FSH Society Research, Fellows and Small
Grants Funded
As of May 1, 2008
[ Active and ongoing project numbers are in bold letters ]
Bronfman Class
Grant: FSHS-MB-001
Researcher: Silvere M. van der Maarel, Ph.D.
Institution:
Dept. of Human Genetics
Wassenaarseweg 72
PO
Box 9503
2300
RA Leiden
The
Netherlands
Project Title: “Generation of Transgenic Mouse Models for FSHD.”
$30,000 7/1/1998 - 6/30/1999 Year 1
$30,000 7/1/1999 - 6/30/2000 Year 2
$30,000 3/31/2001 - 2/28/2002 Year 3
Goal: To initiate groundbreaking research to create FSHD animal models.
Grant: FSHS-MB-002
Researcher: Sara T. Winokur, Ph.D.
Institution: 240 D, Medical Sciences I
Department of Biological Chemistry
Project Title: “Analysis of Chromatin Structure and Skeletal Muscle-Specific Gene
Expression in Facioscapulohumeral Muscular Dystrophy.”
$30,000 6/1/1998 - 5/31/1999 Year 1
$30,000 6/1/1999 - 5/31/2000 Year 2
$30,000 6/1/2000 - 5/31/2001 Year 3
Goal: To initiate novel research using Genechip and gene expression technologies to gain
insight into FSHD. Prior to this project, gene expression studies had never been done in
FSHD.
Grant: FSHS-MB-003
Researcher: Denise Figlewicz, Ph.D.
Institution:
Department of Neurology
Project Title: “Expression of genes proximal to the D4Z4 deletions: a quantitative study in
FSHD patients and controls. “
$30,000 1/1/1999 - 12/31/1999 Year 1
$30,000 1/1/2000 - 12/31/2000 Year 2
Goal: To initiate novel research on gene expression and models to study gene expression in
FSHD.
Grant: FSHS-MB-004
Researcher: David J. Picketts, Ph.D.
Institution:
Research Institute
Project Title: “Utilizing an epigenetic approach to identify the FSHD gene.”
$30,000 5/1/1999 - 4/30/2000 Year 1
$30,000 5/1/2000 - 4/30/2001 Year 2
Goal: To initiate novel research on epigenetic features and hyper sensitive sites to understand
chromosomal aspects and models of gene expression on FSHD. Prior to this project, no
work had been done internationally on elucidating the epigenetic aspects of FSHD.
Grant: FSHS-MB-005
Researcher: Davide Gabellini, Ph.D.
Institution: University of
Howard Hughes Medical Institute
Project Title: “Identification and
characterization of a protein interacting with the
repetitive element causally related to facioscapulohumeral muscular dystrophy.”
$30,000 1/1/2000 - 12/31/2000 Year 1
$30,000 1/1/2001 - 12/31/2001 Year 2
$30,000 1/1/2002 - 12/31/2002 Year 3
Goal: To initiate novel research on gene expression/repression, disease models and to gain
insight into D4Z4 functionality and role in FSHD. This work has led to several landmark
publications on gene mis-regulation, gene silencing and repression complexes.
Grant: FSHS-MB-006
Researcher: Fern Tsien, Ph.D. / Melanie Ehrlich, Ph.D.
Institution:
Human
Genetics/SL31
Project Title: “
FSHD
Cells,
$35,000 5/1/2001 - 4/30/2002 Year 1
$35,000 5/1/2002 - 4/30/2003 Year 2
Goal: To initiate novel research on the role of methylation, chromatin structure and other
epigenetic features in FSHD.
Grant: FSHS-MB-007
Researcher: Tonnie Rijkers, Ph.D.
Institution:
Center for Human and Clinical Genetics
Wassenaarseweg 72
PO
Box 9503
2300
RA Leiden
The
Netherlands
Project Title: “Mouse models to study candidate genes and epigenetic causes of FSHD.”
$30,000 2/1/2003 – 1/31/2004 Year 1
$30,000 2/1/2004 – 1/31/2005 Year 2
Goal: To initiate research on genotype/phenotype correlations in successfully created new lines
of animal models of FSHD.
Grant: FSHS-MB-008
Researcher: Cecilia
Ostlund, Ph.D. / Howard Worman,
Ph.D.
Institution:
Departments
of Medicine and Anatomy and Cell Biology
P
& S 10-518
Project Title: “The role of DUX4 in facioscapulohumeral muscular dystrophy."
$30,000 2/1/2003 – 1/31/2004 Year 1
$30,000 2/1/2004 – 1/31/2005 Year 2
Goal: To initiate research on the role of DUX4, DUX4C and to examine the role of the nuclear
envelope, nuclear lamina and nuclear organization in FSHD.
Grant: FSHS-MB-009
Researcher: Alberto Luis Rosa, M.D., Ph.D.
Institution(1):
WSU
Institution(2): Laboratory of Neurogenetics
Institute for Medical Research “Mercedes y Martín Ferreyra”
INIMEC-CONICET,
National Research Council of
B Col. Velez Sarsfield,
5016
–
Project Title: “Role of nuclear localization signal (NLS) and H1/H2 motifs in DUX4-mediated cell
Death."
$43,750
$14,690
Goal: To gain
understanding on the molecular
pathogenesis
of human facioscapulohumeral muscular dystrophy (
DUX4, a putative double homeobox-containing protein encoded by a 3.3 kb polymorphic
tandem repeat(D4Z4), at the locus FSHD1A on the human chromosomal region 4Q35. It
is
hypothesized that abnormal temporal or spatial expression of DUX4
for muscle cells causing FSHD. The study will help identify the mechanism(s) by which
DUX4 causes cell death.
Grant: FSHS-MB-010
Researcher: Richard Lemmers, MSc., Ph.D.
Institution:
Dept. of Human Genetics
Wassenaarseweg 72
PO
Box 9503
2300
RA Leiden
The
Netherlands
Project Title: : “Refinement of the FSHD critical region on 4qA chromosomes."
$35,000
$35,000 6/15/2006 – 6/14/2007 Year 2
$35,000
2/15/2008 – 2/14/2009 Year 3
Goal:
[Provided by applicant]: Facioscapulohumeral muscular dystrophy (FSHD) is the third
most common myopathy with an autosomal dominant mode of inheritance. FSHD is
caused by contraction of the polymorphic D4Z4 repeat in the subtelomere of
chromosome 4q and the exact pathogenic mechanism is still unclear. An identical and
equally polymorphic D4Z4 repeat is localized on chromosome 10, but this has never been
associated with FSHD. Our approach of detailed characterization of FSHD alleles and
translating these observations to disease mechanisms has provided robust mechanistic
insight in FSHD pathogenesis over the past years, including the mechanism of mitotic
D4Z4 instability (Lemmers et al. 2004a) and the recognition of a bi-allelic 4qter variation
(designated 4qA and 4qB) of which only the 4qA allele is associated with FSHD
(Lemmers et al. 2002). Moreover, our laboratory provided direct evidence for a
chromatin modification associated with the contraction of D4Z4 repeats by demonstrating
hypomethylation of D4Z4 in FSHD alleles (van Overveld et al. 2003).
Through our expertise in pulsed-field gel electrophoresis (PFGE)-based FSHD
allele characterization, we have become the international reference center for FSHD
diagnosis with on average 50 referrals of atypical FSHD patients each year and
culminating in a database of>l000 patient and control genotypes for D4Z4 alleles on
chromosomes 4 and 10. Our PFGE-based D4Z4 examination has led to further
refinement of minimal requirement to develop FSHD in several ways including exclusion
of a region of 55 kb proximal to D4Z4 by identification of proximally extended deletions
in typical FSHD patients (Lemmers et al. 2003). Moreover, and novel to this field, our
analysis provides evidence that within an FSHD repeat, not all units are equal suggesting
that intrinsic differences between individual D4Z4 units within one array may be
important for PSEID pathogenesis (Lemmers et al. 2004a).
In the current application I propose to further refine the minimal region
necessary and sufficient to cause FSHD in two ways. First, I will precisely characterize
three novel patients with an unusual FSHD allele. Two of these alleles carry, analogous
to proximally extended deletions, deletions of sequences distal to D4Z4. The third
pathogenic allele is highly unusual, because preliminary data suggest that it is located on
chromosome 10. The analysis of these alleles will be combined by the full
characterization of FSHD and control alleles that display repeal exchanges between
chromosome 4 and 10. Moreover, I will focus on intrinsic sequence differences between
4qA-, 4qB and 10q-derived D4Z4 units, most notably that of the most proximal unit, as
we provided evidence for a linkage disequilibrium (LD) between this D4Z4 unit and the
distal polymorphism 4qA or 4qB (Lemmers et al. 2004a).
I expect that this proposal will generate new and essential information on the
minimal region that is required to develop FSHD. Considering the complexity of the
disease mechanism, further refinement of these elements is essential for a better
understanding of the primary pathogenic pathway and will assist future research
strategies based on candidate gene approaches and development of appropriate cellular
and animal model systems,
Grant: FSHS-MB-011
Researcher: Yi-Wen Chen, D.V.M., Ph.D.
Institution: Assistant Professors of Pediatrics
Center for Genetic Medicine Research
Children’s
Project Title: “Molecular Mechanisms of Muscle Atrophy in facioscapulohumeral muscular dystrophy"
$30,000 1/1/2006 – 12/31/2006 Year 1
Goal: [Provided
by applicant]: In preliminary studies,
we studied 125 whole genome profiles
of muscle biopsies from patients with facioscapulohumeral muscular dystrophy (FSHD),
Duchenne muscular dystrophy, Juvenile dermatomyositis, dysferlin deficiency, Emery-
Dreifuss muscular dystrophy and 8 other disorders, and showed that one gene, paired-like
homeodomain transcription factor 1 (Pitx1), was significantly and specifically up-
regulated in patients with FSHD. Meanwhile, using data from an independent human
study, we showed that the gene was up-regulated in atrophic muscles of patients with
spinal cord injury. To identify molecular pathways regulated by Pitx1 in mature skeletal
muscles, we over-expressed Pitx1 in mouse muscles, and showed that up-regulation of
Pitx1 lead to induction of the ubiquitin-proteasome pathways, including atrogin 1 which
plays a key role in muscle atrophy.
