both forms of facioscapulohumeral muscular dystrophy (FSHD) are associated with aberrant epigenetic regulation of the chromosome 4q35 D4Z4 macrosatellite. Chromatin changes due to large deletions of heterochromatin (FSHD1) or mutations in chromatin regulatory proteins (FSHD2) lead to relaxation of epigenetic repression and increased expression of the deleterious double homeobox 4 (DUX4) gene encoded within the distal D4Z4 repeat. However, many individuals with the genetic requirements for FSHD remain asymptomatic throughout their lives. Here we investigated family cohorts of FSHD1 individuals who were either affected (manifesting) or without any discernible weakness (nonmanifesting/asymptomatic) and their unaffected family members to determine if individual epigenetic status and stability of repression at the contracted 4q35 D4Z4 array in myocytes correlates with FSHD disease. Results: Family cohorts were analyzed for DNA methylation on the distal pathogenic 4q35 D4Z4 repeat on permissive A-type subtelomeres. We found DNA hypomethylation in FSHD1-affected subjects, hypermethylation in healthy controls, and distinctly intermediate levels of methylation in nonmanifesting subjects. We next tested if these differences in DNA methylation had functional relevance by assaying DUX4-fl expression and the stability of epigenetic repression of DUX4-fl in myogenic cells. Treatment with drugs that alter epigenetic status revealed that healthy cells were refractory to treatment, maintaining stable repression of DUX4, while FSHD1-affected cells were highly responsive to treatment and thus epigenetically poised to express DUX4. Myocytes from nonmanifesting subjects had significantly higher levels of DNA methylation and were more resistant to DUX4 activation in response to epigenetic drug treatment than cells from FSHD1-affected first-degree relatives containing the same contraction, indicating that the epigenetic status of the contracted D4Z4 array is reflective of disease. Conclusions: The epigenetic status of the distal 4qA D4Z4 repeat correlates with FSHD disease; FSHD-affected subjects have hypomethylation, healthy unaffected subjects have hypermethylation, and nonmanifesting subjects have characteristically intermediate methylation. Thus, analysis of DNA methylation at the distal D4Z4 repeat could be used as a diagnostic indicator of developing clinical FSHD. In addition, the stability of epigenetic repression upstream of DUX4 expression is a key regulator of disease and a viable therapeutic target.",
keywords = "D4Z4, DNA methylation, DUX4, Decitabine, Disease modifier, Epiallele, Epigenetic modifier, FSHD, Muscular dystrophy",
author = "Jones, {Takako I.} and King, {Oliver D.} and Himeda, {Charis L.} and Sachiko Homma and Chen, {Jennifer C.J.} and Beermann, {Mary Lou} and Chi Yan and Emerson, {Charles P.} and Miller, {Jeffrey B.} and Wagner, {Kathryn R.} and Jones, {Peter L.}",
note = "Funding Information: We thank members of the Sen. Paul D. Wellstone Muscular Dystrophy Cooperative Research Center for FSHD Research for deriving the original cultures of myogenic cells [33,45,46]: Dr. Genila Bibat (Kennedy-Krieger Institute and Johns Hopkins School of Medicine) for coordinating the clinical aspects of the study and Ms. Kendal Hanger (University of Massachusetts Medical School) for the preparation of myogenic cells from biopsies. The authors thank the participating subjects and their families, and Mr. Daniel P. Perez and the FSH Society for subject recruitment and funding of travel costs for subject participation, and are grateful to Dr. Stephen Tapscott and Dr. Linda N. Geng (Fred Hutchinson Cancer Research Center) for generously providing initial samples of DUX4-FL mAbs. Genotyping was performed by Steve Moore at the University of Iowa. This work was funded by grant R01AR062587 from the National Institute of Arthritis, Musculoskeletal and Skin Diseases (NIAMS) NIH awarded to PLJ, NIAMS(National Institute of Arthritis, Musculoskeletal and Skin Diseases) grant R01AR060328 awarded to JBM, grant AFM15700 from the Association Fran{\c c}aise contra les myopathies awarded to PLJ and JBM, grant MDA216422 from the Muscular Dystrophy Association (MDA) awarded to JBM, grant #MDA216652 awarded to CPE, and the Senator Paul D. Wellstone Muscular Dystrophy Cooperative Research Center for FSHD Research grant U54HD060848 from the Eunice Kennedy Shriver National Institute for Child Health and Human Development. The authors thank the Chris Carrino Foundation for FSHD and the Thoracic Foundation (Boston, MA, USA) for their support. The content is solely the responsibility of the authors and does not necessarily represent the official views of the funding agencies. Funding Information: We thank members of the Sen. Paul D. Wellstone Muscular Dystrophy Cooperative Research Center for FSHD Research for deriving the original cultures of myogenic cells [33,45,46]: Dr. Genila Bibat (Kennedy-Krieger Institute and Johns Hopkins School of Medicine) for coordinating the clinical aspects of the study and Ms. Kendal Hanger (University of Massachusetts Medical School) for the preparation of myogenic cells from biopsies. The authors thank the participating subjects and their families, and Mr. Daniel P. Perez and the FSH Society for subject recruitment and funding of travel costs for subject participation, and are grateful to Dr. Stephen Tapscott and Dr. Linda N. Geng (Fred Hutchinson Cancer Research Center) for generously providing initial samples of DUX4-FL mAbs. Genotyping was performed by Steve Moore at the University of Iowa. This work was funded by grant R01AR062587 from the National Institute of Arthritis, Musculoskeletal and Skin Diseases (NIAMS) NIH awarded to PLJ, NIAMS grant R01AR060328 awarded to JBM, grant AFM15700 from the Association Fran{\c c}aise contra les myopathies awarded to PLJ and JBM, grant MDA216422 from the Muscular Dystrophy Association (MDA) awarded to JBM, grant #MDA216652 awarded to CPE, and the Senator Paul D. Wellstone Muscular Dystrophy Cooperative Research Center for FSHD Research grant U54HD060848 from the Eunice Kennedy Shriver National Institute for Child Health and Human Development. The authors thank the Chris Carrino Foundation for FSHD and the Thoracic Foundation (Boston, MA, USA) for their support. The content is solely the responsibility of the authors and does not necessarily represent the official views of the funding agencies. Publisher Copyright: {\textcopyright} 2015 Jones et al.",
year = "2015",
doi = "10.1186/s13148-015-0072-6",
language = "English (US)",
volume = "7",
journal = "Clinical Epigenetics",
issn = "1868-7075",
publisher = "Springer Verlag",
number = "1",
}