Funded Research
Funded Research

A Xenograft Model of FSHD for Testing Therapeutics

Budget

• Amount awarded: US$100,000
• FSHD Canada Foundation Contribution: US$20,000
• Date awarded: June 20, 2017
• End date: June 20, 2019

Lay Abstract

Investigator: Robert J Bloch PhD, Professor of Physiology
 at the University of Maryland

Facioscapulohumeral muscular dystrophy (FSHD) affects 1 in approximately 15,000 individuals worldwide. One of the obstacles in developing treatments for FSHD is the lack of a small animal model of the disease. The genetics of FSHD make it difficult, if not impossible, to make a mouse with muscles that have the same characteristics as the muscles of individuals with FSHD. This in turn has made it difficult to study mature muscle fibers with the characteristics of FSHD muscle in the laboratory, where conditions can be controlled to examine the morphological and physiological changes associated with disease and to test therapeutics that can reverse those changes. We have taken a novel approach to creating a mouse with FSHD muscles: we are transplanting human muscle precursor cells (hMPCs) from healthy and FSHD donors into mice and inducing them to form mature muscle tissue. We pretreat the mice to eliminate the major hindlimb muscle, the Tibialis anterior, and to prevent it from regenerating. We then inject hMPCs and treat the muscle with an unique set of methods to promote their survival and their ability to form new muscle of human origin. We have had great success using both normal and FSHD MPCs to form human muscle fibers in the mouse hind limb that are mature and functional. Moreover, the muscles formed from FSHD cells express DUX4, the gene that is altered in FSHD and that is most likely to cause the disease.

Here we propose to collaborate with Fulcrum Therapeutics to generate a genetically well controlled set of grafts from cells from a mosaic patient with FSHD. The grafts will be derived from cells that carry the genetic defect associated with FSHD, or from normal cells from the same individual. We will characterize these grafts and then use them to test a new therapeutic drug developed by Fulcrum to target DUX4.

Developing LNA-based therapy for FSHD

Budget

• Amount Awarded: $179,104 for two years
• FSHD Canada Foundation Contribution: US$20,000
• Date Awarded:04/01/2017
• End Date: 03/31/2019

Lay Abstract

Investigators:
• Yi-Wen Chen Children’s National Health System (Washington DC)
• Toshifumi Yokota University of Alberta Faculty of Medicine and Dentistry (Alberta, Canada)

Yi-Wen Chen and Toshifumi Yokota are investigating one of the most promising antisense oligonucleotides (AON) compounds called LNA (locked nucleic acid) gapmer for its efficacy in reducing DUX4 in cell culture and in a mouse model of FSHD. AONs are short gene-like molecules that bind to and inactivate target genes (in this case DUX4). LNA gapmers are a “third generation” AON designed to overcome some problems that made earlier AONs unsuitable for use as therapeutics. LNA gapmers are more stable, resistant to being degraded, and can penetrate the cell membrane and get into cell nuclei where the target DUX4 gene resides. Dr. Yokota will continue to improve the anti-DUX4 LNA gapmer, testing them in FSHD cell lines, while Dr. Chen will test the safety and efficacy of the molecules in a mouse model of FSHD.

Facio Therapies

Budget

FSHD Canada Foundation made a C$100,000 investment in Facio Therapies in late 2016.

Lay Abstract

Facio Therapies (Facio-therapies.com) is a company based in the Netherlands that is focused on finding a cure for FSHD. Facio have screened over 34,000 small molecule compounds, and obtained over 300 so-called ‘hits’ and identified novel approaches to repress the production of the muscle-toxic DUX4 protein. Facio are now preparing for the development of lead compounds for preclinical and clinical development.

FSHD myofibers cultured from human xenografts

Budget

$100,000 (USD) in total, supported in partnership with Friends of FSH Research (50:50)

Lay Abstract

FSH Muscular Dystrophy (FSHD) affects 1 in approximately 8,300 individuals worldwide. One of the obstacles in developing treatments for FSHD is the lack of a small animal model of the disease. The genetics of FSHD make it difficult, if not impossible, to make a mouse with muscles that have the same characteristics as the muscles of individuals with FSHD. This in turn has made it difficult to study mature muscle fibers with the characteristics of FSHD muscle in the laboratory, where conditions can be controlled to examine the morphological and physiological changes associated with disease and to test therapeutics that can reverse those changes.

