MDA awarded a research grant totaling $330,000 to Stanley H. Appel, chair of the department of neurology at the Methodist Neurological Institute (MNI) in Houston, to study the protective effects of a specific class of immune system cells in ALS (amyotrophic lateral sclerosis, or Lou Gehrig's disease).

Appel, a longtime MDA adviser and director of the MDA/ALS center at MNI, will look at T cells, observed in people with ALS as well as in mouse models of the disease, and known to be involved in protecting motor neurons (nerve cells that activate muscles).

Appel's lab previously has documented that T cells can protect neurons in a mouse model of ALS, at least in part by enhancing the neuroprotective functions of another type of immune system cell called microglia. When activated, these cells can produce helpful proteins, but they also can cause dangerous inflammation.

Finding ways to protect motor neurons is a key goal in the development of ALS therapeutics, and Appel's group is working to understand the interaction among other cells in the immune system responsible for prompting T cells to protect neurons.

Project plans include the transplantation of various T-cell types into ALS research mouse models in order to determine which of the cells are most helpful to neurons. A greater understanding of this T-cell population and its associated molecular signals may lead to therapeutics based on increasing the numbers and effectiveness of these cells to help protect neurons in people with ALS.

"MDA has been a critical source of support throughout the development of our immunological studies in ALS," Appel said. "It's permitted us to assemble a team of scientists focused on the neuroinflammatory processes in ALS, and to substantiate the importance of protective immunomodulation in ALS."

Funding for this MDA grant began August 1, 2010.

MDA awarded $352,188 to Zolt Arany, assistant professor in medicine at Beth Israel Deaconess Medical Center, part of Harvard Medical School in Boston, for research into the role of skeletal muscle metabolism in Duchenne muscular dystrophy (DMD).

"It has become increasingly clear that skeletal muscle metabolism plays a critical role both in the onset of, and resistance to, neuromuscular diseases like DMD," Arany said. "Blood vessels are a critical component of metabolism, because they bring oxygen and nutrients to metabolically active tissues."

Arany's team recently uncovered a metabolic pathway that strongly induces the formation of new blood vessels in muscle. Previous work by others already has established that the pathway protects against muscle damage in a model of DMD, but it's not understood how it accomplishes this.

The new work is designed to test the hypothesis, in the mdx mouse model of DMD, that the metabolic pathway protects against muscle degeneration and atrophy by boosting the density and activity of blood vessels.

"Without support from the MDA, we would not be able to initiate these important studies," Arany said. "MDA has had an important impact on research in muscle dystrophy, and its role now, in times of federal budgetary constraints, is more critical and influential than ever."

A greater understanding of precisely how this particular metabolic pathway protects skeletal muscle may lead to novel therapeutic approaches for DMD and other devastating muscle diseases.

Funding for this MDA grant began August 1, 2010.

MDA awarded a grant totaling $303,438 to Kurt Beam, professor in the department of physiology and biophysics at the University of Colorado School of Medicine in Denver, for research into a process called excitation-contraction coupling responsible for the contraction of muscle cells necessary for voluntary movement and breathing.

"Excitation-contraction coupling is a fundamental process in skeletal muscle that is not yet well-understood," Beam said. "Additionally, mutations of the key proteins involved in excitation-contraction coupling cause serious human muscle diseases, including periodic paralyses, malignant hyperthermia and central core disease."

The process is initiated by electrical impulses which move along the membrane that separates the inside and outside of each muscle cell, and depends, at least in part, on two proteins called DHPR and RyR1.

In completed studies, Beam and his group have shown that mutations of either DHPR or RyR1 affect the behavior of both proteins. In their new work, the investigators will study cultured mouse muscle cells as they attempt to uncover the molecular mechanism for communication between DHPR and RYR1.

"The importance of MDA to neuromuscular research cannot be measured in dollars alone. In particular, MDA has been at the forefront in promoting basic and applied research directed toward understanding normal muscle function, mechanisms of muscle disease and development of therapies for these diseases," Beam said. "An especially important role that MDA plays is to provide funding during early career stages and at times when funding from other sources becomes scarce. I do not believe that I would have been able to establish and maintain my neuromuscular research had it not been for funding from MDA at crucial points in my career."

Funding for this MDA grant began August 1, 2010.

Michael Benatar, associate professor of neurology and epidemiology at Emory University in Atlanta, received an MDA grant totaling $525,000 to continue research into the early stage of FALS — familial, or inherited, ALS (amyotrophic lateral sclerosis, or Lou Gehrig's disease) — prior to symptom onset.

With funding from MDA, Benatar began his "pre-FALS" study in 2007, tracking a group of 30 people at risk for developing ALS because they harbor a mutation in the SOD1 gene, known to cause some forms of the disease. His team collected a series of physical, functional and neurological data over time in an effort to discern early biological markers ("biomarkers") of the disease process, and established a repository of biological samples collected from the study participants.

In Benatar's new follow-up study, the original group of 30 participants will expand to include presymptomatic individuals with mutations in other ALS susceptibility genes such as TDP43 and FUS.

Study aims include better definition of the presymptomatic stage of familial ALS, identification of environmental factors that might modify the age of disease onset among genetically susceptible individuals, continued development of biomarkers of early disease, and expansion of the existing collection of biological specimens.

Benatar's exploration into the presymptomatic phase of ALS could facilitate earlier recognition and diagnosis of both the familial and sporadic forms of the disease, which in turn could point the way toward therapeutics aimed at prevention or delay of ALS onset, as well as attempts to stop or slow the disease before it causes irreversible damage.

"MDA has shown great foresight in recognizing the importance of this study and the kind of long-term contribution that it can make to our understanding of the disease," Benatar said.

Funding for this MDA grant began August 1, 2010.

MDA awarded $408,915 to Carmen Bertoni, assistant professor of neurology at the University of California, Los Angeles, to continue research into DNA repair strategies in Duchenne muscular dystrophy (DMD).

Bertoni's research group has pioneered a technology that uses small molecules called single-stranded oligonucleotides, or ssODNs, to act directly on the source of the problem in DMD: the "genetic blueprint," or DNA. In people with DMD, the DNA that makes up the dystrophin gene contains errors that lead to missing or nonfunctional dystrophin protein.

"We use ssODNs to let the muscle know of those errors and give the opportunity to each cell that composes muscles to correct the mistake," Bertoni said. The fix is permanent, negating the need for ongoing treatment.