Based on the preliminary data, we hypothesize that transcriptional pathways
perturbed by the contraction of D4Z4 arrays lead to up-regulation of Pitx1, and induction
of atrophy pathway, which plays critical role in the patho-physiology of FSHD. In this
application, we propose to generate and evaluate a conditional muscle-specific Pitx1
transgenic mouse as a potential animal model of human FSHD. The data generated in the
proposed study will provide valuable findings on molecular mechanisms of muscle
atrophy in general, and a potential animal model for studying FSHD.
Grant: FSHS-MB-012
Researcher: Davide Gabellini, Ph.D.
Institution: Howard Hughes Medical Institute
Program in Gene Function and Expression
University of
Lazare Research Building - 6th Floor - Room 660 A
Project Title: “Development of an Animal Model of FSHD."
$37,500 1/1/2006 – 10/31/2006 Year 1
Goal: [Provided by applicant]: Facioscapulohumeral muscular dystrophy (FSHD), the third
most common myopathy, is an autosomal dominant neuromuscular disorder characterized
by progressive weakness and atrophy affecting selective skeletal muscles. The disease
has not been linked to a classical mutation within a protein-coding gene. Instead, FSHD
patients carry deletions of tandem 3.3 kb repeats, termed D4Z4, located on chromosome
4q35.
An incomplete knowledge of the biochemical pathogenesis of FSHD has
hampered the development of effective therapies.
D4Z4 is a repetitive element with heterochromatic features. Recently, we
reported that
inappropriately over-expressed, specifically in FSHD muscle. We found that an element
within D4Z4 behaves as a silencer providing a binding site for a transcriptional
repressing complex. These results suggest a model in which deletion of D4Z4 leads to the
inappropriate transcriptional de-repression of 4q35 genes, resulting in disease.
To identify the gene(s) responsible for FSHD, we generated transgenic mice over-
expressing
develop a pathology with physiological, histological, ultra-structural and molecular
features analogous to those observed in FSHD patients. These include abnormal spinal
curvature, progressive muscular dystrophy, skeletal muscle atrophy, and differential
involvement of muscle types. Moreover, in both FSHD patients and
mice there is no evidence for mitochondrial involvement or alteration of sarcolemmal
integrity. This latter feature distinguishes FSHD from other muscular dystrophies in
which sarcolemmal disruption is the primary pathogenetic mechanism. By contrast, mice
over-expressing two other putative
FSHD-candidate genes,
with regard to both phenotype and muscle histology.
pre-mRNA splicing. We found that in muscle of
pre-mRNAs undergo aberrant alternative splicing. Collectively, our results suggest that
FSHD results from inappropriate
over-expression of
leads to abnormal alternative splicing of specific pre-mRNAs.
Here we propose a detailed study of
the molecular pathogenesis of FSHD by addressing the following questions:
1)
What is the biological role of
FRG1 is unknown.
might regulate the activity of
splicing factors. We plan to identify
partners as a starting point for understanding its biological role.
2)
How does
the role
of RNAs
it regulates in vivo and the use of a mouse model of FSHD for
validation. To address this aim systematically, we will undertake a genome-wide screen
to identify and validate
These studies will provide relevant information to understand the molecular basis
of FSHD that will help in the
development of effective therapeutic strategies.
mice may be used as a preclinical model to test new therapies for FSHD.
Grant: FSHS-MB-013
Researcher: Melanie Ehrlich, Ph.D.
Institution:
Project Title: “Finding the 4q35 FSHD Gene."
$35,000 7/24/2006 – 7/23/2007 Year 1
$35,000 7/24/2007 – 7/23/2008 Year 2
Goal:
[Provided by applicant]: A major obstacle in research on facioscapulohumeral muscular
dystrophy (FSHD) is the uncertainty about the identity of the 4q35 gene whose activity is
directly controlled by a short D4Z4 array on the same chromosome (in cis).
Circumstantial evidence strongly indicates that inappropriate expression of this gene (the
FSHD gene) in certain skeletal muscle cells is caused by having a short D4Z4 array in its
vicinity. Apparently, the inappropriate expression of the FSHD gene causes the painful
and debilitating symptoms of FSHD by altering expression of other genes indirectly.
I propose to use a novel approach for screening for the FSHD gene in the 1-Mb
region proximal to the D4Z4 array on 4q. There are now well proven examples of long-
distance control of human gene expression by DNA elements that have to be on the same
chromosome as the gene they regulate (cis control). My lab will identify by computer
analysis about 100 sequences that might contain the elusive FSHD gene, including many
sequences that would not be identified by current gene search programs as potential
genes. My research group will design ~100 primer-pairs corresponding to 100-200 bp
sequences in these regions and check by in silico analysis and PCR on human-rodent
somatic cell hybrids and human DNA that these DNA primer-pairs work well in PCR and
are unique to human chromosome 4. This broad search will compensate for the major
inadequacies of available gene prediction programs and allow discovery of either a
conventional or an unconventional gene, such as a gene that encodes a regulatory RNA,
but not a protein. My lab will prepare and characterize myoblasts from FSHD and control
patients and fix these cells. They will then be analyzed by quantitative RNA polymerase
II chromatin immunoprecipitation (ChIP) assays, a DNA-based assay for engagement of
the transcription machinery on specific DNA sequences. Our lab will interpret the
resulting ChIP data and then, on FSHD and control myoblasts, my lab will do RT-PCR
analyses to test sequences that are positive for transcription in the ChIP analysis. These
RNA-based assays will be quantitative real-time RT-PCR analyses to compare FSHD and
control samples and end-point RT-PCR analyses that give another level of verification by
visualization of the size of the RT-PCR product. We will do these RNA-based assays to
verify that the regions are transcribed from myoblasts, to determine if we can detect
increased RNA amounts for one or more of these regions in FSHD vs. control myoblasts,
and to test whether candidate FSHD gene sequences are transcribed from various other
cell types, including FSHD and control fibroblasts and lymphoblastoid cell lines.
The method that we will employ to screen for the FSHD gene is the best one for
direct identification of transcription of genes, whatever their nature. It is independent of
secondary factors that can greatly impact standard RNA analyses. These complicating
factors are RNA degradation in vitro despite the use of RNase inhibitors and RNase-free
reagents, RNA processing in vivo, and RNA stability in vivo. If the RNA polymerase II
ChIP assays indicate differential transcription of one or more 4q35 genes in a comparison
of FSHD to control myoblasts but the RNA assays do not, it could be because of one of
these complications associated with RNA analysis. In that case, we will use a different
type of ChIP assay to confirm the RNA polymerase II ChIP results, namely ChIP with an
antibody to the general transcription factor TBP and PCR primers in the region of the
putative promoter. This study holds the promise of greatly facilitating research on FSHD
by elucidating the nature of the critical gene initially impacted by a short 4q D4Z4 array
but unresponsive to a short, virtually identical D4Z4 array at 10q.
Grant: FSHS-MB-014
Researcher: Patrick Reed, Ph.D.
Institution: Department of Physiology
University
of
660
Project Title: “Analysis of Changes in the Proteome in Facioscapulohumeral Muscular Dystrophy."
$30,000 1/24/2007 – 1/23/2008 Year 1
$30,000 2/15/2008 – 2/14/2009 Year 2
Goal:
[Provided by applicant]: One of the largest hurdles to understanding the pathogenic
basis of FSHD is identifying the molecular mechanisms that trigger the onset of muscular
dystrophy. Although FSHD is linked to deletions of 3.3 kb non-coding repeats (D4Z4)
repeats near the telomere of chromosomal position 4q35, the mechanisms linking these
deletions to changes in gene expression, altered protein expression, and consequent
changes in muscle structure and function, are still poorly understood. Many muscular
dystrophies are linked to changes in the stability of the sarcolemma of skeletal muscle.
My previous work suggested that the sarcolemma of FSHD muscle was also affected in
ways that might lead to muscle weakness, but the proteins that were altered could not be
readily identified. I have now adapted a proteomic approach to identify proteins that are
altered in FSHD, and to test their possible roles in pathogenicity.
The method I have chosen is high resolution, large format, two-dimensional
electrophoresis (2D-GE). With the improvements I have introduced into the method, I
can now detect more than 3000 distinct protein spots in normal and FSHD muscle
samples. Remarkably, my preliminary results indicate that very few proteins show
changes in expression levels in FSHD muscle compared to controls. One, a spot that
showed strong expression in the soluble fraction from FSHD muscle but no detectable
expression in controls, has an isoelectric point of 5.07 and a molecular mass of
approximately 34 kDa. I used LC/MS/MS techniques to show that this protein is mu-
crystallin (CRYM; also called “thyroid hormone binding protein" THBP). Western blots
confirmed that this protein is highly up-regulated in deltoid muscle from FSHD patients
compared to controls. Although my analysis is still incomplete, this protein is of
considerable interest because it is expressed in the retina and is responsible for high
frequency hearing loss, both of which are compromised in patients with FSHD.
Furthermore, its role as a thyroid hormone binding protein places it at a potentially
crucial point in the regulation of myoblast cell division and differentiation, which have
recently been implicated as defective in FSHD through gene array studies. It may also be
linked to sarcolemmal and sarcomeric changes, as crystallins are likely to play important
roles in the assembly of intermediate filaments at these structures in developing muscle
Finally, the autosomal dominant nature of FSHD suggests a “gain-of-function" mutation,
consistent with the over-expression of a protein in FSHD that is expressed at much lower
levels in healthy muscle. My novel findings therefore suggest that FSHD may be caused
by up-regulation of CRYM, with consequent changes in the structural organization and
thyroid hormone signaling pathways.
My general aim is to test the idea that up-regulation of CRYM is an important
pathogenic mechanism that leads to FSHD.