We have taken a novel approach to creating a mouse with FSHD muscles: we are transplanting muscle precursor cells (MPCs) from healthy and FSHD donors into mice and inducing them to form mature muscle tissue. We pretreat the mice to eliminate the major hindlimb muscle, the Tibialis anterior, and to prevent it from regenerating. We then inject MPCs and treat the muscle with an unique set of methods to promote their survival and their ability to form new muscle of human origin. We have had great success using both normal and FSHD MPCs to form human muscle fibers in the mouse hindlimb that are mature and functional. Moreover, the muscles formed from FSHD cells express DUX4, a marker of the disease.

We have recently succeeded in isolating muscle fibers from the muscle grafts and have begun to study them in culture dishes. Here we propose to develop these methods and to use them to study DUX4 in isolated FSHD muscle fibers to learn how DUX4 might lead to FSHD.

Determining the effectiveness of increased SMCHD1 expression to suppress DUX4 in FSHD muscle cells and model mice

Budget

$50,566 (USD) in total, supported in partnership with the FSH Society (50:50)

Lay Abstract

There are two genetic types of FSHD: FSHD1 and FSHD2. FSHD1 (representing around 95% of FSHD cases) is associated with a loss of repetitive DNA units called D4Z4 on the end of chromosome 4.  The remaining 5% of FSHD cases are FSHD2, and the majority of these cases are associated with mutations in a gene known as SMCHD1, located on chromosome 18. This gene helps to maintain the repetitive D4Z4 region associated with FSHD1. Further, it is also known that SMCHD1 modifies the severity of symptoms in patients with FSHD1.

In both FSHD1 and FSHD2, these genetic changes result in the activation (or expression) of a gene called DUX4, which is toxic to muscle. This gene is normally not active in adult muscles, but in FSHD DUX4 gets expressed and is thought to weaken and kill muscles. In this project, Dr. Hiramuki will manipulate the level of SMCHD1 and study how this affects the expression of DUX4, comparing cells with 7-10 D4Z4 repeats (corresponding to less severe FSHD) and 1-6 repeats (more severe FSHD). Similarly, he will investigate different versions of SMCHD1 for their effectiveness in suppressing DUX4. These are proof of principle experiments showing that higher SMCHD1 will be effective as a potential therapy.

In a second set of experiments, Dr. Hiramuki will use a genetically engineered virus (called adeno associated virus 6, or AAV6) to introduce extra SMCHD1 into human FSHD muscle cells, as well as in mice. The hope is that this will increase levels of SMCHD1 and suppress DUX4. With these experiments, he hopes to develop a gene therapy method that one day could be used to deliver SMCHD1 into FSHD patients’ muscles.

FSHD myofibers cultured from human xenografts

Budget

$100,000 (USD) in total, supported in partnership with Friends of FSH Research (50:50)

Lay Abstract

Much of what we know about DUX4 and its associated toxicity comes from cultured cells from FSHD patients. We basically take cells from a biopsy, grow them on a dish, and learn something about DUX4. DUX4 is a pretty toxic molecule. Most cell types that you introduce it to are horrified by its presence and they undergo cell death. However, there are major discrepancies, and a serious lack of consensus for how DUX4 actually kills muscle in a patient with FSHD. Part of this stems from the fact that DUX4 in a patient is incredibly difficult to find. In fact, in most cell cultures of FSHD muscle, DUX4 is not very easy to detect. How could such a little bit of a bad protein cause such big problems? Whereas it’s easy to imagine how a lot of the protein can cause problems – we know that it does – it’s more difficult to understand how trickles or bursts of the protein cause irreparable muscle damage.

Dr. Bloch has been developing an animal model of FSHD using myoblasts from patients. Here, he takes these FSHD muscle precursor cells (that have been modified so that they divide rapidly) and he implants them into the muscle of a mouse that has been damaged by radiation and chemicals.

The engraftment he sees is amazing. These mice have muscle that is mostly made up of human cells. This is obviously a model we’ll want to use for drug discovery at some point, but for now Dr. Bloch is proposing to do something a little bit out of the box.