The process involves injection of the oligonucleotide into muscle, where it seeks the flawed region of DNA and highlights the specific error. The cell is alerted to the presence of the genetic defect, and activates the repair process to correct the flaw.

To date, the main limitation of ssODN-mediated gene correction has been the low frequency of gene repair. Bertoni's group aims to increase the strategy's efficiency to generate levels of gene repair that are clinically meaningful as a viable treatment for DMD.

The group will test the oligonucleotides first in muscle cells and then, once investigators have optimized the structure, in dystrophin deficient mice.

"MDA has had and continues to have a pivotal role in the development of this technology for the treatment of DMD," Bertoni said. "Without any question, the advance of clinical applications to DMD using ssODNs heavily relies on the help that continues to be received by the MDA. It is a great honor to be part of the numerous and extremely talented scientists that are supported by the Association."

Funding for this MDA grant began August 1, 2010.

Michele Calos, professor in the department of genetics at Stanford University School of Medicine in Stanford, Calif., received a $200,000 MDA research grant to develop a new stem cell-based therapy for Duchenne muscular dystrophy (DMD).

"Funding from MDA is critical for us to develop these new ideas that could lead to novel strategies to combat DMD," Calos said. "A prior MDA grant has allowed us to make progress in the studies, and we are now poised to perform the critical experiments."

The new work involves extraction of stem cells from DMD mice. A good copy of the dystrophin gene, necessary but missing in DMD, will be added to the cells, which will then be returned to the mouse where they are expected to home in on damaged muscle and help create new muscle fibers to replace those damaged by the disease. The investigative team will also conduct experiments using a type of adult human stem cell called "adipose-derived mesenchymal stem cells," which are purified from fat tissue and have been shown to develop into muscle.

Successful results in these experiments eventually may lead to clinical trials and may indicate potential for use of a similar approach in other muscle diseases.

"Progress in the development of stem cell therapies offers major hope for regeneration of muscle mass and strength," Calos said, even for those in whom the disease has progressed.

Funding for this MDA grant began August 1, 2010.

MDA awarded $330,000 to professor of medicine Ju Chen at the University of California, San Diego, for research into the role of a protein called Cypher in skeletal muscle function and disease.

Mutations in Cypher result in myofibrillar myopathy (MFM) and late-onset distal myopathy, and results from completed studies in people with myotonic muscular dystrophy (MMD, or DM) indentified flawed Cypher isoforms (different forms of the same protein) in skeletal muscle tissues. Cypher activity also has been shown to be significantly decreased in mice exhibiting skeletal muscle atrophy.

"These observations suggest that Cypher plays essential roles in skeletal muscle function and disease," Chen said. "A better understanding of the function of Cypher in its various forms is key to developing therapies for Cypher-based MFM and potentially other myopathies."

In its work to understand the role of Cypher in skeletal muscle function and gain insight into the mechanisms by which mutations in Cypher cause skeletal muscle myopathy, the Chen research team will conduct experiments using three different genetically manipulated mouse models: an adult mouse lacking Cypher in skeletal muscle; mice lacking a short form of Cypher; and mice lacking a long form of the protein. Analysis will include comprehensive biochemical and functional evaluation, as well as evaluation of the microscopic structure of cells in skeletal mouse muscle tissue.

"Without funding from MDA, it would not be possible to perform these important studies," Chen said.

Funding for this MDA grant began August 1, 2010.

MDA awarded a grant totaling $160,000 to Dawn Cornelison, assistant professor in the division of biology at the University of Missouri-Columbia, for research into the role of a protein called syndecan-4 in muscle tissue repair after damage.

After muscle injury, Cornelison said, satellite cells (immature muscle cells) move in to make repairs. In order to do their job, the satellite cells need to perform a series of activities: They must first be 'activated' by signals resulting from injury; then divide several times to make more potential muscle cells; then stop dividing; and finally mature, or differentiate, into functional muscle.

In previous MDA-supported work, Cornelison and her research team analyzed a mouse engineered to lack the syndecan-4 protein and found that its satellite cells were unable to perform all of their normal muscle-repair functions. The group found that different parts of the syndecan-4 protein are responsible for regulating different satellite-cell functions in the muscle-repair process.

In its new work the group aims to identify proteins that interact with syndecan-4 and gain a better understanding of how the satellite cells carry out the four activities necessary for muscle repair.

"Funding from MDA has been critical to the progress and development of both this project and my scientific career," Cornelison said. "Particularly in the current environment, with the odds of obtaining funding for general research falling below 10 percent, knowing that MDA is there to sponsor research into muscle disease and regeneration is critical to retaining scientists in the field and encouraging others to undertake research into muscle diseases."

Funding for this MDA grant began August 1, 2010.

MDA awarded a grant totaling $339,561 to Richard Cripps, professor and chair of biology at the University of New Mexico in Albuquerque, for research into the role of the Trim32 gene in type 2H limb-girdle muscular dystrophy (LGMD).

The Trim32 gene carries instructions for the important Trim32 protein, required by muscle cells at the right times and in the necessary amounts for muscles to remain healthy and stable.

Cripps and colleagues currently are working to establish a model system in which to study the fruit fly version of Trim32.

"Our research will address fundamental questions about how this gene and its protein specifically behave in both the healthy and muscular dystrophy disease states of muscle," Cripps said. "Finding the answers to these questions will help researchers find critical points of intersection for the future treatment of this form of limb-girdle muscular dystrophy."

In experiments involving the fruit fly model, the team aims to identify the roles and behavior of Trim32 in muscles, including the structures it forms, the role these structures play in tissue stability, and interaction between the Trim32 protein and any other proteins.

"Having been funded by MDA off and on since I was a postdoctoral fellow in 1991-1992, I can deeply appreciate the support that MDA provides to scientists and fellows," Cripps said. "There is a greater willingness in MDA to take a chance on research that is of a higher risk, and the value of this approach has been borne out by the many achievements that MDA funding brings."

Funding for this MDA grant began August 1, 2010.

MDA awarded $179,327 to scientific researcher Sean Forbes at the University of Florida in Gainesville for research into impaired blood flow to muscles lacking the dystrophin protein in Duchenne muscular dystrophy (DMD).

Forbes plans to study an enzyme called neuronal nitric oxide synthase (nNOS) that contributes to blood flow and delivery of oxygen to muscles in the mdx research mouse model of DMD and another mouse model lacking nNOS. He’ll test the hypothesis that impaired blood flow during and following muscle use leads to inadequate delivery of oxygen to the muscles in DMD, contributing to muscle damage and accelerating the loss of healthy muscle tissue.

