The ALS Association’s TREAT ALS (Translational Research Advancing Therapies for ALS) Portfolio is a research endeavor enabling important global research to progress from the laboratory to the bedside. The focus of the program is to support novel ideas, build tools, partner with academia and industry to identify new potential therapies and support the infrastructure for clinical trials with the goal to find meaningful treatments for ALS and a cure.
The ALS Association is pleased to announce twenty new grants in laboratories throughout the United States, United Kingdom, Belgium, Italy and Spain. The new awards include four innovative discovery awards generously supported by The Alan Phillips Discovery Grant Award Fund and five Milton Safenowitz Postdoctoral Fellowship Awards. These new grants focus on understanding how the new genes TDP43 and FUS contribute to disease, development of a novel model system to understand neuronal activity, immune mechanisms in ALS and development of novel Riluzole (Rilutek) derivatives with improved activity.
In addition, investigators at a new biotech company, Ossianix, are developing compounds to enhance muscle strength. Research by investigators at University of Wisconsin - Madison, and supported by the Wisconsin Chapter of The ALS Association complement current clinical trials at Neuralstem. The investigators are focusing on the delivery of stem cells to muscles.
These projects underscore the goal of supporting a diverse yet strategic portfolio to ensure that new investigators and novel ideas are encouraged both in early discovery to better understand disease as well as testing approaches that may be more rapidly applied to clinical testing in patients.
Rosario Osta, Ph.D. University of Zaragoza, Spain
Skeletal Muscle microRNA and their target genes in mouse models of RNA
MicroRNAs (miRNA) are small regulatory molecules that bind specific cellular messenger RNAs (mRNA) and inhibit the translation of that mRNA to protein. In ALS mice expressing mutant SOD1, muscle enriched miRNA have been shown contribute to the maintenance of neuromuscular communication. However, it is unknown how miRNA are regulated in various stages of the ALS muscle pathology and in stem cells responsible for muscle regeneration. The investigators plan to characterize how miRNA are affected in most vulnerable and resistant muscles of mutant SOD1 mice, and subsequently test if the affected miRNAs change the behavior of cultured muscle stem cells. They aim to characterize target mRNA for these miRNA to uncover cellular processes affected in ALS muscle. Preliminary data indicates that miRNA in mutant SOD1 mice can be detected in circulating plasma, which may help to provide valuable biomarkers for detection of ALS. Based on data gained from the muscle tissue, the investigators plan to identify dysregulated miRNA in the blood of mutant SOD1 mice and test their applicability as biomarkers in human patient-derived blood samples. miRNA and their targets identified in the work will provide valuable information for development of diagnostic and therapeutic tools for ALS.
Gene Yeo, Ph.D. University of California, San Diego
Identification of an RNA signature of TDP-43 dependent misregulation in human neurons
A key breakthrough in understanding ALS pathogenesis came from the observation that an RNA binding protein, TDP-43, which is normally found in the nucleus misaccumulates in the cytoplasm of neuronal and glial cells in ALS patients. In 2006, mutations were found in TDP-43 accounting for 2-6% of familial cases, but it is increasingly evident that TDP-43 pathology affects sporadic ALS as well. The investigators have previously used cutting-edge genomic technologies and analyses to discover that TDP-43 binds directly to and regulates around ~1,000 genes in the brains of mice, many of which are crucial for the survival and proper function of mammalian neurons. In this project, investigators will confirm and extend this RNA signature in human neurons generated from human progenitor cells as a way to model ALS in a dish. Together with their published mouse data, this will define an evolutionarily conserved TDP-43 dependent RNA signature. They will develop this RNA signature into an in vitro assay to monitor the disease in a dish. The success of this project will greatly aid the future discovery of new compounds and treatments that will revert the aberrant disease-causing signature to a normal one.