Dr. Bloch proposed to isolate complete human myofibers from the mice. In mice, the ability to isolate a single muscle fiber from an intact muscle has taught us a great deal about the biology of skeletal muscle. There are some fundamental questions we can ask here. What rate is DUX4 activated in the muscle fibers? What percentage of muscle fibers make DUX4? What defects arise? The studies may ultimately inform how we pursue the analysis of DUX4 and its associated toxicity in human. This will then directly inform how we go about treating the disease.

Investigation DUX4 activity during development of hESC-derived skeletal muscle.

Budget

$50,000 (USD) in total, supported in partnership with Friends of FSH Research (50:50)

Lay Abstract

In 2011, FSHD Global funded Genea Biocells to isolate the first embryonic stem cells from patients with FSHD. Genea has done an excellent job developing these lines, and sharing them with the research community. Here we are leveraging Friends of FSH Research's expertise in the development of assays for high throughput screening to support the creation of an engineered embryonic stem cell line that reports DUX4 activity.

This tool will be able to be used by researchers all over the world to perform basic research and screen for lead compounds that interfere with DUX4 expression. We have funded a number of screens previously, and these efforts have resulted in the identification of lead compounds.

There are only two ways to develop pharmacological drugs for things that reduce or inhibit DUX4. The first is to screen libraries of drugs, and the second is to design. To design you need to know the structure of DUX4, and we’re working on that (see Hideki Aihara’s grant we funded last cycle). In the meantime, we’ll screen libraries.

Screening a drug library is only as good as the assay that is being used. We have had success using a number of approaches: researchers have put DUX4 in to cancer cells for example, and looked for things that shut the artificial DUX4 down. But the thing is, if you’re artificially introducing DUX4 into cells, then you won’t detect drugs that may in fact prevent DUX4 from turning on in the first place.

If you want to do that, you have to do the experiment on a human cell from an FSHD patient. We’ve invested in those types of studies before, and those also generated potential leads. The problem with relying on human cells (in particular muscle cells) for screening, is they are difficult to grow, unpredictable, and tough to scale if you want to screen a lot of drugs (and believe me there are a LOT of drugs to screen in this world, ranging from libraries of FDA approved drugs to structure based libraries that look for classes of compounds).

RNA-mediated Epigenetic Silencing of D4Z4 repeats: Implications for targeted therapy for FSHD

Budget

$60,180 (USD) in total, supported in partnership with Friends of FSH Research (50:50)

Lay Abstract

Dr. Jong Won Lim is a Friends of FSH Research Fellow at the Fred Hutchinson Cancer Research Center in Seattle. He is mentored by Dr. Stephen Tapscott and Dr. Galina Filippova, and has recently published the results from his first round of funding from Friends of FSH Research.

Dr. Lim and team have been resilient in developing the experiments needed to help answer one of the most fundamental questions in FSHD biology - and at the same time have identified a potential way to intervene with disease progression. Whereas some projects we fund are easy to explain to our supporters, this one is not, so bear with me and follow along!! As always, we are here to answer any questions you may have.

The fundamental question underlying this work is why does DUX4 turn on when there are less than 10 copies in a row? (if what I just wrote made no sense, take a minute to review the mechanism of FSHD). We've suspected for a long time that the reason is that the DUX4-containing array itself emits some kind of signal telling the cell to shut it down. But what on earth could those signals be? It's not like a piece of DNA has a flare gun it can light - does it?

Before we delve too deep, let's talk therapeutics. DUX4 is a protein that is not supposed to be turned on. Because of the nature of the protein, and the fact that it is specifically expressed in patients with FSHD, the most logical strategy for therapy is to stop it from working. This may be a traditional pharmaceutical drug, and we are actively investigating whether there are drugs that will do the job (here is one example). However, there are also experimental therapeutics that target specific DNA sequences using DNA or RNA sequences that match the DUX4 genetic sequence. How's that for futuristic! Recent progress with treatment of spinal muscular atrophy highlights the potential of this technology, and hopefully we will one day be able to write an article like this one.

DNA drugs are a particularly logical approach for FSHD - in fact, we've recently funded a study to test the efficacy of such a compound (soon to be announced) , and yet another that seeks a novel way of targeting the DUX4 gene. Both of these compounds work by interfering with the DUX4 mRNA. If you mess up the DUX4 mRNA, DUX4 protein can't be made. But the mRNA is still made because you haven't solved the root of the problem: the DNA encoding DUX4 is not silenced. As a creative analogy, it's like stopping a baby from crying by putting in earplugs.