Forbes and his team will attempt to improve muscle blood flow via administration of the drug sildenafil citrate (brand name Viagra) and observe whether it lessens damage in DMD muscles. The group will use magnetic resonance imaging (MRI) and another type of imaging called spectroscopy (MRS) to measure changes in muscle blood flow, energy production and damage in mice. These noninvasive techniques may prove to be sensitive tools to monitor and visualize disease progression and effectiveness of treatment in DMD.

"This MDA Development Grant provides a valuable opportunity to pursue this exciting and important area of research. The funding will enable us to perform studies to examine muscle metabolism and blood flow in dystrophy, and will examine the effects of a prospective treatment for DMD," Forbes said. "These studies have the potential to set the foundation for future studies in children with DMD."

Funding for this MDA grant began August 1, 2010.

Michael Francis, associate professor in the department of neurobiology at the University of Massachusetts Medical School in Worcester, received an MDA grant totaling $330,000 for research into effects on the connection between nerve and muscle known as the neuromuscular junction, or NMJ, in congenital myasthenic syndrome (CMS).

Francis and colleagues have engineered a research model using C. elegans, a type of worm also known as a nematode, that mimics many of the key features of a form of CMS called slow-channel congenital myasthenic syndrome (SCCMS). The team will use sophisticated genetic tools paired with measurements of the electrical properties such as voltage changes and electric currents in cells and tissues, and a sophisticated imaging technique called confocal microscopy in its analysis of NMJ function in both normal and diseased tissues.

The investigators plan also to pinpoint the molecular mechanisms underlying degeneration and alterations in NMJ function caused by neuromuscular disorders.

"We expect our efforts will provide key mechanistic insights into conserved molecular pathways important for NMJ development and function in the normal organism, as well as during diseases that impact NMJ function, such as SCCMS," Francis said. "Identifying the components of these pathways will be important for the development of new therapeutic strategies to combat neuromuscular disorders."

Funding for this MDA grant began August 1, 2010.

MDA awarded $180,000 to research scientist Brian Freibaum at St. Jude Children's Research Hospital in Memphis, Tenn., for research into the mechanism by which toxic TDP43 protein leads to the development and progression of some forms of ALS (amyotrophic lateral sclerosis, or Lou Gehrig's disease).

Prior research has suggested that mutant (flawed) TDP43 protein is a critical component in the development of some forms of ALS, although the mechanism by which it works remains unknown. Freibaum has planned a three-tiered approach to uncover the protein's role in the disease.

First, Freibaum's team will test for toxicity in various tissues of fruit flies engineered to carry the human TDP43 gene. Using different mutant (flawed) and shortened forms of the protein is expected to help pinpoint what parts of the TDP43 protein are required for disease progression.

Next, the team will use human cell lines and mouse motor neurons (nerve cells that control muscle) to explore how TDP43 interacts with other proteins, where it normally localizes in the cell, and whether these locations are altered with mutant forms of the protein. Further experiments will be used to determine what other proteins interact with normal and flawed TDP43 protein.

Finally, the group will use the fruit fly model to perform genetic screens designed to explore how other proteins cooperate with mutant TDP43 to induce toxicity in motor neurons.

Greater understanding of the role TDP43 plays in ALS will help provide a more targeted approach to the development of new therapies.

"I am honored to be funded by such a prestigious organization that has a strong history in the advancement of neuromuscular disease research," Freibaum said. "This funding will be of great significance to our lab’s ALS research."

Funding for this MDA grant began August 1, 2010.

Renzhi Han, assistant professor of physiology at Loyola University Medical Center in Maywood, Ill., received an MDA grant totaling $405,000 to study mutations in the dysferlin gene that lead to development of several types of muscle diseases known as "dysferlinopathies," including type 2B limb-girdle muscular dystrophy (LGMD).

Previous studies have shown that inflammation, a component of the "innate immune system," is prominent in muscle samples deficient in dysferlin protein. (The innate immune system specializes in providing immediate defense against infection in the body. By contrast, the adaptive immune system specializes in building up protective immunity.)

Han speculates that an attack carried out by the innate immune system might play a role in the onset and progression of muscle diseases that result from dysferlin mutations, and that disabling this immunological response may ameliorate the symptoms of muscular dystrophies caused by dysferlin deficiency.

The investigators will use research mouse models deficient in both dysferlin and the innate immune system to test their hypothesis. The team also will study whether increasing the activity of a biochemical inhibitor of the immune system has any effect on disease progression in dysferlin-deficient mice.

"This research will enable us to accomplish the critical step in our goal to design a therapy for dysferlin-deficient muscular dystrophy," Han said. "This work could not go further without the support of MDA."

Funding for this MDA grant began August 1, 2010.

MDA awarded $374,511 to Oliver Hobert, professor of biochemistry and molecular biophysics at Columbia University, New York, to study the function of the TDP43 gene, mutations in which can cause ALS (amyotrophic lateral sclerosis, or Lou Gehrig's disease).

"In order to diagnose and treat ALS, it is important to understand the molecular events that underlie this disease," Hobert said. The TDP43 gene, known to cause ALS in humans, "works in a manner that is not understood. Our goal is to better understand the function of this gene."

Using the invertebrate C. elegans (nematode, or roundworm) model, Hobert's team will study the TDP43 gene in order to determine its function and possible interaction with other genes.

"Funding by the MDA means a great deal to us — not just because of the financial support but also because it motivates us to direct our basic research toward questions that are medically relevant," Hobert said.

Funding for this MDA grant began August 1, 2010.

Research scientist Dena Jacob at Thomas Jefferson University in Philadelphia, received an MDA grant totaling $180,000 for research into decreasing cells' resistance to therapeutic medications in ALS (amyotrophic lateral sclerosis, or Lou Gehrig’s disease).

Jacob plans to study a "transporter" protein called P-glycoprotein, or P-gp, which belongs to a class of "transporter" proteins that pump drugs out of cells. P-gp recognizes a broad range of drugs and normally is found in cells of the blood brain and blood spinal cord barriers and also in some affected brain tissue, limiting drug penetration.

"P-gp expression in the spinal cord itself may impart overt pharmacoresistance (drug resistance) to ALS-affected cells and account for the poor therapeutic effect observed in [a number of previous] ALS clinical trials," Jacob said.