Marcus Rattray, Ph.D. Reading School of Pharmacy, University of Reading, United Kingdom
Joe Sweeny, Ph.D. Department of Chemistry, University of Reading, United Kingdom
Synthesis and pharmacological characterization of novel riluzole derivatives
Currently, the only licensed therapy for the treatment of ALS is riluzole (Rilutek) which, on average, extends lifespan by several months. While riluzole is the best, and only neuroprotective compound in ALS, it is of relatively low efficacy. Better therapies are needed. The investigators intend to modify the chemical structure of riluzole, with the aim to produce a compound that has the beneficial properties of riluzole, but is predicted to be more effective in people living with ALS. We will use novel chemical techniques to generate hundreds of derivatives of riluzole. These compounds will be tested (screened) for their ability to protect motor neurons, and the best compounds modified again until the most potent are discovered. Since the pharmacological properties of riluzole are not fully understood, the investigators will use their novel riluzole derivatives to discover the exact way in which riluzole targets cells. This work is specifically aimed to generate a drug‐like molecule as a pathway to better therapeutic treatment of ALS.
Wilfried Rossoll, Ph.D. Emory University, Department of Cell Biology, Atlanta, GA
The role of TDP-43 in axonal mRNA trafficking and local translation in healthy and diseased motor neurons
(Supported by The Wisconsin Chapter of The ALS Association)
The TDP-43 protein has emerged as an important factor in the ALS disease process. The function of TDP-43 in motor neurons, the nerve cells controlling voluntary muscle movement that are specifically affected in ALS, is still poorly understood. While most efforts focus on the function of this protein in the cell nucleus, the investigators have found that it is also present in the long axonal processes that connect motor neuron cell bodies of the spinal cord with muscles throughout the body. As a novel approach, they propose to investigate the role of TDP-43 in axons and on how mutations in TDP-43 affect this function. Characterizing its function in axons is an important goal to better understand ALS disease mechanisms and identify novel therapeutic approaches.
Peter Carmeliet, Ph.D., M.D. Versalius Research Center, Leuven, Belgium
PHD 1 as a novel neuroprotective target in ALS
The pathogenesis of ALS involves oxidative stress, vascular hypoperfusion, metabolic imbalance, etc. The group of prolyl hydroxylase domain proteins (PHDs) is known to regulate each of these processes. Yet, barely anything is known about their role in the central nervous system or in neurodegenerative diseases. Because of the pleiotropic effects of these oxygen sensors, PHD inhibition carries the potential of targeting many pathogenetic mechanisms in ALS. The investigators preliminary results show indeed that SOD1 mice lacking PHD1 show reduced paralysis and prolonged survival. Therefore, this proposal is expected to yield valuable insights into how PHD1 can be used as novel drug targets to treat ALS and other motor neuron diseases. Given the initial data on a metabolic rewiring occurring in the PHD1 knock-out neuron, this project also holds the promise of unraveling novel exciting biological concepts of the relationship between metabolism and neuroprotection.
Udai Panday, Ph.D. LSU Health Sciences Center, New Orleans, LA
Molecular Basis of FUS/TLS-related ALS
The investigators have developed a novel fruit fly (Drosophila) model of ALS that recapitulates many key features of human FUS-related neurodegeneration. They will use their fly model for determining molecular mechanisms involved in causing ALS pathogenesis and investigating the impact of ALS causing mutations in the normal functions of FUS. They will also perform an unbiased genome wide scan to identify novel genetic modifiers of FUS-related ALS.
Haining Zhu, Ph.D. Dept. of Mol. & Cell. Biochemistry, University of Kentucky Research Foundation, Lexington, KY
Studying FUS-mediated Familial ALS using a Drosophila Model
Several amyotrophic lateral sclerosis (ALS) genes have been identified as their mutation can lead to familial ALS, including two genes encoding RNA processing proteins TDP-43 and fused in sarcoma (FUS). This project is focused on studying FUS since little is known about how FUS mutations cause motor neuron degeneration in ALS. The investigators have established transgenic Drosophila lines and the flies showed locomotion deficiency when FUS was expressed in motor neurons. They propose to characterize this model in detail to determine whether neuronal death or denervation at neuromuscular junctions is the primary event accounting for the impaired motor function in flies. They will use the in vivo model to address three questions regarding the etiology of FUS-mediated familial ALS. First, is subcellular localization of FUS critical to its toxicity and motor neuron dysfunction? Second, how does RNA binding affect the FUS toxicity in vivo? Lastly, do cell types other than motor neurons contribute to the disease pathogenesis or progression? The findings will likely provide critical insights into the mechanisms by which FUS mutations perturb the RNA processing pathways and ultimately lead to the disease. The knowledge generated in the proposed research will also provide much-needed future direction for developing ALS treatment.