In a nutshell, Dr. Lim's study shows that small chemically modified RNA molecules that target the sequences surrounding the DUX4 gene are capable of re-silencing the DUX4 array. He has swaddled the baby and overfed it.

Here's what we think is happening. DUX4 is a gene that is encoded in a head to tail array - think of it like a conga-line dancing to Hot Hot Hot in your cells. In between each gene is a sequence that governs the gene's expression. Think of that sequence as the hands in the conga line. Usually, that regulatory sequence pushes gene expression in one direction so only one gene is made (i.e. your hands in the conga line only touch the hips of the person in front of you) - but we already know this is not the case for the DUX4 array.

We know that the regulatory sequence within the array is bidirectional, meaning it can drive expression of gene expression in both directions. One hand is grabbing the hips of the person in front, and the other is grabbing another body part of the person behind. This means you are generating two different RNAs, and because they are opposite from one another, they can bind one another. When two RNA molecules come together that is the equivalent of firing a flare gun off in the cell.

These double stranded molecules feed into a cellular defense mechanism so that the invading RNA is rapidly destroyed. One of the proteins involved in that defense mechanism is called Argonaut (AGO). To make a long story short, Dr. Lim showed that the DUX4 array is shedding off small pieces of RNA that are getting loaded onto an AGO protein, and finding their way back to the array and telling the cell to shut it down. He also showed that proteins required for the generation of siRNA are necessary for the effect.

Because of the superb progress made on this project, Friends of FSH Research has renewed Dr. Lim's funding for an additional year.

Validation of a PPMO morpholino lead compound in an animal model of FSHD

Budget

$50,000 (USD) in total, supported in partnership with Friends of FSH Research (50:50)

Lay Abstract

In a very nice review recently published in “Trends in Molecular Medicine”, Lek et al. discussed the over 10 animal models that have been developed for FSHD. That’s quite a different story than a few years ago, when there were no animal models. The purpose of these animal models is to test the efficacy of drugs that may be potentially therapeutic for FSHD. Dr. Gregory Block has written before how the nature of the drug/compound that is being tested will likely reflect which animal model we choose to use.

For example, if the lead compound targets DUX4 directly, it will suffice to simply introduce the DUX4 gene into an animal. That is what Dr. Harper has done at Nationwide Children’s Hospital in Columbus Ohio. Leveraging his expertise in adeno-associated virus (AAV) delivery of genes to skeletal muscle, Dr. Harper and team have shown that introduction of DUX4 into the muscle of mice results in the destruction of the injected muscle.

Genzyme, a biotechnology company that focuses on therapeutics for rare diseases, has developed an antisense oligonucleotide-based approach for targeting the DUX4 transcript for degradation. Make no mistake, this is not a novel approach. There are multiple projects running throughout the world looking to develop such molecules, and they are very easy to get working in cell models of disease. Where Genzyme has an advantage is in the chemistry to deliver these molecules to skeletal muscle in an animal or human. They have considerable expertise in advancing these therapeutics rapidly.

This project is one of the first that aims to test a lead compound in one of the animal models that has been developed by the community, and thus is a major milestone in the field. This model was chosen because of its simplicity. The drug can be delivered systemically, and the efficacy can be measured against a variety of concentrations of DUX4. Thus we can create a full pharmacological profile for the lead compound and understand its efficacy and toxicity in the animal model.

If this study is successful, it will pave the path for the group to aim towards larger animal studies that will generate the data needed to justify a clinical trial.

This study was co-funded by FSHD Canada.

Classification of DUX4 expression in human myofibers derived from xenogeneic muscle cell transplants.