Jacob's findings could lead to re-examination of many previously unsuccessful clinical drug trials and encourage the development of new, combined therapeutic strategies for ALS.

"I am extremely honored and excited to be a Development Grant recipient of the MDA, an organization whose efforts and support are essential for advancing our understanding of neuromuscular diseases," Jacob said. "This funding will enable me to complete important experiments that will help us identify improved therapeutic strategies to target the mouse model of ALS and, ultimately, humans with this disease."

Funding for this MDA grant began August 1, 2010.

MDA awarded Professor Bernard Jasmin, vice-dean of research at the University of Ottawa in Canada, a grant totaling $360,000 to study the impact of small molecules called "exercise mimetics" in Duchenne muscular dystrophy (DMD).

One approach to counteract the effects of the loss of the muscle protein dystrophin in DMD involves increasing levels of a similar structural protein called utrophin. Studies performed in a widely used mouse model of DMD, the mdx mouse, have shown that increased utrophin expression can compensate for the lack of dystrophin and ameliorate symptoms of the disease.

Because utrophin activity normally is low and focused only in certain regions of the muscle, Jasmin's laboratory looks for ways to increase levels of the protein in DMD muscles.

"Our first preclinical study showed that treatment of mdx mice with an exercise mimetic known as GW501516 resulted in an increase in utrophin expression, improved muscle function and improved pathological symptoms of the disease," Jasmin said.

His current work builds on the previous study results, with plans for continued testing of GW501516 in the mdx mouse model of DMD, and expansion of the research to include testing of other molecules including AICAR, beta-GPA and resveratrol.

Because of the possible effects of exercise mimetics on utrophin in DMD-affected muscles, the small molecules hold potential for dramatic improvement in quality of life for individuals with the disease. GW501516 currently is under development to treat obesity by the pharmaceutical company GlaxoSmithKline, which could help speed its development for therapeutic use in people with DMD.

"MDA funding has allowed us to take a relatively basic molecular observation and perform research that eventually could result in development of novel therapeutic approaches for treatment of people with DMD," Jasmin said.

Funding for this MDA grant began August 1, 2010.

MDA awarded a grant totaling $282,630 to Albena Jordanova, professor in the department of genetics at the University of Antwerp, Belgium, for research into the molecular causes of, and potential drug targets for, a recently discovered form of Charcot-Marie-Tooth (CMT) disease known as dominant intermediate CMT type C (DI-CMTC).

Jordanova's research team was the first to describe the new CMT type and showed that the disease is caused by different mutations (genetic flaws) in the gene that carries instructions for an enzyme called tyrosyl-tRNA synthetase (YARS).

In their new work, Jordanova and colleagues aim to identify genes that modify the neurodegenerative symptoms of DI-CMTC present in a fruit fly research model of the disease. The team will then test the modifying genes to determine how well each is able to interact with drug-like chemical compounds, in an effort to pinpoint the best drug targets for DI-CMT and other types of the disease.

The "best gene hits" will be those that carry instructions for proteins that are both structurally favorable — that is, their protein folds are amenable to interactions with drug-like chemical components — and functionally favorable with a proven ability to affect the DI-CMTC disease process.

"MDA funding is absolutely essential to start this work," Jordanova said. "This is important seeding money for us to initiate a midterm research program and to perform the research we dream of."

Funding for this MDA grant began August 1, 2010.

MDA awarded a grant totaling $345,000 to Jean-Pierre Julien, professor at Laval University, Canada, for research into genetic variations in a protein called chromogranin B (CHGB) that has been shown to modify disease risk and hasten onset in a type of familial ALS (amyotrophic lateral sclerosis, or Lou Gehrig's disease).

In previous studies, Julien's group discovered that CHGB interacts with mutant, or flawed, forms of the superoxide dismutase 1 (SOD1) protein associated with SOD1-mediated familial, or inherited, ALS.

The team studied variations in the chromogranin B gene, which carries the instructions for the CHGB protein, in a large group of people with and without ALS. Results showed that people with ALS were twice as likely to have a common CHGB variant called P413L than those without the disease. The P413L CHGB variant also correlated with a 2.2-fold greater relative risk for development of ALS, and among those with the disease, onset of symptoms occurred an average 10 years earlier for those with the variant than for those without it.

In its new work, Julien's group will conduct experiments in cultured nerve cells and in a strain of mice engineered to carry the human CHGB variant to test the hypothesis that P413L increases vulnerability of motor neurons. The investigators also aim to determine whether the variant affects CHGB's interaction with mutant SOD1 and uncover the exact mechanism by which it increases risk of ALS and modifies disease onset.

"MDA funding is timely and needed," Julien said, "to determine how this particular genetic variant increases risk of ALS and hastens disease onset."

Funding for this MDA grant began August 1, 2010.

Virginia Kimonis, chief of the division of genetics and metabolism at the University of California, Irvine, received an MDA grant totaling $372,000 to conduct experiments designed to uncover the underlying molecular cause of inclusion-body myopathy associated with Paget's disease of Bone and/or frontotemporal dementia (IBMPFD).

In 2004, Kimonis and colleagues published study results describing the rare disease IBMPFD and identifying the gene responsible for it — valosin-containing protein, or VCP. The group also created a mouse model that carries a flawed human VCP gene and exhibits symptoms similar to those in humans with IBMPFD, including decreased muscle strength and coordination, fatigue, enlarged vacuoles (bubblelike spaces) in the quadriceps muscles, swelling of cellular "energy factories" called mitochondria and abnormal protein clumps.

The Kimonis lab's new project includes the creation of a new mouse model aimed at providing proof of principle that amelioration of IBMPFD symptoms is possible. The team will evaluate functional and biochemical indicators to assess disease progression and effects in these mice. The group also will study whether physical exercise accelerates or decelerates the progression of muscle weakness in the mice, as a means of assessing what effects physical exercise may have on people with IBMPFD.

"Funding from MDA is critical for our laboratory to be able to continue this important work in understanding the key mechanisms related to this progressive and fatal muscle disease," Kimonis said. "Because of its similarities to other muscle disorders such as limb-girdle muscular dystrophy (LGMD), facioscapulohumeral muscular dystrophy (FSHD) and ALS (amyotrophic lateral sclerosis, or Lou Gehrig's disease), it's hoped that this research will advance a deeper understanding of mechanisms and potential treatments across a broad spectrum of neuromuscular diseases."

