Facioscapulohumeral muscular dystrophy (FSHD) is a unique disorder involving shortening of an array of tandem 3.3-kb repeats. Unaffected individuals have 11-100 copies of this repeat, D4Z4, at both allelic subtelomeric regions on the long arm of chromosome 4 (at 4q35). Patients afflicted with this progressive, debilitating and painful disease have only 1-10 copies of the repeat on one of their chromosome 4 homologues. Almost identical arrays of D4Z4 repeats embedded in extremely similar sequences on both sides of the array for 25-45 kb are located at the subtelomeric end of the long arm of chromosome 10 but although these also can be present in 1-100 copies, there is no phenotype associated with short D4Z4 arrays on chromosome 10. Much evidence suggests that a short D4Z4 array on chromosome 4 causes FSHD by abnormally altering expression of a rather distant gene at 4q35. This research involves analyzing the nature of the chromatin and chromatin proteins in the D4Z4 arrays and in 4q35 gene regions and looking for long-distance looping interactions between the array and promoter regions of candidate FSHD genes as well as between the ends of the array. The cells to be analyzed will be diploid myoblasts, myotubes induced from myoblasts, and heterologous cell types, namely, lymphoblastoid cell lines and diploid fibroblasts. The cultures will be derived from FSHD patient samples, which will continue to be collected during this study, as well as from disease-controls; the known sizes of their D4Z4 arrays will be compared to the properties of chromatin at 4q35. In vivo DNasel and dimethyl sulfate footprinting, electrophoretic mobility shift assays, chromatin immunoprecipitation assays, immunocytochemistry, and two new assays developed to monitor long-range chromatin interactions will be the main techniques used in this study. The proposed research should elucidate new aspects of long-distance control of gene expression as well as lending clinically useful insights into this currently intractable disease. ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS048859-03
Application #
7052794
Study Section
Skeletal Muscle and Exercise Physiology Study Section (SMEP)
Program Officer
Porter, John D
Project Start
2004-09-01
Project End
2010-04-30
Budget Start
2006-05-01
Budget End
2007-04-30
Support Year
3
Fiscal Year
2006
Total Cost
$301,802
Indirect Cost
Name
Tulane University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
053785812
City
New Orleans
State
LA
Country
United States
Zip Code
70118
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Xu, Xueqing; Tsumagari, Koji; Sowden, Janet et al. (2009) DNaseI hypersensitivity at gene-poor, FSH dystrophy-linked 4q35.2. Nucleic Acids Res 37:7381-93

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