Budget

$50,000 (USD) in total, supported in partnership with Friends of FSH Research (50:50)

Lay Abstract

Much of what we know about DUX4 and its associated toxicity comes from cultured cells from FSHD patients. We basically take cells from a biopsy, grow them on a dish, and learn something about DUX4. DUX4 is a pretty toxic molecule. Most cell types that you introduce it to are horrified by its presence and they undergo cell death. However, there are major discrepancies, and a serious lack of consensus for how DUX4 actually kills muscle in a patient with FSHD. Part of this stems from the fact that DUX4 in a patient is incredibly difficult to find. In fact, in most cell cultures of FSHD muscle, DUX4 is not very easy to detect. How could such a little bit of a bad protein cause such big problems? Whereas it’s easy to imagine how a lot of the protein can cause problems – we know that it does – it’s more difficult to understand how trickles or bursts of the protein cause irreparable muscle damage.

Dr. Robert Bloch has been developing an animal model of FSHD using myoblasts from patients. Here, he takes these FSHD muscle precursor cells (that have been modified so that they divide rapidly) and he implants them into the muscle of a mouse that has been damaged by radiation and chemicals.

The engraftment he sees is amazing. These mice have muscle that is mostly made up of human cells. This is obviously a model we’ll want to use for drug discovery at some point, but for now Dr. Bloch is proposing to do something a little bit out of the box.

Dr. Bloch proposed to isolate complete human myofibers from the mice. In mice, the ability to isolate a single muscle fiber from an intact muscle has taught us a great deal about the biology of skeletal muscle. There are some fundamental questions we can ask here. What rate is DUX4 activated in the muscle fibers? What percentage of muscle fibers make DUX4? What defects arise? The studies may ultimately inform how we pursue the analysis of DUX4 and its associated toxicity in human. This will then directly inform how we go about treating the disease.

Exploiting Genome Editing Technology 2015

Budget

$125,000 in total, supported in partnership with the FSH Society (50:50)

Lay Abstract

FSHD Canada and the FSH Society are jointly funding a project called "Exploiting Genome Editing Technologies," led by Michael Kyba, PhD, of the University of Minnesota. The two FSHD organizations joined forces last year to support the first year of the US$125,000 project, with each putting in US$62,500. Based on the success of the first year, they agreed to fund the second year of the project for the same amount.

"The past several years have seen tremendous advances in our ability to specifically modify DNA sequences in the human genome," said Kyba. "FSHD is caused by the combination of two factors: the contraction of D4Z4 repeat number combined with the presence of a pathogenic sequence downstream of the D4Z4 repeats. Both of these DNA elements are accessible to modification using newly developed genome editing technology. The Kyba lab has developed methods of gene targeting that address both elements and will use these to derive and study genetically corrected FSHD cell lines."

FSHD Canada and Friends of FSH Research Team Up to Fund Three Innovative Research Projects.

Budget

FSHD Canada funded half of each project. Dr. Parker: $50,000. Dr. Wagner/Natoya: $50,000. Dr. Aihara/Dr. Kyba: $33397.

Lay Abstract

FSHD Canada has partnered with Friends of FSH Research to solicit, review, and fund three exciting projects that will accelerate our progress towards a treatment or cure for FSH muscular dystrophy.

This cycle, we received more applications than ever for projects and choosing the final three was quite difficult. The first project that was accepted by the Friends’ Scientific Advisory Board was from Dr. Eric Wagner at the University of Texas. Dr. Wagner and his graduate student, Natoya Peart, have done some amazing work understanding how RNA molecules get packaged. By specifically focusing on sequences within DUX4, the candidate target gene for FSHD, they believe they can develop novel DNA therapeutics. DNA therapeutics are not entirely a pipe-dream. There are a number that are being developed for clinical trials in other diseases. According to Dr. Wagner, these therapeutics are gaining a “resurgence in clinical trials” and some have been given approval by the FDA.

The second project being co-funded is with Drs. Hideki Aihara and Michael Kyba. Their project focuses on two elements of the structure of DUX4. First, it will identify the optimal binding characteristics of DNA bound by DUX4, and second, will seek to identify the structure of the DUX4 protein bound to that DNA. Understanding the structure of DNA and how it interacts with it’s target DNA is critical for the development and fine tuning of compounds designed to interfere with DUX4 function.

Last, but certainly not least, based on a successful review or her 1 year project that was co-funded last year, FSHD Canada and Friends of FSH Research have continued funding Dr. Maura Parker, as she continues to develop the methodologies for isolating and classifying a new class of muscle precursor cells that will function to repopulate the skeletal muscle of mice, as well as to improve cellular models of FSHD.