David Morton, Ph.D. Department of Integrative Biosciences, Oregon Health & Science University of Portland, OR
Determining the functional contributions of TDP-43-regulated genes on Drosophila locomotion defects caused by TDP-43 dysfunction
A major impediment in developing therapeutic agents for the treatment of ALS is that little is known about the cellular and molecular mechanisms that underlie the disease. An important recent breakthrough was the identification of a protein named TDP-43 found in the cytoplasmic inclusions of motor neurons in post-mortem samples from ALS patients. It remains a critical task to identify the molecular pathways involved in not only the disregulation of TDP-43 but also its function in the normal healthy nervous system. TDP-43 is an RNA binding protein that recent research has suggested binds to and regulates the expression levels of a very large number (hundreds to thousands) of target genes. The next major task is to identify which of these genes are major players in the development of ALS. The primary goal of this proposal is to use the fruit fly, Drosophila melanogaster, as a model to rapidly identify which candidate target genes are directly involved in the cellular defects observed in both loss of function mutations of TDP-43 and over-expression of TDP-43, which serves as a model for TDP-43 proteinopathies.
Masatoshi Suzuki, Ph.D. University of Wisconsin - Madison, Madison, WI
Combining stem cell and growth factor delivery targeting to the skeletal muscle in ALS
(Supported by the Wisconsin Chapter of The ALS Association)
Investigators will use a rat amyotrophic lateral sclerosis (ALS) model to test the feasibility of stem cell delivery for treatment of ALS. Specifically, they will test the ability of progenitor cell grafts that produce and secrete growth factors to enhance growth factor delivery directly to skeletal muscles in SOD1G93A rats. Their proposal will determine whether a newly established line of skeletal muscle progenitor cells can enhance, or even possibly replace, established strategies of mesenchymal stem cell delivery in the skeletal muscles.
Howard Weiner, MD. And Oleg Butovksy, MD. Brigham and Women’s Hospital, Boston, MA.
Therapy of ALS based on peripheral Ly6Chi monocyte abnormalities in ALS
Supported by The Wisconsin Chapter of The ALS Association
Amyotrophic lateral sclerosis (ALS) is a fatal neurologic disease characterized by the degeneration of motor neurons. There is evidence that the immune system and inflammation play an important role in the disease process, though these mechanisms are not well understood. The investigators have initiated studies investigating the role of the immune system in the SOD 1 animal model of ALS and have obtained preliminary data, which supports a novel hypothesis of immune mechanisms in ALS. They found that there is progressive recruitment from the bloodstream of a white blood cell, called a monocyte, which enters the spinal cord and causes damage to the motor neuron. This is a pro-inflammatory cell. Using a novel antibody they generated they could downregulate the inflammatory properties of the monocyte and decrease the infiltration of these damaging monocytes into the spinal cord. Animals treated in this way survive longer and have less spinal cord damage. They propose to: 1) characterize these inflammatory monocytes in the mouse model; 2) investigate how the treatment leads to prolonged survival of the animals. 3) investigate both the blood and spinal fluid from patients with ALS to determine the degree to which these changes we observed can be seen in patients. This will serve as a basis both for disease monitoring and new approaches for therapy. Of note, the changes observed in animals occurred at very early stages of disease, even before the onset of clinical symptoms, which could be very helpful if applied to people at early stages of their illness.