My specific aims are: (i) to learn if increased levels of CRYM are indeed
specific for FSHD by applying my improved methods for 2D-GE to complete my
analysis of the proteomes of FSHD and control muscle, as well as muscles from other
dystrophic samples; (ii) to use cellular transfection methods to study the biology of
CRYM in myoblasts and myotubes in culture; and (iii) to use transgenic techniques to try
to reproduce key features of FSHD in mice. If successful, my experiments should lead to
a new understanding of the molecular mechanisms underlying FSHD, and provide an
animal model to use in developing therapies for it.
Grant: FSHS-MB-015
Researcher: Yvonne Meijer-Krom, Ph.D.
Institution:
Department of Human Genetics
Postal Zone S-4-P
Einthovenweg 20; 2333 ZC Leiden
PO
Box 9600; 2300 RC Leiden
The
Project Title: “Towards the Discovery of Early Developmental Defects in FSHD.”
$35,000 1/24/2007 – 1/23/2008 Year 1
$35,000 1/24/2008 – 1/23/2009 Year 2
Goal:
[Provided by applicant]: Autosomal dominant Facioscapulohumeral muscular dystrophy
(FSHD) is the third most common myopathy. FSHD is mainly characterized by an often
asymmetric progressive weakness and wasting of the facial, shoulder and upper arm
muscles, typically starting in the second decade of life. FSHD is caused by contraction of
the polymorphic D4Z4 repeat in the subtelomere of chromosome 4q (van Deutekom et al.
1993; Wijmenga et al. 1992). Contraction of D4Z4 is associated with DNA
hypomethylation (van Overveld et al. 2003) and loss of a D4Z4 repressor complex
containing the polycomb protein YY1 implying a complex epigenetic disease
mechanism.
There is strong clinical evidence that FSHD should be regarded as a congenital
disease with progressive character. Clinical. and genetic features suggest an embryonic
involvement in FSSHD. These include the marked asymmetry of muscle involvement,
the 1,000-fold increased occurrence of pectus excavatum unrelated to the muscle
weakness and the early onset FSHD cases with complete absence of some muscle groups
(Padberg 1982; Padberg 2004). In addition, two interesting candidate genes, FSHD
region gene 1 (FRGI) and FRG2, located on chromosome 4, are transcriptionally
deregulated in FSHD muscle culture, but not in adult muscle. Involvement of an early
myogenic defect in FSHD is further supported by the observation that many of the
deregulated genes in FSHD muscle are direct targets of MyoD, a key regulator of
myogenesis (Figlewicz et al. 2004; Winokur, et al. 2003). Loss of YY1-Ezh2 has been
demonstrated to recruit MyoD, leading to the transcriptional induction of genes involved
in myogenic differentiation (Caretti et al. 2004). Therefore, we hypothesize that an
unbalanced YY1 availability during early embryogenesis disturbs the myogenic program,
which may render specific muscle groups more susceptible to disease later in life.
To obtain better insight in the direct targets of MyoD that are deregulated in
FSHD, we will perform a transcriptome analysis of4q-linked FSHD, phenotypic FSHD
with hypomethylation of D4Z4 and control fibroblast undergoing forced myogenesis. To
determine their dependency on YYl, YY1 levels will he reduced during myogenesis. In
parallel, the fusion and differentiation rate of the fibroblast cell cultures will be evaluated
to assess their morphologic characteristics. In advantage over the assessed gene
expression profile in mature rnuscle, the current application mimics the early myogenic
program. Furthermore, the forced myogenic cell population will be much more
homogeneous compared to primary myoblast cell cultures (Bergstrom et al. 2002; Berkes
et al- 2004; Padberg 1982). We expect this study to provide new and essential
information on the early (embryonic) component of the FSHD phenotype.
Grant: FSHS-MB-016
Researcher: Darko Bosnakovski, D.V.M., Ph.D.
Institution: Perlingeiro lab (ND5.120)
Center for Developmental Biology
UT
Southwestern
Project Title: “Molecular Analyses of DUX4 and Interaction with Myogenic Regulators in FSHD.”
$21,488 12/01/2007 – 11/30/2008 Year 1
Goal:
[Provided by applicant]: The prevailing model for FSHD is that deletion of D4Z4 repeats at
4q35.2 causes local modification of heterochrornatin resulting in deregulation of nearby genes.
Which gene(s) may be directly responsible for FSHD is controversial. In preliminary studies in
which I used an inducible gene expression system to screen FSHD candidate genes, I found
that only DUX4, a candidate located within each D4Z4 repeat, has structural and toxic effects
at a variety of expression levels in C2C12 mouse myoblasts. Furthermore, C2C12 cells induced
to express DUX4 showed striking gene expression similarities to myoblasts from FSHD patients.
Bth display deregulation of MyoD and oxidative stress related genes. In addition, I show that
DUX4 protein is expressed in cultured myoblasts from FSHD patients. We hypothesize that DUX4
plays a role in the pathogenesis or FSHD.
The goals of this proposal are to understand the mechanism of the toxicity of DUX4
and to explore possible therapeutic interventions. Because DUX4 is extremely toxic to myoblasts
a conditional gene expression system needs to be used to study its effects (Aim 1). In the preliminary
study, by using a doxycycline-inducible DUX4 expression system, I found that several crucial
myogenic genes (MyoD, Myf5) are targets of DUX4 (Aim 1). I propose to study the underlying
mechanism of the toxicity of DUX4 in myoblasts as well as in other cell types. I postulate that
appropriate intervention of the deregulated genes (by over-expression or RNAi knockdown) in
DUX4 affected cells should rescue the toxic phenotype (Aim 1). Furthermore I will test the
hypothesis that DUX4 interferes with the function of myogenic regulators by competitive binding
to the same target sites (Aim 2). In support of this, over-expression of Pax3, a crucial gene
in myogenesis and whose homeodomain is most similar to DUX4, renders C2C12 cells resistant to
DUX4-mediated toxicity. The aims of this proposed study target the most crucial and unknown
aspects (both mechanism and therapy) of FSHD.
Aim 1. To understand the mechanism of action of DUX4.
Aim 2. To test the hypothesis that DUX4 interferes with the function of myogenic regulators
Significance
The pathogenic mechanism of FSHD is controversial and largely unknown, which is the
major hurdle in developing a rational therapy. Therefore it is extremely important to find the
gene(s) involved in FSHD, and to understand their action, from which therapeutic strategies
will arise. My proposed study is designed to answer these crucial questions. Using uniform gain
of function approach, I will look closely on the effects of the FSHD candidate gene, DUX4 on
myoblast phenotype, analyze the underlying mechanism in detail (Aim 1) and identify potential
targets in the cascade of pathogenesis (Aim 2). Thus this study is directly relevant to progress
towards a therapy for FSHD.
Grant: FSHS-MB-017
Researcher: Paola Picozzi, Ph.D.
Institution: Stem Cell Research Institute
DIBIT-HSR 1B4-Room 97
Via Olgettina 58
20132 Milano
Project Title: “Functional characterization of D4Z4 in FSHD”
$35,000 3/1/2008 – 2/28/2009 Year 1
Goal:
[Provided by applicant]: The long-term goal of our research is to identify and characterize the
molecular pathways that become subverted in facioscapulohumeral muscular dystrophy (FSHD)
in order to develop therapeutic strategies. FSHD, the third most common myopathy, is an
autosomal dominant neuromuscular disorder characterized by progressive weakness and
atrophy affecting selective skeletal muscles. Unlike the majority of genetic diseases, FSHD
is not caused by a classical mutation within a protein-coding gene but rather involves a
complex cascade of epigenetic events following contraction of a 3.3 kb subtelomeric non-coding
repeat (D4Z4) located on chromosome 4q35.
At present no treatment is available for FSHD. This has been also hindered by an
incomplete knowledge of the disease pathogenesis and, until recently, by the lack of an animal
model. Based upon recent experimental results, it has been proposed that deletion of D4Z4 leads
to the inappropriate transcriptional de-repression of the 4q35 gene FRG1 resulting in disease.
Understanding how deletion of D4Z4 causes up-regulation of 4q35 genes is important to develop
therapeutic approaches aimed at preventing transcriptional de-regulation in FSHD.
Our specific aims are:
1. To characterize protein/DNA interactions at D4Z4. It has been shown that a transcriptional
repressor complex composed of YY1, HMGB2 and nucleolin is associated with D4Z4 (Gabellini et al, 2002).
In mammalian cells, transcriptional repression is the result of the cooperation between sequence specific
repressors and general co-repressors such as histone deacetylases (HDACs) and DNA methylases.
Notably, the activity of YY1 is regulated at the posttranslational level, possibly through interactions
with other proteins. YY1 represses transcription by interacting with HDAC-1 and 2, and this interaction
is regulated by HDAC phosphorylation. (Galasinski et al, 2002). Collectively, these observations
suggest that other proteins may be associated with and regulate the activity of the DRC.
2. To elucidate the mechanism underlying control of gene expression at 4q35.
Our preliminary results suggest that non-coding RNAs and microRNAs generated by D4Z4 regulate
chromatin structure and 4q35 genes expression. Our analysis will generate novel insights into the
biological role of repetitive DNA sequences in higher eukaryotes.
The results of these studies will be very useful to identify effective therapeutic approaches for FSHD.
Delta Railroad
Construction Class
Grant: FSHS-DR-001
Researcher: Alexandra Belayew, Ph.D. / Stephane Plaisance, Ph.D.
Institution: Lab. Biologie
Moléculaire
Université
de Mons-Hainaut
Pentagone, avenue du Champ de Mars 6
B
- 7000 -
Project Title: “Characterization of a protein expressed from a 3.3 kb element not linked to FSHD.”
$15,000 6/1/1998 - 12/31/1998 Year 1
“Small
laboratory equipment for research on FSHD."
$15,000 2/15/2001 - open Year 1
Goal: To initiate research on the role of DUX, DUX1, DUX4, DUX4C and to elucidate the role
of DUX in FSHD and within the D4Z4 region.
Grant: FSHS-DR-002
Researcher: Rossella Tupler, M.D., Ph.D.
Institution: Howard Hughes Medical Institute,
University
of
Project Title: “Characterization of differentially expressed genes in facioscapulohumeral
muscular dystrophy affected muscles.”