Friends of FSH Research is a Seattle based non-profit that raises money, solicits, reviews, and funds research towards a treatment or cure for FSH Muscular Dystrophy.

Links of Interest: Friends of FSH Research

FSHD Canada Co-Funds Two Very Promising Research Projects in Collaboration with the Friends of FSH Research

Budget

FSHD Canada funded half of each project. Dr. Miller/Tawil: $44,498. Dr. Kyba/Tavassoli: $25,000.

Lay Abstract

We are excited to announce that FSHD Canada has co-­funded two very promising research projects in collaboration with the Friends of FSH Research.

Donations like yours have contributed towards accelerating our progress towards understanding FSH. Knowledge truly is hope, and because of the studies we’ve been able to fund, we now have a better understanding of how to steer the research towards a cure.

There is now a consensus in the research community that a protein called DUX4 is playing a central role in the initiation of muscle deterioration. Finding a mechanism or strategy to inhibit DUX4 from doing damage to muscle is a critical step towards developing treatments. We have funded Dr. Michael Kyba at the University of Minnesota to conduct a massive screening experiment looking for novel inhibitors of DUX4. Using a novel platform developed by Dr. Ali Tavassoli at the University of Southampton, Dr. Kyba will be able to test tens of millions of potential small molecules for the ability to inhibit DUX4 activity. Dr. Kyba has an excellent track record performing large scale screens, and recently has published his first results from the first DUX4 inhibitor screening effort.

The second project we have funded was proposed by Dr. Daniel Miller and Dr. Rabi Tawil at the University of Washington and University of Rochester respectively. Drs. Miller and Tawil have collected serum samples from a group of patients who volunteered to donate blood samples to the laboratories. They will test the blood using a novel technique to search for serum biomarkers that are specific for FSH. Much can be learned through such a study. Not only might Drs. Miller and Tawil identify new and robust markers that can be used in later stage clinical trials, but by testing a wide array of serum biomarkers we may gain critical insight into the mechanisms of muscle damage.

These studies not only fuel our hope that one day we will have a treatment for FSHD, but also represent how very far we have progressed along the path to that cure. We look forward to keeping you up-­to-­date on the status of the current projects.

Links of Interest: Friends of FSH Research, Kyba Website, Miller Website, Tavassoli Website, Tawil Website

Exploiting genome editing technology to modify and regulate the FSHD disease locus

Budget

$125,000 in total, supported in partnership with the FSH Society (50:50)

Michael Kyba, Ph.D. – Lillehei Heart Institute, University of Minnesota (Minneapolis, MN)

Lay Abstract

Recent discoveries of DNA-binding factors have opened up tremendous new possibilities in genome editing. Through the grant, this study will take advantage of and leverage an existing research program in genome editing of FSHD iPS cells, and will provide the field with valuable new tools to study the pathogenesis of FSHD, and to develop cell therapies based on corrected, isogenic, iPS cells.

Generating FSHD-affected human muscle in mice for testing therapeutic strategies

FSHD Canada and Friends of FSH Research are pleased to support Dr. Maura Parker’s project entitled "Generating FSHD-affected human muscle in mice for testing therapeutic strategies." This proposed project aims to generate human FSHD-affected muscle within mice. These mice will be important for studying the disease in the context of an individual, yet sparing patients the pain of multiple muscle biopsies. Most importantly, these mice will be essential for pre-clinical testing of potential therapies.

This is the third in a series of grants funded by a partnership of FSHD Canada and Friends of FSH Research, allowing your generous donations to go even farther.

Budget

$39,929.65 for 1 year

Lay Abstract

Great advances have been made to understand the cause of FSHD, and the mechanism behind the disease. Currently, researchers indicate that improper expression of DUX4 in FSHD-affected muscle is the cause of the disease. However, to further understand FSHD and test potential therapies, we need to develop an appropriate pre-clinical animal model. Mice are the most widely used animal model, as they are easy to handle and breed. However, mice do not have the DUX4 gene in their genome, suggesting that expression of DUX4 protein within mouse muscle may not fully reproduce the human disease. Therefore, we propose to generate human FSHD-affected muscle within mice. We will do this by transplanting muscle stem cells from FSHD-affected individuals into the regenerating muscle of mice. Based on our current studies, we expect to generate a muscle within the mouse that is 90% human-derived. These mice will be important for studying the disease in the context of an individual, yet sparing patients the pain of multiple muscle biopsies. Most importantly, these mice will be essential for pre-clinical testing of potential therapies.