Funding for this MDA grant began August 1, 2010.

MDA awarded associate professor Ashok Kumar at the University of Louisville School of Medicine in Louisville, Ky., a grant totaling $349,206 for continued study of the molecular mechanisms that underlie disease onset and progression in Duchenne muscular dystrophy (DMD).

Kumar's team will work to identify biochemical changes in skeletal muscle caused by the loss of functional dystrophin protein in the disease.

"Recent studies in my laboratory have identified a set of proteins known as 'matrix metalloproteinase' (MMPs) that are highly deregulated in dystrophic muscles," Kumar said. "Interestingly, MMPs are the major enzymes responsible for tissue degradation in a number of disease states and conditions."

Kumar and colleagues have found that treatment with a molecule called batimastat is highly effective at ameliorating disease symptoms such as fiber necrosis (death) and inflammation, and improving skeletal muscle structure in dystrophin-deficient mice.

Now, the team will investigate the therapeutic potential of MMP-inhibiting drugs, some of which are already being used for other diseases in humans (including chemical relatives of the antibiotic tetracycline).

"I strongly believe that the current funding provided by MDA will open new avenues for treatment of DMD in the near future," Kumar said. "I sincerely thank MDA for its generous funding to pursue my research work in DMD."

Funding for this MDA grant began August 1, 2010.

MDA awarded a grant totaling $375,000 to Louis Kunkel, director of the program in genomics at Children's Hospital in Boston, for research into compounds already approved for human use that may alter disease progression in Duchenne muscular dystrophy (DMD).

"There are thousands of compounds developed for use in humans for a variety purposes such as pain, diabetes and heart disease," Kunkel said. "The vast majority of these compounds have never been tested for their possible use in other diseases such as muscular dystrophy, something that can be easily accomplished in zebra fish."

In previous studies, Kunkel's lab has identified several compounds that increase the survival rate in zebra fish with a DMD-like disease and that are approved by the U.S. Food and Drug Administration for use in humans.

The team now plans to search for additional compounds, which it will test in the zebra fish model. The investigators will analyze indicators including survival rates, muscle function and toxicity, to determine the best candidate to go forward into mouse models.

"The beauty of the zebra fish is that they are permeable," Kunkel said. "They present an opportunity to directly test compounds in a live organism with the disease due to their large numbers, small size, transparency and their ability to absorb chemicals from their water environment."

Funding for this MDA grant began August 1, 2010.

MDA awarded Albert La Spada, chief of the division of genetics in the department of pediatrics at the University of California, San Diego, $330,000 to study what causes nerve cells called motor neurons to die in spinal-bulbar muscular atrophy (SBMA) and other neurodegenerative diseases such as ALS (amyotrophic lateral sclerosis, or Lou Gehrig's disease) and spinal muscular atrophy (SMA).

La Spada and colleagues have created mouse and motor neuron cell-culture models of SBMA, which results from a genetic flaw in the androgen receptor (AR) gene. Completed studies have led to the identification of candidate pathways that may spur motor neuron degeneration.

Now, La Spada's team will focus on the androgen receptor (AR) protein, which is biochemically modified and made toxic in SBMA. An understanding of the particular proteins, or "enzymes," that catalyze such chemical modifications will set the stage for identification of drug compounds designed to inhibit them.

The team also will work to identify other cell types involved in driving the SBMA disease process. Using a specialized mouse model, the investigators will "turn off" activity of the mutant, or flawed, AR protein in different cell types, starting with motor neurons and muscle cells, in order to determine whether those cells contribute to making motor neurons sick.

"MDA funding has been critical in allowing us to aggressively pursue cutting-edge research approaches in my lab," La Spada said. "Without MDA, there is no way that we could have made all the progress that we did over the last decade."

Findings from the new work may identify targets for the development of therapies to treat SBMA, ALS and SMA.

Funding for this MDA grant began August 1, 2010.

MDA awarded $375,000 to Gordon Lynch, professor of physiology at the University of Melbourne, Australia, for research into strategies aimed at improving muscle function in Duchenne muscular dystrophy (DMD).

Lynch's research builds on previous studies conducted by his group and others that have demonstrated the potential of proteins called "growth factors" to preserve muscle fibers. These growth factors are regulated by a family of proteins called insulin-like growth factor binding proteins (IGFBPs).

Lynch's team has uncovered evidence that a particular IGF binding protein called IGFBP-2 may play a key role in the cycles of muscle fiber damage and repair that occur in DMD.

Using a mouse model genetically altered to exhibit either increased or decreased IGFBP-2 activity, the team will seek to determine whether the protein aggravates or improves the DMD disease process. The researchers also will study the effects of IGFBP-2 and other IGF binding proteins on muscle development and muscle repair.

"The MDA has generously supported my laboratory’s research for over a decade, allowing my team to make consistent and significant contributions to the field, including demonstrating the exciting potential of growth factors for improving muscle function in muscular dystrophy," Lynch said. "Support from the MDA has been critical in facilitating my mentorship of some very talented young doctoral students and postdoctoral scientists who have each made important contributions to muscular dystrophy research and become successful independent investigators in their own right."

Lynch's new research will generate important new information about the functional roles of IGF binding proteins in muscular dystrophy, and contribute to the development of therapies designed to improve the quality of life for those with muscle disease.

Funding for this MDA grant began August 1, 2010.

MDA awarded a grant totaling $79,277 to John Manfredi, chief scientific officer at Sfida BioLogic Inc., in Salt Lake City, Utah, for continued research into new drug compounds that promote the growth and function of motor neurons (nerve cells), and that may have potential as therapeutics for treatment of spinal muscular atrophy (SMA).

Manfredi's research team aims to determine the therapeutic potential of their compounds using a zebra fish research model that has been genetically engineered to simulate aspects of SMA. Results will determine whether the drugs merit future testing in more sophisticated and expensive models of the disease.

The compounds identified by Manfredi and colleagues "exhibit attractive pharmaceutical properties," Manfredi said. "They are not toxic; they penetrate the central nervous system; they exhibit appropriate half-lives in tissues; they can be administered orally; their permeability and solubility characteristics are drug-like; and they can be economically synthesized."

The group also will test a sample of their compounds alongside a collection of biochemically similar, commercially available drugs. If the commercial drugs show effects in the zebra fish model of SMA, the results will support the hypothesis that their SMA-relevant effects are due to shared biochemical activity with the investigators' novel drugs. Positive results also could lead to development of the commercial compounds as SMA therapeutics.