Eric Schmidt, Ph.D. Rockefeller University, New York, NY
Cell type specific molecular profiling of corticospinal projection neurons during ALS progression
ALS is characterized by progressive paralysis and eventual mortality due to the degeneration and death of specific types of nerve cells in the brain and spinal cord that control muscle movements. The nervous system is made up of thousands of distinct cell types, making it difficult to identify and study the genes expressed uniquely in discreet kinds of cells. This is particularly true in the cerebral cortex where the ALS-vulnerable “upper” motor neurons reside. A novel technique developed in the investigator’ lab, called bacTRAP, utilizes transgenic mice that enable them to target a genetically defined population of nerve cells and then capture a “snapshot” of genes being expressed specifically in those cells at a given point in time. This study will use bacTRAP to identify genes that are regulated specifically in upper motor neurons at early and late stages of disease progression in the SOD1-G93A mouse model of familial ALS. The results obtained from this project will allow a better understanding of the molecular changes that occur in the vulnerable cells as they get sick and to identify novel candidate genes that may serve as therapeutic targets for early intervention.
Daryl Bosco, Ph.D. Univ. of Massachusetts Medical School, Worcester, MA
Investigating the Loss of Nuclear Function of ALS Mutant FUS/TLS
Cells are the functional unit of all living organisms. They are self-renewing, and go through a cycle of growth stages that prepare the cell to divide into two identical cells. This process is called the cell cycle and is strictly regulated in every cell type. Neurons are among the few cell types that are not self-renewing. Therefore, these cells have strict control over the cell cycle, keeping it suppressed at all times. When suppression is disrupted, the cell cycle is re-initiated, however this leads to cell death rather than cell division. Because neurons are not selfrenewing, their death has major consequences, leading to diseases such as ALS. Recently, mutations in the protein FUS/TLS were shown to cause familial forms of ALS. Accumulating evidence shows that FUS/TLS is normally involved in cell cycle suppression in other cell types, and thus ALS-linked mutations may potentially alter this function causing the death of motor neurons. Therefore, it is essential that we understand the role that FUS/TLS plays in suppressing the cell cycle in neurons, and whether ALS-linked mutations can abrogate this function.
Raymond Grill, Ph.D. University of Texas Health Sciences Center at Houston, Houston, TX
Targeting CNS inflammation in a mouse model of ALS with a dual inhibitor of COX and 5LOX
Riluzole is the sole FDA-approved treatment for ALS. However, Riluzole treatment enhances lifespan by only a few months. There is a clear need to identify novel interventions that are: 1) as, if not more, effective than Riluzole, and/or 2) able to enhance the efficacy of Riluzole when delivered in combination. Riluzole is a known substrate of the P-glycoprotein (Pgp), a component of the blood-brain-barrier responsible for restricting access to the central nervous system of a wide range of biological substrates. Pgp levels are known to be increased by inflammatory prostaglandins and leukotrienes when tested both in vivo and in vitro. Milane and colleagues recently demonstrated that Pgp expression is dramatically elevated and bioavailability of Riluzole diminished in a mouse model of ALS that exhibits high inflammatory activity. Licofelone is a next-generation anti-inflammatory drug that inhibits both cyclooxygenase and leukotrienes. Investigators will determine whether Licofelone treatment can suppress Pgp and both preserve function and enhance lifespan in the G93A mouse model of ALS. They will also determine whether Licofelone treatment with Riluzole enhances bioavailability of Riluzole and enhances efficacy compared to Riluzole treatment alone.
Gian Giacomo Consalez, Ph.D. Fondazione Centro San Raffaele de Monte Tabor, Milan, Italy
A transgenic mouse line reporting synaptic activity of cortical and spinal motor neurons: a tool to investigate early events of ALS.