$30,000 6/1/1998 - 5/31/1999 Year 1
Goal: To initiate research into differentially expressed genes involved in FSHD. This
groundbreaking research has led to major advances in our understanding of FSHD.
Grant: FSHS-DR-003
Researcher: Robert Bloch, Ph.D.
Institution:
660
Project Title: “Sarcolemmal organization in FSHD and the MYD mouse.“
$30,000 7/1//1999 - 4/30/2001 Year 1 (interrupt/extend)
Goal: To gain insight into structural aspects and patho-physiology of FSHD using the latest
techniques as well as revisiting standard methodologies. To examine the structure of
FSHD muscle and the sarcolemma for insights into the disease.
Grant: FSHS-DR-004
Researcher: Jane Hewitt, Ph.D.
Institution:
Division
of Genetics
Queen’s
Medical Centre
NG7
2UH,
Project Title: “Fugu rubripes as a model organism for FSHD gene identification.”
$30,000 7/1/2000 - 6/30/2001 Year 1
Goal: To sequence the analogous 4q35 region in puffer fish for insight into
genomic organization of FSHD. To use data to help with mapping, assembly and
finishing of the 4q35 human region. Based on this research, we were able to assist the
Human
Genome Project at
very difficult and recalcitrant region of 4q35.
Grant: FSHS-DR-005
Researcher: Marcy Speer, Ph.D.
Institution:
Project Title: “Genetic Linkage Studies in Non-chromosome 4 FSHD.”
$30,000
Goal: To examine and find the genetic locus of the non-chromosome 4 families through
genome wide search/scan. This project is a high priority for the research community. It
aims to register non-chromosome 4 pedigrees with the researchers and clinicians at Duke.
The
Grant: FSHS-DR-006A
Researcher: Emma Ciafaloni, M.D
Institution:
Department of Neurology
Project Title: “The
Course and Outcome of Pregnancy and Delivery in Women with
Dystrophy.”
$13,074
$1,926
$0
Goal: Very
little is known about the course
with muscular dystrophies. Our current ability to efficiently counsel women with
muscular dystrophies when pregnant or planning a pregnancy is very limited due to the
lack
of studies addressing the issue of pregnancy
specific
attention
progression of the myopathy. Objectives are: to increase our knowledge about the course
and
outcome of pregnancy
assess
the effect of pregnancy, delivery
weakness
and,
to ultimately improve counseling, family planning
women
with
Grant: FSHS-DR-006B
Honoraria
Researcher1: Wendy M. King, PT
Institution1:
389 McCampbell Hall
Researcher2:
Institution2:
Physical Medicine and Rehabilitation
Project Title: “Facioscapulohumeral
muscular dystrophy Physical Therapy Booklet/Brochure
Article for Physical Therapy Journal.”
$15,000
Goal: Gather and review of literature/information related to FSHD natural history, surgical
options, orthotics, rehabilitation, physical therapy interventions, role of exercise,
hydrotherapy,
pain, etc. Review scientific literature,
brochures
organizations
from English speaking countries to assess the type
information already available. Draft, peer-review and publish booklet/brochure on FSHD
and Physical Therapy and submit journal article to Physical Therapy journal on P.T. and
FSHD.
Grant: FSHS-DR-007
Researcher: Sara Winokur, Ph.D. / Ulla Bengtsson, Ph.D.
Institution: 202 Sprague Hall
Biological
Chemistry
University
of
Project Title: “Coding and non-coding RNA expression in FSHD.”
$35,000
Goal:
[Provided by applicant]: More than a decade after the position effect hypothesis was first
proposed, the fundamental question of whether altered chromatin structure in FSHD
affects RNA expression at 4q35 has not be answered. Several independent laboratories
have addressed this question, yielding disparate and contradictory results. In part, this is
due to the variability in tissues and cultures utilized by various laboratories, which are
provided by different sources and often obtained and preserved using different methods.
In addition, all of the experimental techniques used to examine RNA expression thus far
have relied on pooled sources of RNA from tissues or cell cultures. These techniques
include non-quantitative RT-
studies assayed differential RNA expression between FSHD and control muscle, and, by
nature of the experimental design, detected average RNA levels emanating from both
alleles and multiple cell types.
In contrast, examination of RNA expression in a single cell context is more
suited to address the question of whether an altered chromatin structure on the contracted
D4Z4 allele influences RNA expression. RNA-FISH (fluorescence in situ hybridization)
utilizes antisense RNA or dsDNA as hybridization probes to nascent nuclear RNA
transcripts followed by fluorescence detection of conjugated haptens or antibodies.
Transcription of both coding and non-coding RNAs from each of the alleles (normal and
D4Z4 contracted) can be readily identified by RNA-FISH followed by hybridization with
D4Z4 and 4q specific
can be readily identified using this technique, either in culture or within tissue sections.
We
propose to utilize
4q35 genes are transcribed in proliferating myoblasts and differentiated myotubes? 2)
Are the levels of transcription different between normal and FSHD myoblasts/myotubes?
3) Is there an allele specific transcription in FSHD myoblasts/myotubes? That is, do the
contacted and normal alleles
display different levels of
cells? For these studies, 3’ hyper-biotinylated antisense oligos corresponding to 4q35
genes will be used as probes for
coding
myotubes.
If chromatin structure is altered in FSHD, leading to aberrant RNA expression,
then we should not assume that such a mechanism would affect coding RNA exclusively.
Non-coding RNA has increasingly come to light as a significant player in the regulation
of both transcription and translation. Although several approaches to the detection of
non-coding RNAs exist , we propose to use the same technique (RNA-FISH) to examine
non-coding RNA within a defined region proximal to the D4Z4 repeat. Genomic clones
(cosmids) will be used to hybridize to these RNAs as the specific non-coding transcripts
cannot be identified a priori.
Lastly, RNA transcription of genes affected in FSHD (as identified by
expression profiling) will be examined in FSHD and control myoblasts/myotubes. A
recent finding in FSHD research within the past year has been the unique and consistent
localization of the 4q telomeric region to the nuclear periphery. While the biological
significance of this localization is not yet known, the existence of nuclear domains either
permissive or repressive of transcription is well documented. Therefore, genes affected in
FSHD will be examined by RNA-FISH to determine whether co-localization with the
FSHD region at the nuclear periphery might affect RNA transcription from these genes.
Grant: FSHS-DR-008
Researcher: Jane Hewitt, Ph.D.
Institution:
Queen's Medical Centre
NG7 2UH
Project Title: “Development of Genomic Resources for Functional Studies of the Mouse DUX4 Array
in Vivo.”
$29,658 7/24/2006 – 7/23/2007 Year 1
$15,000 11/1/2007 – 5/31/2008 Year 2
Goal:
[Provided by applicant]: We have recently demonstrated conservation of the open
reading frame and the tandem array organization of Dux4 homologues in a wide range of
mammalian species, suggesting a protein- encoding function for the array and a
requirement for a high copy number. We hypothesize that the conservation of the open
reading frame and the tandem array organization of DUX4 homologues in a wide range
of mammalian species indicates a protein-encoding function for the array and a
requirement for a high copy number. This function may be disrupted by the FSHD
deletion and hence play a role in the disease mechanism. The identification of the mouse
homologue (Dux4) provides a model organism in which to genetically manipulate the
Dux4 array in vivo. In the work proposed in this application we plan to generate a set of
resources that will then enable us to generate of mouse lines that either a) have reduced
repeat numbers within the Dux4 arrays or b) in which the entire Dux4 array is deleted. In
specific aims 1 and 2 we will complete the physical and the sequence map of this locus.
In specific aim 3, using information from this region obtained in aims 1 and 2, we will
generate gene targeting constructs using the Mutagenic Insertion and Chromosome
Engineering Resource (MICER),
developed in the
FSH Society Landsman Charitable Trust Fellowship
Grant: FSHS-LCT-001
Researcher: Meredith Hanel, Ph.D.
Institution: Department of Cell and Developmental Biology
B107 Chemical and Life Sciences Laboratory
Project Title: “An in vivo Xenopus System for Studying D4Z4 Mediated Chromatin and Gene Expression”
$30,000 03/01/2008 – 2/28/2009 Year 1
Goal:
[Provided by applicant]: We have created a novel animal model exhibiting an FSHD
phenotype using Xenopus laevis frogs, supporting the assertion that FSHD pathology is
due to the over-expression specific genes. This provides two therapeutic targets; 1) the misregulation
of the gene by the mutated D4Z4 array and 2) the activity of the over-expressed gene product. This
proposal addresses the former, seeking to understand the regulation mediated by the D4Z4 array and
4q subtelomere by systematically recapitulating the human FSHD region of chromosome 4q35 in the frog.
Introducing these human sequences, and particularly the D4Z4 repeats, under controlled circumstances
will allow us to determine the regulatory requirements lacking in the FSHD deletion.
Xenopus transgenesis enables monitoring integrated genomic copies of engineered
sequences in a developing animal in a high through-put method. The telomeric nature of the
environment of the D4Z4 repeats will even be recapitulated in our system as we have recently
engineered telomeres in Xenopus (Wuebbles and Jones, 2007). The Xenopus genome contains many
of the same epigenetic characteristics as humans, making potential findings applicable to humans. The goal
of this proposal is to determine the cis and trans regulatory requirements for normal gene repression in the
4q35 region and how this is disrupted in FSHD.
Our specific aims are:
Aim 1: The impact of D4Z4 repeat number on the expression of FRG1 and neighbouring
genes. D4Z4 repeats will be placed in cis with human and Xenopus FRG1 promoters driving the reporter GFP.
Cis effects on gene expression will be visualized in a developing vertebrate and correlated with number
of repeats. Subsequent analysis of DNA methylation and chromatin structure will determine the
nature of repression or activation. Experiments will test the major hypotheses in the field that (1) D4Z4
repeat mediated gene repression is due to a local repressive effect of heterochromatin spreading,
(2) A repressor bound to D4Z4 repeats associates with promoters, (3) An activator associated with
the D4Z4 repeats activates transcription.