"The funding provided by MDA is absolutely critical for evaluating the potential of our company’s proprietary compounds to treat spinal muscular atrophy," Manfredi said. "Indeed, if these or derivative compounds prove to be efficacious for the treatment of SMA, MDA can legitimately claim responsibility for their success."

Funding for this MDA grant began August 1, 2010.

Fernando Morales, head of the genetic section at the Health Research Institute of the University of Costa Rica, has received an MDA grant totaling $366,210 over three years. The funding will help support Morales’ research to define the molecular underpinnings of type 1 myotonic dystrophy (MMD1, or DM1) and factors that modify the course of this highly variable, multisystem disease.

The MMD1-causing mutation, an expansion in the DNA of a gene on chromosome 19, changes over time, usually becoming larger during a person's lifetime and when being passed between generations. In general, the larger the DNA expansion, the more profound the effects of it are likely to be. However, the correlation between disease severity and age of onset and size of the mutation isn't perfect.

In this project, Morales and colleagues aim to analyze how the MMD1 mutation changes over time and how the changes relate to the clinical course of the disease. They also plan to identify genetic and other modifiers of the MMD1 mutation.

"Importantly," said Morales, "these modifiers of the MMD1 mutation could also be modifiers of the disease, and if so, they could be used as therapeutic targets to delay disease onset and progression."

Morales said he hopes the project will generate more reliable genetic data that can provide more accurate information about prognosis to families and improve disease management and quality of life.

"Funding by MDA is of great importance to our project," Morales said. "Thanks to it, we can investigate in more detail several biological and clinical aspects related to MMD1 in which there is still controversy."

Funding for this MDA grant began August 1, 2010.

MDA awarded a grant totaling $375,000 to Francesco Muntoni, professor of pediatric neurology at University College London (UCL), United Kingdom, for research into the molecular mechanisms underlying central core disease (CCD) and multiminicore myopathies. Muntoni and Michael Duchen, professor of physiology, and cell and developmental biology, also at UCL, will work together, focusing on the mechanisms that lead to muscle weakness in the two diseases.

CCD and multiminicore myopathies both are caused by mutations in the ryanodine receptor gene (RYR1), which carries instructions for the RYR1 protein.

"While we have known for a number of years that RYR1 gene mutations were associated with these core myopathies, the precise mechanism leading to muscle weakness in affected individuals is less clear," Muntoni said.

It's known that RYR1 plays a key role in the release of calcium in skeletal muscle; the calcium release is followed by muscle contraction.

Muntoni and Duchen will investigate the hypothesis that chronic calcium dysregulation due to dysfunctional RYR1 protein leads to malfunction, in each muscle fiber, of the cell's energy factories, called mitochondria. With muscle cells from individuals carrying RYR1 mutations, the two will study various aspects of mitochondria health and function following exposure to abnormal calcium release.

"This study is a follow-on of a currently funded MDA grant that is coming to an end," Muntoni said. "The current MDA grant allowed us to improve our insight on the correlation between different mutations in RYR1 and the core myopathies. Without the original MDA grant, we would not have been in a position to establish these novel correlations between RYR1 and congenital diseases."

An improved understanding of the link between dysregulated calcium handling and mitochondria dysfunction could lead to new therapeutic interventions for individuals with core myopathies.

Funding for this MDA grant began August 1, 2010.

Josephine Nalbantoglu, associate professor in the department of neurology and neurosurgery at McGill University in Montreal, Canada, received an MDA grant totaling $313,170 for research into increasing levels of the muscle protein utrophin as a therapeutic strategy in Duchenne muscular dystrophy (DMD).

Utrophin is structurally similar to the crucial muscle protein dystrophin, missing in DMD. Nalbantoglu and colleagues aim to validate, in muscle cells and in an animal model of DMD, the potential for utrophin to fulfill some of the functions normally carried out by dystrophin and compensate for the missing protein.

In completed studies, Nalbantoglu's group has shown in an animal model of DMD that utrophin levels can be increased by targeting specific sites in the gene that carries the instructions for utrophin with proteins called transcription factors. Muscle injected with these proteins exhibited fewer hallmarks of disease and increased function despite the absence of dystrophin.

Using the same approach, the team will now refine transcription factors designed to result in increased levels of the human utrophin protein. In parallel, the investigators will explore various methods for delivering the factors throughout the body.

"Funding from MDA is crucial for this work," Nalbantoglu said. The approach of using artificial transcription factors to increase levels of specific genes would have wide application to other muscle diseases. Artificial transcription factors are already in clinical trials for some non-muscle diseases."

Funding for this MDA grant began August 1, 2010.

Vihang Narkar, assistant professor at the University of Texas Health Science Center at Houston, was awarded $302,326 to study the potential therapeutic value of increasing the overall amount of a specific type of muscle called "aerobic muscle" in Duchenne muscular dystrophy (DMD).

Loss of the structural protein dystrophin, the underlying cause of DMD, causes muscle instability, damage and progressive weakness, accompanied by a decrease in the ability to produce energy, along with severe fatigue.

Because aerobic muscles have enhanced capacity for energy production (which takes place inside cell compartments called mitochondria) and are resistant to fatigue, it's thought that biological regulators that could stimulate production of this type of muscle in individuals with DMD might ameliorate some of the symptoms in the disease.

Narkar's group has discovered one such "bio-regulator," a protein called ERRgamma. When activated in the skeletal muscles of mice with a DMD-like disease, ERRgamma prompts production of a highly aerobic and fatigue-resistant muscle.

In their new work, Narkar and colleagues plan to genetically activate ERRgamma in the skeletal muscles of mdx mice, a rodent model of DMD, as a means of increasing the overall amount of aerobic, fatigue-resistant muscles. A sophisticated battery of tests will help the investigators evaluate the effects of ERRgamma on muscle health and determine whether targeting the protein to dystrophic muscles may have therapeutic benefit in DMD.

"This is our first research grant from the Muscular Dystrophy Association," Narkar said. "It will play a very critical role in initiating and advancing this new and therapeutically promising area of research."

Funding for this MDA grant began August 1, 2010.

Daniel Offen, head of the neurology laboratory at Tel-Aviv University, Israel, received an MDA grant totaling $359,700 for research into a combined cell and gene therapy approach for ALS (amyotrophic lateral sclerosis, or Lou Gehrig's disease).