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease affecting motoneurons, the nerve cells that control muscle activity. ALS neurodegeneration leads to the progressive loss of all voluntary movements, and eventually, to the inability for patients to breathe without assistance. The progression from disease onset to a patient's death takes an average of 2 to 5 years, a disease course that poses a heavy burden on patients and their families. To date, plenty remains to be learnt about the cause(s) and progression of ALS, a disease for which no effective cure is available. The development of novel tests to detect the onset of ALS and follow its clinical course would require the ability to look inside the spinal cord to detect the earliest signs of malfunction in its synaptic circuits. For these reasons, the investigators have developed an innovative genetic biosensor that they will test in mouse models of ALS. The technology developed in this project will allow scientists to monitor and visualize in real time the activity of synapses in the diseased nervous system. This novel approach will facilitate the analysis of early presymptomatic events that lead up to the full-blown clinical picture of ALS. Further, this mouse line will assist researchers in the development of effective biological and pharmacological treatments for disease prevention or containment.
The ALS Association is especially committed to bringing new concepts and methods into ALS research, and young scientists play an important role in this process. Funding is by the generosity of the Safenowitz family through the Greater New York Chapter of The ALS Association, in memory of Milton, who died in 1998 of the disease.
Shuying Sun, Ph.D. Ludwig Institute for Cancer Research, UCSD, San Diego, CA
Determining damage within specific cell types caused by SOD1, TDP43 and FUS/TLS mutations
Pathogenesis from the first genetic cause of ALS, dominant mutation in SOD1, is now proven to result from the damaged gene causing injury within both the motor neurons and the neighboring cells upon which they rely for support. However, what the damage is within each of these cell types is not established. With identification of causative mutations in TDP-43 and FUS/TLS in both familial and sporadic ALS, RNA processing alterations have been implicated in pathogenesis. It remains unknown in which cell type the mutant proteins produce the damage that drives either disease initiation or progression and the degree to which RNA metabolism is altered by disease causing mutants. The investigators propose to elucidate the contributions of damage within three nervous system cell types (motor neurons, astrocytes and oligodendrocytes) by identifying the global translational mRNA changes caused by SOD1, TDP-43, and FUS/TLS ALS-linked mutants within individual cell types, which are lineage labeled by an EGFP tagged ribosomal protein. They will also determine the potential convergent changes caused by the three different mutant genes, which may provide a test for whether most instances of ALS have common steps that could be targeted to slow disease. This work will lead to better understanding of disease mechanism, and provide rationale for therapeutic and diagnostic method development.
Amanda Phillips, Ph.D. Department of Neurology, Johns Hopkins University, Baltimore, MD
Focal transplantation of human neural progenitor cell derived astrocytes isolated from ALS patients into the rat spinal cord
Although ALS is a disease of motor neurons, multiple studies have shown that the non-neuronal, “glial” cells surrounding the motor neurons play an important role in disease. Specifically, the glial cells called “astrocytes” significantly influence the progression of paralytic disease in a mouse model of familial ALS. When the diseased astrocytes are replaced with healthy astrocytes in this model, the mice have increased survival. While these studies are intriguing, the role of astrocytes in human sporadic ALS which makes up 90% of ALS cases is still somewhat unclear. With their collaborators, the investigators have isolated astrocytes post-mortem from patients with both familial and sporadic ALS as well as from healthy controls to examine for potential differences between these diseased and non-diseased astrocytes. In order to study the astrocytes in the spinal cord environment, the investigators propose to inject these cells into the spinal cord of healthy rats and examine their interactions with motor neurons. Using this model, they will be able to determine whether the human astrocytes from familial or sporadic ALS patients damage healthy motor neurons in the rat spinal cord. The findings from this proposal may guide whether or not astrocyte targeted therapies should be developed for sporadic ALS.