Aim 2: Regulatory roles of the subtelomere region. The repetitive nature of CpG rich D4Z4
repeats and their subtelomeric location suggest a structural role or maintenance of chromatin
conformation. D4Z4 repeats have been proposed as an insulator from or propagator of telomere
position effects. Since FSHD is strictly associated with the 4qA allele (containing a distal Beta satellite
repeat), but not the 4qB allele (without beta satellites) telomeric transgenes containing D4Z4 arrays
between the telomere and a reporter gene will test the influence of Beta-satellite DNA on gene
expression as well as the insulator activity of the D4Z4. The number of D4Z4 repeats required to
overcome telomeric position effect and the effect of the Beta-satellite DNA will be assayed. To
test whether D4Z4 propagate heterochromatin we will assess the ability of D4Z4 repeats to override
the effect of the Beta-globin HS4 insulators and result in gene repression.
Aim 3: D4Z4 repeats as transcriptional regulators with genome wide effects. If D4Z4 repeats
regulate genes in trans, adding D4Z4 repeats to the Xenopus laevis genome may sequester proteins that
bind D4Z4, and may ultimately manifest as a developmental phenotype. Since Xenopus development
occurs externally, embryos at any stage of development are easily visualized and alterations in candidate
myogenic markers and vasculature will be assayed.
Xenopus provides a unique opportunity to observe developmental stage and tissue specific
differences in epigenetic and gene regulation. Combined with the intense research by numerous groups
into pharmaceuticals targeting epigenetic regulators, elucidating the factors involved in gene regulation
by the 4q35 region may make these treatments applicable to FSHD.
Grant: FSHS-LCT-002
Researcher: Scott Q. Harper, Ph.D.
Institution: Center for Gene Therapy
The
700 Children's Drive, Room WA2015
Project Title: “In Vivo Investigation of DUX4 As A Candidate FSHD Gene”
$10,000 03/01/2008 – 2/28/2009 Year 1
Goal:
Provided by applicant]: Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal
dominant disorder characterized by progressive and asymmetric weakness of facial, shoulder, and limb
muscles. It is the third most common muscular dystrophy and no effective treatment exists. FSHD is
caused by contraction of D4Z4 repeats on human chromosome 4q35. Though the causative mutation
has been known for nearly 15 years, the underlying pathogenic mechanism for the disease remains
unresolved. Current models suggest that normal chromatin structure at 4q35 is altered by pathogenic
D4Z4 arrays (1-10 repeats) leading to aberrant up-regulation of nearby genes. To date, FRG1 is arguably
the best candidate FSHD candidate gene; FRG1 over-expression in mice recapitulates some dystrophic
changes associated with FSHD but the gene is not uniformly elevated in all patient biopsies. Thus, its
uncertain role in FSHD pathogenesis justifies the search for other candidates. Recent evidence suggests
DUX4 may play a role in FSHD development. DUX4 is the translated product of a transcript arising from
D4Z4 which induces apoptosis of cultured myoblasts upon over-expression. However, the in vivo effects
of DUX4 over-expression in muscle are unknown. The goal of this project is to determine whether viral
vector-mediated DUX4 over-expression in mouse muscle causes histological changes associated with
FSHD. This work will be an important step toward understanding the pathobiology of FSHD, which is
necessary for ultimately developing effective therapies.
Specific Aim: To investigate the in vivo effects of DUX4 over-expression in muscle. DUX4 is
candidate FSHD gene due to its chromosomal location (as a product of D4Z4 repeats) and because its over-
expression induces apoptosis in cultured myoblasts. In vivo DUX4 over-expression in muscle is a logical
next step toward investigating its potential role in FSHD pathogenesis. Adeno-associated viral (AAV) vectors
are ideally suited for in vivo muscle gene delivery because they efficiently transduce muscle at high levels,
produce no adverse effects on muscle histology/physiology, and allow cheap and rapid analysis compared
to transgenic mouse methods. Here, we will use AAV serotype 8 (AAV8) vectors to deliver DUX4 or
control genes to muscles preferentially or minimally affected in FSHD. We hypothesize that DUX4
over-expression will induce histological changes associated with muscular dystrophy in transduced animals.
This study will be an important first step toward understanding the potential role of DUX4 in FSHD
pathogenesis and may have future implications for developing FSHD therapies.
Grant: FSHS-TF-001
Researcher: Marcy Speer, Ph.D.
Institution:
Project Title: “Genetic Linkage Studies in Non-chromosome 4 FSHD.”
$30,000 2/1/2001 - 1/31/2002 Year 1 (See Year 2 under Delta RR)
Goal: To ensure that the work to examine and find the genetic locus of the non-chromosome 4
families continues. This project is of very high priority to the FSH Scientific Advisory
Board.
Grant: FSHS-VR-001
Researcher: Robert Bloch, Ph.D.
Institution:
660
Project 1 Title: “Sarcolemmal organization in FSHD and the MYD mouse.“
$15,000 5/1//2001 -
Project 2 Title: "To investigate the “proteome” in FSHD and to compare it to the “proteome” in
control muscles and in other common myopathies and muscular dystrophies
using
Two-dimensional gel electrophoresis"
$15,000 2/15//2002 - 8/15/2002 Year 1
Goal: To gain insight into structural aspects and patho-physiology of FSHD using the latest
techniques as well as revisiting standard methodologies. To newly examine the structure
of FSHD muscle and the sarcolemma and to examine more closely the proteins involved
in FSHD using proteomic approaches.
Grant: FSHS-TG-001
Researcher: Jeanne Lawrence, Ph.D. / Y. Polly Xing, M.D., Ph.D.
Institution: University of
Project Title: “Higher level chromatin packaging and nuclear organization of FSHD cell with
an emphasis on its 3.3 kb deletion involving high resolution transcript mapping
by mRNA in situ and direct visualization of this region of the chromosome via
In situ hybridization with loop halo DNA preparations."
$30,000
Goal: To gain insight into nuclear organization, scaffolding, structure and chromatin packaging
involved in FSHD. To examine epigenetic features, nuclear compartmentalization and
aspects of D4Z4 and telomere organization.
Grant: FSHS-SMRF-001
Researcher: Graham J Kemp, M.D.
Institution: Faculty of Medicine
Project Title: “Muscle damage by reactive oxygen species, muscle atrophy and effects of creatine
supplementation in facioscapulohumeral muscular dystrophy."
$35,000
($48,650 total see balance under Lewis)
Goal: This a pilot study designed to test the following hypotheses: (1 that muscle in FSHD
shows
evidence of damage by
anti-
that this also partially alleviates muscle atrophy, even in the absence of training, and; 5)
that
this results in an increase in muscle strength
label
patients
with proven FSHD.
biopsies
of deltoid. Muscle atrophy
by
whole-body quantitative magnetic resonance imaging (
effects on symptomatology will be be quantified. We will compare pre-creatine results
with
those of control subjects,
values.
This
study
usefulness
of creatine over a longer time
mechanisms
of muscle damage in FSHD; if
compounds
that reduce oxidative stress in muscle
help
in the design
Grant: FSHS-SMRF-002
Researcher: Sara Winokur, Ph.D. / Ulla Bengtsson, Ph.D.
Institution: 202 Sprague Hall
Biological Chemistry
Project Title: “Restoration of normal myogenic pattern in FSHD: A nutritional approach."
$30,000 3/1/2003 – 2/29/2004 Year 1
$30,000 3/1/2004 – 2/28/2005 Year 2
Goal: A clinically oriented project to study patterns of FSHD myogenesis in cell systems using
compounds and nutritional agents that affect methylation, oxidative stress, chromatin
structure and muscle cell differentiation. A major goal of this project is to build an
effective model system to assay target compounds effectively. The objective of this
study is to identify therapeutic compounds to treat FSHD that can be taken as part of a
nutritional regimen. Nutritional compounds are selected based on functional impact on
myogenesis, availability as nutritional supplement and expediency for clinical trials.
Grant: FSHS-SMRF-003
Researcher:
Institution: Head Biomedical Magnetic Resonance group
Department of Radiology (667)
6500 HB
The
Project Title: “Assessment of the metabolic inter-muscular heterogeneity, and muscular creatine uptake
and
turnover in
$30,000 8/14/2006 – 8/13/2007 Year 1
$15,000 8/13/2007 – 2/14/2008 Year 2
Goal:
[Provided by applicant]: Although substantial progress has been made in the molecular
biology of facioscapulohumeral muscular dystrophy (FSHD), little is still known about its
pathophysiology such as possible defects in skeletal energy metabolism. Asymmetric
dys-functioning of muscles is a typical feature of FSHD but characteristic metabolic
profiles of the affected muscles are lacking, and objective biomarkers to assess therapies,
e.g. creatine supplementation, which possibly has beneficial effects, are not available.
MR spectroscopy (MRS) is an ideal tool to study metabolism in muscle in a non-invasive
way.
Hypothesis. The application of MRS to FSHD patients will uncover metabolic
abnormalities that can serve as non-invasive biomarkers to assess, and better understand
the severity of disease in specific muscles. The signals of creatine can serve as non-
invasive biomarkers to assess creatine uptake, phosphorylation and turnover in skeletal
muscle of patients in creatine supplementation treatment.
Study objectives:
(1). To discover metabolic abnormalities in skeletal muscle of FSHD patients by MRS as
biomarkers for the severity of the disease in specific muscles.
(2).To determine if the level of Cr and PCr, by quantitative MRS is decreased in muscle
of FSHD patients
(3). To determine creatine uptake, phosphorylation and turnover in different skeletal
muscles in healthy volunteers (which is not known), and,
(4). in the muscles of FSHD patients.
Study design will consist of two parts.
(I). A metabolic profile of muscles will be assessed using phosphorous 31 (31P) and
tritium (1H) MRS and possible differences between affected and non-Affected skeletal
muscles will be studied (objective I). Specifically, differences in Cr and PCr
concentrations will be monitored (objective 2). Simultaneously, Cr turnover and PCr/Cr
ratios after Cr supplementation will be studied in healthy volunteers (objective 3).
(II). Depending on the results of the volunteer studies Cr uptake, phosphorylation and
turnover will he assessed in a single location or in several skeletal muscles of FSHD
patients to meet objective 4.