Identification of Chromatin Modifier Genes Important for DUX4 Silencing

FSHD Canada and Friends of FSH Research are pleased to support the Daniel G. Miller MD PhD & Galina Filippova PhD project entitled "Identification of Chromatin Modifier Genes Important for DUX4 Silencing."

Budget

$50,000 for 1 year

Lay Abstract

Facioscapulohumeral Muscular Dystrophy is a dominantly inherited myopathy associated with chromatin relaxation of the D4Z4 macrosatellite array on chromosome 4. DUX4 is encoded within each unit of the D4Z4 array where it is normally transcriptionally silenced and packaged as constitutive heterochromatin. Truncation of the array to less than 11 D4Z4 units (FSHD1) or mutations in SMCHD1 (FSHD2) results in chromatin relaxation and a small percentage of cultured myoblasts from these individuals exhibit infrequent bursts of DUX4 expression. This proposal builds on the hypothesis that epigenetic modification of D4Z4 chromatin structure is an important strategy for disease treatment. Here we propose to construct a library of siRNA’s that target genes involved in chromatin modification and examine the effect of reducing transcript levels on DUX4 gene expression to determine integral components of the DUX4 transcriptional silencing machinery.

Identification of small RNAs generated from the D4Z4 Array

FSHD Canada and Friends of FSH Research are pleased to support the Galina Filippova PhD & Daniel G. Miller MD PhD project entitled "Identification of small RNAs generated from the D4Z4 Array".

Budget

$30,000 for 1 year

Lay Abstract

Facioscapulohumeral muscular dystrophy (FSHD) is caused by incomplete repression of the D4Z4 macrosatellite repeat array on the disease-permissive chromosome 4q that results in aberrant expression of DUX4, the candidate FSHD gene imbedded within the D4Z4 repeat. Previously, we have shown that multiple sense and antisense transcripts as well as small RNAs are expressed from D4Z4 units. We hypothesize that small RNAs generated from D4Z4 long non-coding sense and/or antisense transcripts are involved in heterochromatin formation by recruiting chromatin modifiers to the D4Z4 target regions and leading to increased repressive chromatin modifications and DNA methylation that result in DUX4 epigenetic silencing. Here we propose to identify these RNAs by deep sequencing of small RNA populations in cells from FSHD-affected and unaffected people.

Derivation of human induced pluripotent stem cells from FSH patient fibroblasts

The FSHD Canada Foundation and the FSH Society of the United States announce first joint effort to fund technology that could lead to new insight into the FSHD disease process while providing critically important tools for developing new therapies.

Dr. Gabsang Lee (Johns Hopkins University, Baltimore, Maryland, USA) (2013-2014)

Budget

$49,705 over 1 year

Lay Abstract

The genetic and biological events that result in Facioscapulohumeral muscular dystrophy (FSHD) pathogenesis are complex and the link between the genetic aberration and manifestation of symptoms is still elusive. We hypothesize that there might be cellular and genetic alterations in the early stage of myogenesis in FSHD patients. The establishment of human induced pluripotent stem cells (hiPSCs) ushered in a new era in biomedicine and can be useful for modeling pathogenesis of human genetic diseases, autologous cell therapy after gene correction, and personalized drug screening. Our lab has been studying human genetic disorders by using induced pluripotent stem cells (hiPSCs), a new type of stem cell generated without destruction of any embryonic tissues or embryos. In addition, we already built a novel methodology in a highly innovative manner to directly derive and prospectively isolate skeletal muscle from the hiPSCs. Here we propose to establish hiPSC lines from FSHD patients' somatic cells. Our proposed study will enable us to isolate FSHD-specific skeletal muscle cells for better understanding of FSHD pathogenesis in the human system as well as for advancing potential autologous cellular therapy to correct genetic defects in FSHD in the near future.

https://twitter.com/#!/FSHDCanada/ http://www.facebook.com/pages/FSHD-Canada/331378503625267 /contact Suite 201, 1100 1st St. SE Calgary, Alberta T2G 1B1 403.470.0141 neil.camarta@fshd.ca