Ole Wiskow, Ph.D. Harvard University Stem Cell and Regenerative Biology, Cambridge, MA
Phenotypic characterization of purified motor neurons from ALS patient specific induced pluripotent stem cells
Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease which is caused by the specific degeneration of motor neurons, responsible for all voluntary and involuntary movements. As it is very difficult to study the death of motor neurons in patients themselves we propose to create an ALS disease model in culture using human pluripotent stem cells. These stem cells have been derived by a reprogramming process from patient and healthy control individuals and can be directed to give rise to motor neurons in culture. In order to facilitate the identification of stem cell derived motor neurons in culture the investigators plan to introduce a motor neuron-specific genetic reporter into the stem cells. The particular advantage of such an optimized disease model system in culture is the accessibility of the affected motor neurons for a range of molecular biological analyses. Importantly, it is also possible to directly compare ALS-affected motor neurons with unaffected ones from healthy individuals. The goal is to identify so far unknown determinants responsible for ALS motor neuron degeneration. Knowledge of such key disease-related cellular pathways would allow for the generation of new drugs that specifically target these pathways and hopefully decrease and/or cease the motor neuron degeneration in ALS.
Mercedes Prudencio, Ph.D. Research at Mayo Clinic in Florida, Jacksonville, FL
Identification of Pathogenic TDP43 targets in vivo
TDP-43 has been recognized as the main component of ubiquitin-positive protein inclusions in ALS cases. Several recent studies demonstrate that TDP-43 truncation and aggregation are involved in TDP-43 neurotoxicity, and may play a role in ALS pathogenesis. TDP-43 aggregation is accompanied by its translocation into the cytosol, depleting it from the nucleus where it plays role in transcript regulation. The goal of this project is to identify key RNA targets regulated by TDP-43 that may be aberrantly altered in ALS. These studies will provide valuable insight regarding TDP-43 function and the mechanisms by which TDP-43 loss of function may contribute to neurodegeneration. Since the majority of ALS cases show TDP-43-associated pathology, the identification of RNA targets of TDP-43 misregulated in ALS tissue will likely lead to a better understanding of mechanisms that lead to neurodegeneration in ALS patients.
Ying Li, Ph.D. Johns Hopkins School of Medicine, Baltimore, MD
The role of human IPSC derived NG2 glia on human motor neuron survival: strong implication of NG2 glia in pathogenesis
Amyotrophic lateral sclerosis (ALS) is a progressive disease characterized by the loss of motor neurons with unknown reasons. Familial ALS accounts for about 10% of all ALS cases and the Zinc/Copper superoxide dismutase (SOD1) mutations are associated with 20% of familial cases. A4V is the most common SOD1 mutation in North America and correlates with short survival. Animal studies indicate that expression of mutant SOD1 in motor neurons determines disease onset, while expression of SOD1 mutations in astrocytes and microglia cells affects disease progression. Recently investigators discovered in ALS mouse models, 1) NG2 glial cells underwent dramatic changes, and 2) more importantly, deletion of mutant SOD1 in these NG2 glia significantly delayed disease onset, strongly suggesting that expression of mutant SOD1 in NG2 glia is neuropathogenic. In this project, investigators will characterize NG2 glia derived from ALS patient-specific induced pluripotent stem cells (ALS-iPSC) carrying the A4V mutation, and investigate the effects of these AV4-mutated NG2 glia on the survival of motor neurons equally derived from ALS-iPSC. This study will teach us whether mutant SOD1 has effects on human NG2 glia biology, and importantly, whether human NG2 glia cells play a role in ALS pathogenesis.
Frank S Walsh, Ph.D., Corey S Goodman, Ossianix Inc, University City Science Center, 3711 Market St, Philadelphia, PA 19104, USA
Reversal of muscle atrophy in amyotrophic lateral sclerosis by inhibition of myostatin via a single domain antibody (VNAR) therapeutic
ALS is characterized by death of motor neurons that leads to denervation of muscle fibres with a rapid development of muscle atrophy and loss of muscle function. A protein called myostatin has been isolated that serves to limit the amount of muscle tissue in the body. Neutralization of myostatin by an antibody has been found to dramatically increase the amount of muscle in the body and leads to increased strength. Investigators showed previously that in a mouse model of ALS that myostatin neutralization also built muscle even at advanced stages of the disease. Investigators will now develop a next generation therapeutic antibody using single domain antibodies that are more potent, smaller and act for longer than normal antibodies. This will be tested in animals and ultimately will be tested in patients with ALS for positive effects on muscle function particularly those for breathing and swallowing.