Expected results A metabolic profile will be established to serve as non-
invasive biomarker for the severity of disease in specific muscles and to monitor therapy
in FSHD. Differences in Cr uptake and turnover between skeletal muscles in healthy
persons and FSHD will guide the optimization of Cr supplementation strategies in FSHD
patients.
Grant: FSHS-NYSS-001
Researcher: Daniela M. Oliveira, Ph.D.
Institution: Ottawa Health Research Institute
Project Title: “Identification of the mechanism regulating the Wnt-dependent activation of muscle
progenitor cells.”
$30,000
Goal: The overall goal of the project is to identify genes regulated by the Wnt signaling
pathway
that
CD45+/Sca-1+
muscle cells. In addition muscle
satellite cells, another
population within muscle (CD45+/Sca-1+ muscle cells) plays a physiological role in
muscle
regeneration. Identification of
stimulate
the Wnt-target genes that might be used to enhance
FSHD.
Grant: FSHS-NYSS-002
Researcher:
Institution: Department of HumAn Genetics
Project Title: “Testing whether D4Z4 Perform Long Distance Gene Silencing via the Chromosome 4
Inactivation Network.”
$22,652
Goal: A high risk and novel approach to understanding chromosome interactions, epigenetics,
To test the hypothesis that long repetitive sequence on a chromosome, regardless of
sequence, is tied into the network of long repeats responsible for chromosome
inactivation and particular with FSHD the case of non-random mono-allelic autosomal
inactivation. To test the hypothesis that the tract of D4Z4 repeats at 4q35 is tied into the
chromosome
4 inactivation network
inactivation
resulting in abnormal
Grant: FSHS-FS-001
Researcher: Nieves Embade, Ph.D. /
Institution: Department of Human Genetics
Project Title: “Tethering Adenine (Dam) Methylase to the 3.3-kb FSHD Repeats to Identify Distant
Genes that Physically Come in Contact with the Repeats.”
$30,000
Goal: A high risk and novel approach to understanding chromosome interactions, epigenetics,
gene expression in FSHD and with which other parts of the chromosome(s) the FSHD
chromosome 4 D4Z4 repeats are coming into contact. To locate the FSHD gene(s) that
interact with the D4Z4 repeats by tethering bacterial adenine methylase to sequences in
or near the 3.3 kb repeats and then identifying adenine-methylation at distant sites on the
same chromosome and/or other chromosomes.
Grant: FSHS-FS-003
Researcher: Sara T. Winokur, Ph.D.
Institution: 202 Sprague Hall
Biological Chemistry
Project Title: “FSHD nuclear organization and RNA expression in early development.”
$38,000 1/24/2007 – 1/23/2008 Year 1
Goal:
[Provided by applicant]: The precise mechanism responsible FSHD muscular dystrophy
continues to evade elucidation using current approaches to the disease. New resources
and avenues of research are necessary to provide fresh insight and perspective into this
most challenging disease. To this end, we propose to examine gene expression and
nuclear organization in FSHD at the earliest stages of development. Even though typical
age of phenotypic onset in FSHD is during adolescence, there are many indications that
the disease is influenced by early developmental processes. Resources currently utilized
for FSHD research are generated from adult tissue (skin, muscle, blood), as well as stem
cell populations within these tissues (myoblasts, lymphoblasts). However, even these
adult stem cells are committed to specific lineages, and may well not reveal information
regarding the FSHD genome at the early in development. We propose to investigate
FSHD region molecular and cell biology in embryonic stem cells (ESC) as a resource for
the FSHD research community. This study will focus on FSPID region organization and
gene expression in normal development, and will provide a basis for comparison to
altered expression and localization in FSHD cells once they become available.
Grant: FSHS-FS-004
Researcher: Alexandra Belayew, Ph.D.
Institution: Lab. Biologie Moléculaire
Université de Mons-Hainaut
Pentagone 3A, Avenue du Champ de Mars, 6
B-7000
Project Title: “Study of DUX4 mRNA and Protein Expression in FSHD.”
$30,000 1/24/2007 – 1/23/2008 Year 1
Goal:
[Provided by applicant]: In this research proposal, we want to focus on expression of the
DUX4 gene we mapped in each unit of the D4Z4 repeat array that is contracted in FSHD.
The gene was identified several years ago, but semostration of its expression in patient
muscles proved technically very challenging because of its low level, its toxicity, and its
homology to hundreds of DUX genes unlinked to FSHD. We could demonstrate
expression in myoblasts and biopsies of the homologous non-toxic DUX4c protein
encoded by an isolated D4Z4 element 42 kb centromeric of the repeat array. We have
recently beat able to develop very sensitive and specific tools and procedures to detect
DUX4 expression at the mRNA and protein level. In our mRNA studies we detected 2
introns downstream from the D4Z4 stop codon: their occurrence allowed unambiguous
identification of RT-PCR products as bona fide mRNA (not genomic DNA) copies in 4
FSHD myoblast lines but not 3 controls. We raised a monoclonal antibody against the
DUX4 carloxyl-terminal domain that specifically detects the DUX4 (52 kDa) and
homologous DUX4c (47 kDa) proteins on Western blots performed with extracts of cells
transfected with p-CI-neo-DUX expression vectors. The Western blot sensitivity was
recently increased about 20-fold by use of a new peroxydase substrate (Pierce) and
allowed DUX4 dectection in 4 additional FSHD myoblast lines provided by Dr. D.
Figlewicz.
(
J. Mercier (INSERM.
(1) With these tools, our first aim is to evaluate DUX4 mRNA and protein
expression in additional myoblast lines and in muscle biopsies of patients with FSHD and
different D4Z4 copy numbers, or FSHD not linked to 4q365. as well as in controls and
other neuromuscular disorders. Biopsies will be provided by Drs. D. Laoudj.Chenivesse
and J. Mercier as well as by Dr.
P. Lunt (United Bristol Healthcare NHS Trust,
myoblast tines established from muscle biopsies have been provided by Dr. D. Figlewicz
and additional ones wil1 be by Drs. D. Laoudj-Chenivesse and J. Mercier.
(2) Our second aim is based on the observation that the DUX4 mRNA 3' ends
we detected mapped outside of the D4Z4 repeat array. This region differs between the
chromosome 4qA allele and the 4qB one that was never found associated with FSHD.
We want to evaluate whether such DUX4 mRNA's might also be produced from the
chromosome 4qB allele.
In conclusion. we expect these studies to demonstrate whether there is a
correlation between DUX4 gene or protein expression and the presence of the FSHD
phenotype or not.
Grant: FSHS-FS-005
Researcher: Patricia Arashiro, B.Sc., Mayana Zatz, MSc., Ph.D.
Institution: Universidade de São Paulo
Instituto de Biociências
Centro de Estudos do Genoma Humano
Departamento de Genética e Biologia Evolutiva
R. Matão, 277 - sala 211
05508-900
Project Title: “Clinical Variability in Patients Affected by Facioscapulohumeral Muscular Dystrophy (FSHD)."
$15,000 3/1/2007 – 3/1/2008 Year 1
Goal:
[Provided by applicant]: Previous studies from our and other groups have shown that
usually males are on average more often and more severely affected than females, with approximately
20% of patients becoming wheelchair-bound (Padberg et al, 1991; Zatz et al, 1998).
We have previously observed in Brazilian FSHD families that asymptomatic carriers
are present in about 30% of the families and some genealogies seem to concentrate more non-
penetrant cases (Tonini et al, 2004). This observation is in accordance with van der Maarel et al (2000)
who have also observed a female predominance of mosaic asymptomatic carriers.
A remarkable but often neglected observation in many families and populations is the
occurrence of elderly individuals who inherit disease genes but who nevertheless remain healthy
(Nadeau, 2006). The tendency for health to persist despite the presence of susceptibility genes has
several explanations, including modifier genes and protective alleles that confer genetic resistance
to disease (Nadeau, 2001).
The purpose of the present proposal is to look for modifying genes or mechanisms involved
in protecting some individuals against the deleterious effect of the FSHD deletion. Understanding this
mechanism may help us to develop new tools for prognosis of the disease and also for future treatment.
In order to compare the gene expression in patients with discordant phenotypes we are currently
collecting muscle and skin samples from families with clinically affected and asymptomatic carriers. Total
RNA will be isolated from muscle (biceps/ deltoid) tissue using TRIzol (Invitrogen) method and their
quality verified using gel electrophoresis and spectrophotometry. Sample handling and microarray
hybridizations will be done in collaboration with Dr. Louis Kunkel (Biological and Biomedical Science,
Affymetrix MAS 5.0 software and custom software (http://db.chip.org) will be used for initial data
processing, noise analysis, and quality control. Data analysis will also be done in collaboration with
Dr. Louis Kunkel
at the Children’s Hospital in
Transcript analysis offers many technical advantages over protein analysis in that the mRNA
molecules possess high affinity and specificity binding partners. Additionally, mRNA molecules exhibit
equivalent biochemical properties and can be amplified. Moreover, proteomics deal with unavoidable
problems of limited and variable sample material, sample degradation, vast dynamic range (more than 106-
fold for protein abundance alone), developmental and temporal specificity, and disease and drug
perturbations (Tyers, 2003). Other aspects that must be considered are that many signaling and regulatory
proteins are present in the cell at very low levels; only a small percentage of proteins are soluble and are
expressed
at a level compatible with structural analysis (
products are poorly soluble membrane proteins of considerable functional importance (van Regenmortel, 2001).
The collection of informative FSHD families, in whom we have identified symptomatic and unaffected
members (asymptomatic carriers and non-carriers) who are willing to be submitted to a muscle biopsy for
research purposes (after informed consent), is extremely difficult in practice. It has been possible due to
many years of research from our group. In addition, the comparison of gene expression from asymptomatic
carriers and affected patients is a novel approach that might bring important results.
Grant: FSHS-LEWI-001
Researcher: Graham J Kemp, M.D.
Institution: Faculty of Medicine
Project Title: “Muscle damage by reactive oxygen species, muscle atrophy and effects of
creatine supplementation in facioscapulohumeral muscular dystrophy."