Offen's research team will engineer progenitor cells, a type of immature cell that forms muscle, to express various combinations of neurotrophic factors (proteins that support motor neuron health and which previously have been reported to be beneficial in rodent models of ALS). The investigators will inject mixtures of progenitor cells expressing different combinations of the neurotrophic factors into muscles in the SOD1 research mouse model of ALS and then monitor the course of the disease.

Behavioral, physical and biochemical survival indicators will be observed in order to analyze the effects of the stable activity of different growth factors on disease progression, extension of life span and quality of life in ALS. The experiments with the most promising results will then be duplicated using immature human muscle cells.

It's hoped the work will lead to a new cell and gene therapy strategy for treatment of human ALS.

"Past MDA support has enabled us to develop several approaches to isolate and propagate myogenic cells in cultures, some of which will be used in the present investigation," Offen said. "Our current MDA award will enable us to conduct the proposed project and hopefully to contribute to the treatment of ALS patients."

Funding for this MDA grant began August 1, 2010.

MDA awarded $312,699 to Leo Pallanck, associate professor of genome sciences at the University of Washington, Seattle, for research into elimination of flawed cell machinery that is the underlying cause of mitochondrial myopathy.

Mitochondria, the tiny "power plants" responsible for producing nearly all of the energy needed for a human cell to function properly, have their own genes — stretches of DNA that carry the genetic instructions used in protein production. There can be hundreds of mitochondria in a single cell, and large cells with particularly high energy demands, such as skeletal and cardiac (heart) muscle cells, may harbor more than 10,000.

Mutated, or flawed, mitochondrial genes lead to flawed mitochondrial proteins that impair the ability of the mitochondria to produce energy and, by extension, the ability of the cell to function.

Recent studies have shown that have a pair of proteins known as PINK1 and Parkin can recognize damaged mitochondria and promote their destruction.

Pallanck and his research team aim to test the hypothesis that PINK1 and Parkin, in conjunction with other cellular factors, can detect and selectively destroy flawed mitochondria, thus alleviating the symptoms associated with mitochondrial mutations by keeping numbers of the defective mitochondria low.

In experiments on a fruit fly model, the investigators will vary the amounts of PINK1 and Parkin, and study the effects on the frequency of mitochondrial mutations and their associated symptoms, including degeneration of muscles and nerves. The work could lead to the development of strategies that can be used to reduce the overall amount of mutation-bearing mitochondria in humans.

"This is my first grant from the Muscular Dystrophy Association, and I cannot overstate the importance of this funding to our work," Pallanck said. "We couldn't perform these studies without support from the MDA."

Funding for this MDA grant began August 1, 2010.

MDA awarded $358,653 to Wilfried Rossoll, assistant professor at Emory University in Atlanta, for research into the effects on nerve cells, or "motor neurons," of toxic TDP43 protein, implicated in ALS (amyotrophic lateral sclerosis, or Lou Gehrig's disease).

"Recently, we have developed novel tools to study protein localization, trafficking and function in cultured primary motor neurons," Rossoll said. "Funding from MDA will now allow us to apply these methods to address the biological role of the ALS disease protein TDP43 in motor neurons."

TDP43 protein is present in all body tissues and serves multiple roles in the processing of messenger RNAs (mRNAs), the molecules that carry genetic information from DNA to the sites of protein synthesis. However, the function of TDP43 in motor neurons and vulnerability of those cells to the protein still is not understood.

"Most studies on the role of TDP43 protein have used non-neuronal cell lines, so its function in the highly specialized motor neuron cell type remains unclear," Rossoll said. "We will use cell cultures of primary motor neurons and motor neurons generated from stem cells to study the function of TDP43 in this cell type, and to gain information about its involvement in ALS."

Preliminary data generated by Rossoll's team suggests that TDP43 plays a role in mRNA processing in the long axon fibers that conduct electrical impulses to muscle cells. The team will test the hypothesis that decreased levels of TDP43 in mRNA-protein complexes in these axons may contribute to the degeneration of motor neurons in ALS.

Increased understanding of the molecular and cellular functions of TDP43 could help lead to the development of therapies that delay or even prevent motor neuron degeneration in people with ALS or other neurodegenerative diseases.

Funding for this MDA grant began August 1, 2010.

MDA awarded $180,000 to Ji-Yeon Shin, a research scientist at Columbia University Medical Center, New York, for continued study of the molecular mechanisms that underlie Emery-Dreifuss muscular dystrophy (EDMD).

"EDMD is an inherited disease of skeletal muscle and heart. Although diagnosis has been improved by the discovery of the most common genetic mutations that cause EDMD, we still have a poor understanding of how these mutations cause muscular dystrophy," Shin said.

Shin's lab previously has discovered that proteins produced from instructions carried by the mutated genes in most cases of EDMD interact with another protein, called LAP1 (lamina-associated polypeptide 1), whose function as yet remains unknown. Shin's team has engineered a new "muscle specific LAP1 knockout" mouse that exhibits symptoms that mimic those in EDMD and has potential therefore to be used as an EDMD research mouse model.

In addition to further characterizing the mouse, Shin's new work will include investigation into how the LAP1 protein affects muscle cell function in mouse and human cultured cells, and in living mice.

The team hopes to generate a better understanding of how specific genetic mutations cause EDMD, which should enable it to identify new processes, such as cell signaling pathways, that could be targets for the development of novel drugs to treat the disease.

"I am very happy to be selected as a Development Grant awardee, and I have great hopes for what I can achieve from the current project funded by MDA," Shin said. "The previous results that have been generated in the last two years built up exciting groundwork. Now the support of MDA will bring this project to the next level and allow us to study cellular mechanisms that underlie the development of Emery-Dreifuss muscular dystrophy."

Funding for this MDA grant began August 1, 2010.

MDA awarded a grant totaling $347,832 to Shanthini Sockanathan, associate professor of neuroscience at the Johns Hopkins University School of Medicine in Baltimore, for research into the molecular causes of nerve cell, or motor neuron, degeneration in diseases including ALS (amyotrophic lateral sclerosis, or Lou Gehrig's disease).

Sockanathan and colleagues will study the Gde2 mouse, a new model of motor neuron degeneration that they developed based on studies of the maturation, or "differentiation," of motor neurons in embryonic mouse spinal cords.

