PROJECT 4 FSH-Patient Derived IPS Cells to Study the Developmental Regulation of DUX4 Transcription The D4Z4 repeat array with multiple copies of the intronless DUX4 gene likely developed as a consequence of retrotransposition of an ancestral gene expressed in ES cells. Our recent publication and preliminary data shows that transcripts from D4Z4 are readily detectable in ES cells making them an important cell type for the study of DUX4 transcriptional regulation. Our preliminary data are consistent with the idea that DUX4 is expressed constitutively in ES cells and its regulation is largely achieved by transcriptional silencing as cells differentiate into other tissue types. Our hypothesis is that FSHD is caused by sequence variations in D4Z4 that disrupt this silencing process with unique toxicity to muscle cells. Our long term goal is to define the normal and pathogenic transcription patterns during development, and identify sequences responsible for the altered pattern of gene expression in FSHD.
In Aim 1 we describe experiments to develop and characterize IPS cells from controls and FSHD-affected patients, and determine if epigenetic markers of D4Z4 regulation seen in ES cells are recapitulated in IPS cells.
In Aim 2, experiments are described to identify transcriptional enhancer and silencing regions that are presumably the targets of developmental regulation of the D4Z4 locus, and therefore will be important targets for therapeutic intervention. We will determine how transcriptional regulation is altered when these areas are removed or mutated using human and mouse developmental models.
In Aim 3 we will modify cells from FSHD-affected patients to produce isogenic clones with homozygous D4Z4 genotypes and quantify transcription differences that result as a consequence of these modifications. In so doing we will produce myoblasts from FSHD patients that contain only non-pathogenic an-ays and with advances in IPS cell development, could be used for autologous transplantation.

Public Health Relevance

The significance of these studies is that an understanding of normal and pathogenic developmental patterns of DUX4 transcriptional regulation will provide a model for disease pathophysiology and help identify therapeutic targets for treatment.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Program Projects (P01)
Project #
5P01NS069539-04
Application #
8434929
Study Section
National Institute of Neurological Disorders and Stroke Initial Review Group (NSD)
Project Start
Project End
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
4
Fiscal Year
2013
Total Cost
$224,121
Indirect Cost
Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
078200995
City
Seattle
State
WA
Country
United States
Zip Code
98109
van den Boogaard, Marlinde L; Lemmers, Richard J F L; Camaño, Pilar et al. (2016) Double SMCHD1 variants in FSHD2: the synergistic effect of two SMCHD1 variants on D4Z4 hypomethylation and disease penetrance in FSHD2. Eur J Hum Genet 24:78-85
Jagannathan, Sujatha; Shadle, Sean C; Resnick, Rebecca et al. (2016) Model systems of DUX4 expression recapitulate the transcriptional profile of FSHD cells. Hum Mol Genet :
van den Boogaard, Marlinde L; Lemmers, Richard J L F; Balog, Judit et al. (2016) Mutations in DNMT3B Modify Epigenetic Repression of the D4Z4 Repeat and the Penetrance of Facioscapulohumeral Dystrophy. Am J Hum Genet 98:1020-9
Tawil, Rabi; Padberg, George W; Shaw, Dennis W et al. (2016) Clinical trial preparedness in facioscapulohumeral muscular dystrophy: Clinical, tissue, and imaging outcome measures 29-30 May 2015, Rochester, New York. Neuromuscul Disord 26:181-6
Knopp, Paul; Krom, Yvonne D; Banerji, Christopher R S et al. (2016) DUX4 induces a transcriptome more characteristic of a less-differentiated cell state and inhibits myogenesis. J Cell Sci 129:3816-3831
Daxinger, Lucia; Tapscott, Stephen J; van der Maarel, Silvère M (2015) Genetic and epigenetic contributors to FSHD. Curr Opin Genet Dev 33:56-61
Statland, Jeffrey M; Donlin-Smith, Colleen M; Tapscott, Stephen J et al. (2015) Milder phenotype in facioscapulohumeral dystrophy with 7-10 residual D4Z4 repeats. Neurology 85:2147-50
Lim, Jong-Won; Snider, Lauren; Yao, Zizhen et al. (2015) DICER/AGO-dependent epigenetic silencing of D4Z4 repeats enhanced by exogenous siRNA suggests mechanisms and therapies for FSHD. Hum Mol Genet 24:4817-28
Statland, Jeffrey M; Shah, Bharati; Henderson, Don et al. (2015) Muscle pathology grade for facioscapulohumeral muscular dystrophy biopsies. Muscle Nerve 52:521-6
Statland, Jeffrey M; Odrzywolski, Karen J; Shah, Bharati et al. (2015) Immunohistochemical Characterization of Facioscapulohumeral Muscular Dystrophy Muscle Biopsies. J Neuromuscul Dis 2:291-299

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