$13,650
($48,650 total see balance under Roberts)
Goal: This a pilot study designed to test the following hypotheses: (1 that muscle in FSHD
shows
evidence of damage by
anti-
that this also partially alleviates muscle atrophy, even in the absence of training, and; 5)
that
this results in an increase in muscle strength
label
patients
with proven FSHD.
biopsies
of deltoid. Muscle atrophy
by
whole-body quantitative magnetic resonance imaging (
effects on symptomatology will be be quantified. We will compare pre-creatine results
with
those of control subjects,
values.
This
study
usefulness
of creatine over a longer time
mechanisms
of muscle damage in FSHD; if
compounds
that reduce oxidative stress in muscle
help
in the design
Grant: FSHS-LEWI-002
Researcher: Emma Ciafaloni, M.D
Institution:
Department of Neurology
Project Title: “The
Course and Outcome of Pregnancy and Delivery in Women with
Dystrophy.”
$0
$11,047
$13,363
Goal: Very
little is known about the course
with muscular dystrophies. Our current ability to efficiently counsel women with
muscular dystrophies when pregnant or planning a pregnancy is very limited due to the
lack
of studies addressing the issue of pregnancy
specific
attention
progression of the myopathy. Objectives are: to increase our knowledge about the course
and
outcome of pregnancy
assess
the effect of pregnancy, delivery
weakness
and,
to ultimately improve counseling, family planning
women
with
Grant: FSHS-LEWI-003
Researcher: Sara Winokur, Ph.D. / Ulla Bengtsson, Ph.D.
Institution: 202 Sprague Hall
Biological Chemistry
Project Title: “Coding and non-coding
$30,000 3/1/2006 - Bridge Fund Year 1
Goal:
[Provided by applicant]: More than a decade after the position effect hypothesis was first
proposed, the fundamental question of whether altered chromatin structure in FSHD
affects
have addressed this question, yielding disparate and contradictory results. In part, this is
due to the variability in tissues and cultures utilized by various laboratories, which are
provided by different sources and often obtained and preserved using different methods.
In addition, all of the
experimental techniques used to examine
have relied on pooled sources of
include non-quantitative RT-
studies assayed differential
nature of the experimental design,
detected average
alleles and multiple cell types.
In
contrast, examination of
suited to address the question of whether an altered chromatin structure on the contracted
D4Z4 allele influences
utilizes antisense
transcripts followed by fluorescence detection of conjugated haptens or antibodies.
Transcription of both coding and non-coding RNAs from each of the alleles (normal and
D4Z4 contracted) can be readily
identified by
D4Z4 and 4q specific
can be readily identified using this technique, either in culture or within tissue sections.
We
propose to utilize
4q35 genes are transcribed in proliferating myoblasts and differentiated myotubes? 2)
Are the levels of transcription different between normal and FSHD myoblasts/myotubes?
3) Is there an allele specific transcription in FSHD myoblasts/myotubes? That is, do the
contacted and normal alleles
display different levels of
cells? For these studies, 3’ hyper-biotinylated antisense oligos corresponding to 4q35
genes will be used as probes for
coding
myotubes.
If
chromatin structure is altered in FSHD, leading to aberrant
then we should not assume that
such a mechanism would affect coding
Non-coding
of both transcription and translation. Although several approaches to the detection of
non-coding RNAs
exist , we propose to use the same technique (
non-coding
(cosmids) will be used to hybridize to these RNAs as the specific non-coding transcripts
cannot be identified a priori.
Lastly,
expression profiling) will be examined in FSHD and control myoblasts/myotubes. A
recent finding in FSHD research within the past year has been the unique and consistent
localization of the 4q telomeric region to the nuclear periphery. While the biological
significance of this localization is not yet known, the existence of nuclear domains either
permissive or repressive of transcription is well documented. Therefore, genes affected in
FSHD will be examined by
FSHD region at the nuclear
periphery might affect
Grant: FSHS-HDY-001
Researcher: Kyoko Yokomori. Ph.D., Associate Professor
Institution:
Department
of
240D Med Sci I
Project Title: “The Molecular characterization of the chromatin structure of the D4Z4 repeat
associated with FSHD."
$30,000
$30,000 6/1/2006 – 5/31/2007 Year 2
$30,000
3/1/2008 – 2/28/2009 Year 3
Goal:
[Provided by applicant]: Facioscapulohumeral muscular dystrophy (FSHD) is an
autosomal dominant hereditary neuromuscular disorder characterized by progressive
degeneration of the upper body muscles. The majority of disease cases is linked to the
deletion of the D4Z4 repeat array in the subtelomeric region of chromosome 4q (4qter).
Since there appears to be no functional open reading frame in this region, it was
hypothesized that the D4Z4 repeat plays a structural role in governing epigenetic
regulation of gene expression critical for proper muscle cell differentiation and functions,
and that the disease is caused by the inability of the shortened D4Z4 to form its
specialized chromatin structure leading to dysregulation of critical gene expression.
However, the exact nature of this chromatin structure, factors required for the regulation,
and the target genes whose dysregulation may directly evoke disease pathogenicity
remain obscure. Therefore, it is vital to understand D4Z4 function in order to address the
etiology and pathogenesis of FSHD.
We found using chromatin crosslinking and immunoprecipitation (ChIP)
analysis that the heterochromatin binding protein HP1, and an essential protein complex
required for chromatid cohesion termed “cohesin”, specifically bind to overlapping
regions within the D4Z4 repeat in human muscle cells. HP1 was shown to associate with
centromeric heterochromatin through interaction with the methylated lysine 9 residue of
histone H3, the hallmark of silenced chromatin, and recruit cohesin to centromeres in S.
pombe and chicken cells. Consistent with this notion, we detected H3K9 methylation in
D4Z4. Intriguingly, both HP1/cohesin binding and H3K9 methylation at this region are
lost in FSHD mutant cells, in which the 4qter D4Z4 is deleted. These results provide the
first direct evidence that 4qter D4Z4 is heterochromatic, and that this special organization
is lost in FSHD. Thus, our results provide further insight into the molecular nature and
pathogenic contribution of this unique repeat sequence in FSHD.
We hypothesize that human HP1 targets cohesin to D4Z4, and together they
mediate proper heterochromatin structure organization required for normal D4Z4
function, which is abrogated in FSHD. To address this, we plan to carry out biochemical
and cytological analyses of the mechanism and function of cohesin and HP1 binding to
D4Z4. Specific aims are 1) analysis of HP1/cohesin binding to D4Z4 in normal and
FSHD cells, 2) characterization of the underlying mechanism and factor requirement for
HP1/cohesin binding to D4Z4, and 3) analysis of the effect of cohesin and HP1 depletion
on chromatin structure organization and function of D4Z4 at 4qter. I believe that the
proposed project will make unique contributions to further understanding of the
chromatin structure of D4Z4 and its role in the development of FSHD and may lead to
possible identification of new therapeutic targets.
FSH Society, Inc. Small Grants
Grant: FSHS-SG-001
Researcher: Valery Kazakov, M.D., Ph.D.
Institution: Department of Neurology, Pavlov's Medical Institute, Lev Tolstoy str. 6/8, 197022, filial
1
Project Title: Travel Grant
$952 4/1997
Goal: To assist with travel to the
AAN/NIH/
Meeting
Grant: FSHS-SG-002
Researcher: Michio Hirano, M.D.
Institution: College of Physicians & Surgeons of Columbia University, P&S 4-443, 630 West 168th
Street,
2nd Floor, Room 401,
Project Title: AAN Travel Fellowship Grant
$1078.87 5/1997
Goal: To assist with travel to the
AAN/NIH/
Meeting
Grant: FSHS-SG-003
Researcher: David Lacomis, M.D.
Institution:
Project Title: AAN Travel Fellowship Grant
$1333.28 5/1997
Goal: To assist with travel to the
AAN/NIH/
Meeting
Grant: FSHS-SG-004
Researcher: William Ted Brown, M.D., Ph.D.
Institution: New
York Institute for Basic Research,
Project Title: Travel Grant
$670.80 5/1997
Goal: To assist with travel to the
Annual
Meeting. For opinion, estimate and consulting on establishing FSHD genetic testing in
the
Grant: FSHS-SG-005
Researcher: Rossella Tupler, M.D., Ph.D.
Institution: Howard Hughes Medical Institute, Program in Gene Function and Expression, University
of
Project Title: Travel Grant
$233 10/1997
Goal: To assist with travel to the
Annual
Meeting as satellite to the ASHG.
Grant: FSHS-SG-006
Researcher: Linda Surh, Ph.D.
Institution: Molecular
Diagnostics Lab, Children's
Room
3029,
Project Title: Equipment Grant
$771.95 12/1997
Goal: To purchase needed equipment for pulse field gel electrophoresis required for accurate
molecular genetics testing of FSHD.
Grant: FSHS-SG-007
Researcher: Meena Upadhyaya, Ph.D.
Institution:
Project Title: Travel Grant
$1,346.14 7/1998
Goal: To assist with travel to the
International Muscle Conference
Grant: FSHS-SG-008
Researcher: Silvere van der Maarel, Ph.D.
Institution:
Project Title: Travel Grant
$845.24 11/1998
Goal: To assist with travel to the
Annual
Meeting as satellite to the ASHG.
Grant: FSHS-SG-009
Researcher: Kevin Flanigan, M.D.
Institution: Eccles
Institute of Genetics, Room 7290,
Project Title: Small grant proposal for “QMA software/system and professional physical therapy
resources to help with studies to answer definitively whether anticipation in disease
severity and onset, gender effects, or parent-of origin effects exist in FSHD.”
$8,375
Goal: To assist with data collection
in clinical trials and patient acquisition at the
effect) in the region.
Grant: FSHS-SG-010
Researcher: Yang (Ted) D. Teng, M.D., Ph.D.
Institution: Children's
Hospital,
Project Title: Travel Grant
$138.52 6/2000
Goal: To assist with travel to the NIH
FSHD Symposium held at
Grant: FSHS-SG-011
Researcher: Amy Csink, Ph.D.
Institution: Department
of Biological Sciences,
Project Title: Travel Grant
$840.70 10/2000
Goal: To assist with travel to the
Annual
Meeting as satellite to the ASHG. Lecture on Apoptosis.