The Gde2 mouse lacks a protein called GDE2. Mouse embryos lacking the protein possess significantly decreased numbers of motor neurons, but at birth they contain numbers of these cells equivalent to those of control mice. As adults, the Gde2 mice experience a significant loss of motor neurons, with the remaining nerve cells indicating progressive degeneration.

In their new work, the investigators will study the physical and biological characteristics of Gde2 mice, and monitor the progression of motor neuron degeneration in older animals. Because the GDE2 protein normally is active in both differentiating and fully matured motor neurons, the team plans to remove it in mice at different times during development to determine at which stage or stages GDE2 loss causes degeneration.

"MDA's clear commitment to funding basic research projects has been instrumental in my ability to develop and maintain an active research program dedicated to understanding the mechanisms of neuronal differentiation," Sockanathan said. "Indeed, my first grant as an independent investigator was from MDA, and this funding helped me establish my research program and, importantly, provided me with the flexibility to chart a new direction in my work. Over the years, I have been fortunate to receive more funding from MDA, and in each case this funding has helped me broaden my research program in novel areas."

Funding for this MDA grant began August 1, 2010.

MDA awarded a research grant totaling $368,100 to professor Steve Wilton at the University of Western Australia in Perth, for continued research into a strategy called "exon skipping," which bypasses mutations in the dystrophin gene responsible for Duchenne muscular dystrophy (DMD).

Wilton and his team were one of the first groups to use exon skipping to bypass DMD-causing mutations and partially restore production of the dystrophin protein missing in the disease. Since then, the exon-skipping field has rapidly progressed from an idea first demonstrated in studies of isolated cells to application in animal models of the disease and, more recently, to human clinical trials.

Two types of exon-skipping drugs that have reached the clinical trial stage are called 2Omethyl and morpholino. A third drug, 2 methoxyethyl, or "MOE," appears to be well-suited for exon skipping but was not made available for study until recently.

In their new work, Wilton and his team will conduct detailed preclinical studies in the mdx research mouse model of DMD to assess MOE's suitability for exon skipping.

In addition, the group has developed a number of other drugs designed to bypass a variety of different dystrophin mutations that should be responsive to exon skipping. The investigators will study the effectiveness and side-effect profiles of the new drugs.

"Without MDA support, the exon-skipping field would not be anywhere near as advanced as it is," Wilton said. "MDA funded this research when many other research funding bodies were very skeptical."

Funding for this MDA grant began August 1, 2010.

MDA awarded $411,000 to Lee-Jun Wong, professor in the department of molecular and human genetics at Baylor College of Medicine, Houston, for research designed to produce a one-step diagnosis procedure for complex mitochondrial myopathies.

Diagnosis of mitochondrial disorders often is difficult because the disease can present with any of a number of different symptoms including muscle weakness, exercise intolerance, paralysis or weakness of the eye muscles, deafness, seizures, lack of coordination, movement disorder, and strokelike episodes.

Current diagnostic methods are based on the genetic testing of individual genes, one by one. Such a process can be tedious, expensive and time-consuming, and isn't able to uncover all the different types of mutations that may be responsible for mitochondrial disorders.

"The availability of a one-step diagnostic approach is particularly important since mitochondrial myopathy accounts for a large proportion of patients, from children to adults, with muscular dystrophy," Wong said. "Prompt, definitive diagnosis is essential for proper patient management, treatment and genetic counseling."

Wong's group plans to establish a one-step technology that will allow for the detection of all of the various types of mutations that can result in mitochondrial myopathy. The team will validate its technique using archived DNA samples from people with mitochondrial neuromuscular disorders.

"My first research grant was from MDA, and it helped me develop a comprehensive method to screen mutations in mitochondrial DNA," Wong said. "Now, as an established and experienced principal investigator focusing on the molecular diagnosis of mitochondrial disorders, new MDA funding allows me to bring in an innovative one-step approach to comprehensive diagnosis of the complex dual genome mitochondrial disorders."

Funding for this MDA grant began August 1, 2010.

MDA awarded a grant totaling $310,893 to Howard Worman, professor of medicine and pathology, and cell biology at Columbia University Medical Center in New York, for research into treatments that target the underlying cellular disease process responsible for heart damage, or "cardiomyopathy," in people with Emery-Dreifuss muscular dystrophy (EDMD).

In previous research supported by MDA, Worman's group showed that inhibiting proteins in the MAP kinase signaling pathway improved heart function in a mouse model of EDMD.

"The 'first-generation' inhibitors that we used are not suitable for humans," Worman said. "In the current research, we will study drugs of the same classes that have already been given to humans to determine if they similarly improve heart function in these mice."

The group plans to administer the new inhibitors and then examine the mice using some of the same methods doctors use to examine the heart in humans, including echocardiography (a type of imaging study used to assess the heart's pumping ability). The investigators will extend their findings to other EDMD mouse models known to develop cardiomyopathy, and then ascertain whether administration of MAP kinase signaling inhibitors causes toxicity in these mice.

Positive results generated by Worman's research could pave the way for clinical trials of MAP kinase signaling inhibitors to treat humans with EDMD.

"MDA has supported our research on Emery-Dreifuss muscular dystrophy for more than 10 years," Worman said. "This most recent grant will allow us to translate these past 10 years of basic laboratory research into a treatment for human patients with muscular dystrophy."

Funding for this MDA grant began August 1, 2010.

MDA awarded $375,000 to Daniela Zarnescu, assistant professor in neuroscience at the University of Arizona in Tucson, Ariz., to conduct gene and drug discovery research in a drosophila fruit fly model that carries a mutation in the TDP43 gene associated with a genetic form of human ALS (amyotrophic lateral sclerosis, or Lou Gehrig's disease).

Alterations in TDP43 gene activity in the fruit fly model lead to symptoms that mimic those found in humans with the disease, including the death of nerve cells called motor neurons and the formation of abnormal clumps of cellular material, called inclusions, that contain TDP43 proteins.

Zarnescu's group will conduct genetic screening tests on the flies in order to identify other genes that interact with TDP43, some of which may be involved in disease causation or progression. The group also will screen drugs in search of one or more that can rescue the defects in the fruit fly model.

The work could lead to novel therapeutic targets and approaches for ALS.

"Recognizing the need to fund such projects is not only an important step to ensure important discoveries in the field, but it also shows that MDA is at the forefront of the effort towards identifying new therapies and a potential cure for ALS," Zarnescu said.

Funding for this MDA grant began August 1, 2010.