Grant: FSHS-SG-012
Researcher: Laura M. Palmucci, M.D., Ph.D.
Institution: Centro
Malattie Neuromuscolari,
Department of Neuroscience,
Cherasco 15, 10126,
Project Title: Travel Grant
$395.75 10/2000
Goal: To assist with travel to the
Annual
Meeting as satellite to the ASHG.
Grant: FSHS-SG-013
Researcher: Peter Lunt, Ph.D.
Institution: Department
Clinical Genetics,
Michael's
Hill,
Project Title: Travel Grant
$776.93 12/2000
Goal: To assist with travel to the
Annual
Meeting as satellite to the ASHG.
Grant: FSHS-SG-014
Researcher: Michel van Geel, Ph.D.
Institution:
Project Title: Research Publication Grant "Support for doctoral thesis printing efforts."
$2,500 7/2001
Goal: To facilitate printing and dissemination of research doctoral thesis book covering high
level and detailed work in evolutionary aspects of FSHD and including the first notion of
A/B allele in FSHD.
Grant: FSHS-SG-015
Researcher: Patrick Reed, Ph.D.
Institution: Department of
Physiology,
Redwood
St.,
Project Title: Travel Grant
$700 7/2001
Goal: To facilitate travel to the 6th World Muscle Society Congress, Thursday, 6 September
2001, Snowbird, UT to allow fellow to present recent data on FSHD. to facilitate
thinking and research on structural and patho-physiological models for FSHD.
Grant: FSHS-SG-016
Researcher: Rossella Tupler, M.D., Ph.D.
Institution: Howard Hughes Medical Institute, Program in Gene Function and Expression, University
of
Project Title: Travel Grant
$677.82 10/2001
Goal: To assist with travel to the
Annual
Meeting as satellite to the ASHG.
Grant: FSHS-SG-017
Researcher: Rune R. Frants, Ph.D.
Institution: Human
Genetics,
The
Project Title: Travel Grant
$534.23 10/2001
Goal: To assist with travel to the
Annual
Meeting as satellite to the ASHG.
Grant: FSHS-SG-018
Researcher: Richard
Festenstein, Ph.D.
Institution: Gene Control Mechanisms and Disease, Room 5006, Clinical Research Building, MRC
Clinical
Sciences Centre, Department of Medicine,
Road,
Project Title: Travel Grant
$1,500 11/2001
Goal: To facilitate travel to the 6th World Muscle Society Congress, Thursday, 6 September
2001, Snowbird, UT to deliver Keynote FSHD Lecture on: “Chromatin structure, gene
expression, and disease.” To facilitate thinking and research on chromatin structure and
models for FSHD.
Grant: FSHS-SG-019
Researcher: Sara T. Winokur, Ph.D.
Institution: 240 D, Medical
Sciences I, Department of Biological Chemistry,
Project Title: Small Research Grant for “FSHD-Research ListServ.”
$7,000 3//2002 – open
Goal: To facilitate collaboration, sharing and biomaterials collection and distribution in the
FSHD clinical and research communities.
Grant: FSHS-SG-020
Researcher: Davide Gabellini, Ph.D.
Institution: Howard Hughes Medical Institute, Program in Gene Function and Expression, University
of
Project Title: Travel Grant
$508.78 10/2002
Goal: To assist with travel to the
Annual
Meeting as satellite to the ASHG.
Grant: FSHS-SG-021
Researcher: Rossella Tupler, M.D., Ph.D.
Institution: Howard Hughes Medical Institute, Program in Gene Function and Expression, University
of
Project Title: Travel Grant
$678.69 10/2002
Goal: To assist with travel to the
Annual
Meeting as satellite to the ASHG.
Grant: FSHS-SG-022
Researcher: Michael R. Green, M.D., Ph.D.
Institution: Howard Hughes Medical Institute, Program in Molecular Medicine, University of
Project Title: Travel Grant
$720.37 10/2002
Goal: To assist with travel to the
Annual
Meeting as satellite to the ASHG.
Grant: FSHS-SG-023
Researcher: Yukiko K. Hayashi, M.D. and Kanako Goto, Ph.D.
Institution: Department
of Neuromuscular Research, National Institute of Neuroscience,NCNP,
Ogawa-Higashi,
Kodaira,
Project Title:
Grant
$2,761.84 10/2002
Goal: To assist with travel to the
Annual
Meeting as satellite to the ASHG. To facilitate continuity of Japanese FSHD research.
Grant: FSHS-SG-024
Researcher: James Marshall, Ph.D. and Kylie DeBoer, Ph.D.
Institution: Sydney
IVF, Level 11,
Project Title:
Grant
$2,000 10/2003
Goal: To assist with travel to the
Annual
Meeting as satellite to the ASHG. To present initial data on in vitro fertilization
techniques for FSHD to the FSHD research community.
Grant: FSHS-SG-025
Researcher: Valery Kazakov, M.D., Ph.D.
Institution: Department of Neurology, Pavlov's Medical Institute, Lev Tolstoy str. 6/8, 197022, filial
1
Project Title: Research Grant
$650 11/2003
Goal: To assist with cost of
Upadhyaya in
Grant: FSHS-SG-026
Researcher: Kevin Flanigan, M.D.
Institution: Eccles Institute of
Genetics, Room 4420,
Project Title:: Travel Grant
$854.04 11/2003
Goal: To assist
with travel to the Annual
Meeting as satellite to the ASHG.
Grant: FSHS-SG-027
Researcher: Silvere van der Maarel, Ph.D.
Institution:
Project Title: Travel Grant
$956.14 11/2003
Goal: To assist with travel to the
Annual
Meeting as satellite to the ASHG.
Grant: FSHS-SG-028
Researcher: H. Lee Sweeney
Institution:
Project Title: Conference Grant
$500 12/2003
Goal: Conference Support Grant for 1st "New Directions in Biology and Disease of Skeletal
Muscle,"
Grant: FSHS-SG-029
Researcher: Alberto Luis Rosa, M.D., Ph.D.
Institution:
Project Title:
Grant
$1,506.02 7/2004
Goal: To assist with travel to the 7th Annual Summer School in Myology to lecture on FSHD
with
J. Adoni Urtizberea, M.D.
and for travel to
with Alexandra Belayew, Ph.D. on DUX4 and FSHD.
Grant: FSHS-SG-030
Researcher: Silvere van der Maarel, Ph.D.
Institution:
Project Title: Travel Grant
$1,184.97 11/2004
Goal: To assist with travel to the
Annual
Meeting as satellite to the ASHG.
Grant: FSHS-SG-031
Researcher: George W.A.M. Padberg, M.D., Ph.D.
Institution: c/o Anjali Kali,
Department of Neurology, 326,
HB
Nijmegen, The
Project Title: Travel Grant
$1,656.19 12/2004
Goal: To assist with travel to the
Annual
Meeting as satellite to the ASHG.
Grant: FSHS-SG-032
Researcher: Richard Lemmers, Ph.D.
Institution:
Project Title: Research Publication Grant
$851.19 03/2005
Goal: To assist with publication, production and distribution of doctoral thesis on FSHD.
Grant: FSHS-SG-033
Researcher:
Institution:
Endocrinologie C4-R, Albinusdreef
2, 2333 ZA,
Project Title: Research Publication Grant
$851.19 03/2005
Goal: To assist with publication, production and distribution of doctoral thesis on FSHD.
Grant: FSHS-SG-034
Researcher: Silvana van Koningsbruggen, Ph.D.
Institution:
Project Title: Research Publication Grant
$851.19 03/2005
Goal: To assist with publication, production and distribution of doctoral thesis on FSHD.
Grant: FSHS-SG-035
Researcher: Kristen Bastress and Marcy Speer, Ph.D.
Institution:
Project Title: Research Project Grant
$3,800 04/2005
Goal: To assist with travel to
FSHD samples with distal and proximal deletions in May 2005.
Grant: FSHS-SG-036
Researcher: Meena Upadhyaya, Ph.D.
Institution:
Project Title: Travel Grant
$1,383.15 10/2006
Goal: To assist with travel to the
Annual
Meeting as satellite to the ASHG.
Grant: FSHS-SG-037
Researcher: Elly
L. van der Kooi, M.D.
Institution: Neuromuscular
Centre
Project Title: Research Publication Grant
$2,448.04 12/2006
Goal: To assist with publication, production and distribution of doctoral thesis on FSHD.
Titled: “Facioscapulohumeral Muscular Dystrophy: interventions and intervention
studies.”
Grant: FSHS-FSHS-SG2007
Researcher: Silvere van der Maarel, Ph.D.
Institution:
Project Title: Small Grant for Support of Prof. Dr. Silvere van der Maarel’s
“Inaugural
Lecture at
visibility for facioscapulohumeral muscular dystrophy (FSHD) and
Epigenetics”
Grant: FSHS-FSHS-SG2007
Researcher(s): Alberto L. Rosa, M.D., Ph.D & E. Daniel Corona
Institution: INIMEC - CONICET
Project Title: “2007 Kiichi Arahata, M.D. Memorial Travel Grant” for ”2007
FSH Society FSHD International Research Consortium workshop
at the American Society of Human Genetics in
Grant: FSHS-FSHS-SG01-2008
Researcher: Alexandra
Belayew, Ph.D.
Institution:
University
of Mons-Hainaut
Project Title: “For Services of Production of 30-50 mg of the 9A12 antibody @ 30 €/mg for DUX4/DUX4c”
Goal:
[Provided by applicant]: Funds for the services of production of 30 - 50 mg of the
9A12 antibody @ 30 €/mg.
List of researchers who have received our 9A12 monoclonal
antibody raised against DUX4/DUX4c:
Yi Wen
Dalila Laoudj-Chenivesse,
Michael Kyba,
Frédérique
Magdinier,
Peter Ambros,
Sara Winokur,
Silvere Van der Maarel, Leiden, The Netherlands
Davide Gabellini,
Stephen Tapscott,
Brian Kennedy,
Patrick Reed,
Meredith Hanel,
Harper Scott,
Luis Garcia,
Total: $