Completed Projects

Investigator:	Jeffrey A. Johnson, PhD
Organization:	University of Wisconsin, Madison
Project:	The Role of Oxidative Stress in Initiation and Progression of ALS in SOD G93A Mice
Funded:	3/01/04 - 2/29/08
Co-funded by ALSA
Defects in the mitochondria (energy producing machine of the cell) have been noted in both sporadic and SOD1-linked cases of ALS. Interestingly, a fraction of SOD1 localizes inside the mitochondria and this portion of SOD1 may contribute to motor neuron death in ALS. The goals of this study are to develop means for controlling the mitochondrial accumulation of SOD1 and for ultimately testing the role of mitochondrial SOD1 in ALS.
Investigator:	Elizabeth Fisher, PhD & Linda Greensmith, PhD
Organization:	Institute of Neurology, Queen Square, London
Project:	Investigating why dynein mutation ameliorates the SOD1 ALS phenotype and considerably extends lifespan
Funded:	2/01/05 - 1/31/08
Co-funded by ALSA
Mouse models are extremely helpful for investigating what goes wrong in a mid-life onset neurological disorder such as ALS. Mice give us access to tissues at all stages of development and disease progression, and because we share the same genes as mice and the same biochemical pathways (although not necessarily the same physiology), we can learn about the genetic causes of motor neuron degeneration from mouse strains with heritable forms of motor neuron disease. For some years, we have been working with the "Legs at odd angles" (Loa) mouse, which has a relatively mild but progressive form of motor neuron disease. We identified the mutated gene, a "dynein" gene, in 2001 (report published in the journal Science in 2003). We have now crossed this mouse to a widely-used mouse strain — called mutant SOD1 — that models human familial ALS. To our surprise, instead of the offspring having even greater neurological problems, the onset of their symptoms was considerably delayed. Further, their life span was extended by almost 34 percent, compared to their ALS model parents and siblings. We conclude there must be some kind of unexpected interaction between the Loa mutation and the SOD1 mutation. We hope to investigate the basic biology of the double mutant mice and collect information on their lifespan, cellular changes, muscle function, and transport processes within the neurons. We believe there is a good chance these surprising mice may shed light on what goes wrong in SOD1 related ALS, and tell us how to treat it.
Investigator:	Jean-Pierre Julien, PhD
Organization:	Laval University Research Centre
Project:	Development of Passive Immunization Approaches for ALS Caused by SOD1 Mutations (formerly Mouse Models to Study the Role of Microglia in the Pathogenesis of ALS)
Funded:	1/01/01 - 12/31/07
There is growing evidence that non-neuronal cells may contribute to ALS pathogenesis. This study proposes to determine to what extent microglia activation contribute to pathogenesis in mice models of ALS. The specific aims are: To generate ALS mice (SOD1G37R or SOD1G85R or SOD1G93A mice) expressing the HSV-thymidine kinase (tk) gene under the control of the myeloid CB11b gene promotor. Treatment with ganciclovir at different ages will be used to assess the contribution of microglia to disease. To determine if SOD1G37R - mediated disease is exacerbated in response to immunogenic challenges.
Investigator:	Nicholas Maragakis, MD & Mahendra Rao, MD, PhD
Organization:	Johns Hopkins University / National Institute on Aging
Project:	Mutant SOD1-derived Glial and Neuronal restricted precursors as tools for understanding cell autonomy in ALS
Funded:	8/01/04 - 7/31/07
Co-funded by ALSA
ALS (Lou Gehrig's Disease) is a disease of the brain and spinal cord in which the nerve cells that send signals to muscles slowly die. Patients with ALS become progressively weak and ultimately die of respiratory failure. ALS affects adults of all ages and there is no effective therapy for the disease. Why do motor neurons die in ALS? Many theories have been proposed but emerging studies suggest that it is not only disease within the motor neurons themselves but also other cells of the brain such as astrocytes, that contribute to the disease process. Astrocytes are responsible for maintaining the environment in which motor neurons function. Alterations in astrocytes may cause disrupt normal cell function and ultimately lead to motor neuron death. At some point, we need to understand the mechanisms underlying the relationship between astrocytes and motor neurons. To do that, we propose to isolate both astrocyte and neural stem cells from a mouse model of ALS. That gives us a suitable system to study their interactions in a tissue culture dish and should help shed light on pathways that lead to their malfunction in this disease.
Investigator:	Doug Kerr, MD, PhD
Organization:	Johns Hopkins University, Department of Pathology
Project:	Utilization of Embryonic Stem Cells in Motor Neuron Diseases
Funded:	8/01/04 - 7/31/07
Packard Center funding for this project was made possible by Regency Homes MDA Golf Classic. Co-funded by ALSA.
We have found that embryonic stem (ES) cells that have matured with characteristics of motor neurons survive when transplanted into adult rats with a spinal motor neuron injury. Many of the resulting cells resemble mature motor neurons. In this project, we will discover the best techniques to let us generate working ES-derived motor neurons in the injured adult mammalian spinal cord. One of our steps involves testing ways to block obstacles to the growth of newly-transplanted axons out of the spinal cord and into the peripheral nervous system. In another approach, we’ll test ways to enhance growth of the new motor neurons, allowing them to extend axons and form proper, functioning junctions with target muscles. In order to do that, we’ll provide natural agents that attract motor axons to target skeletal muscle. These studies look into transplantation as an important approach to motor neuron diseases such as ALS, SMA, and for spinal cord injury and other spinal disorders.
Investigator:	Richard Ransohoff, MD & Erik Pioro, MD, PhD
Organization:	Cleveland Clinic Foundation
Project:	The Role of Microglial CX3CR1 in the G93A SOD1 Transgenic Mouse Model of ALS
Funded:	7/01/05 - 6/30/07
Dr. Ransohoff is the Packard Center's inaugural Boye Foundation Researcher (2005)
Microglia are the very small, resident immune cells of the central nervous system (CNS). A body of research shows that they have two faces: They provide defense against bacteria and viruses, but their inflammatory properties endow them with the potential to injure delicate neurons. We discovered that neurons continuously manufacture and secrete a potent agent called CX3CL1 which apparently damps down damaging effects, acting directly toward CX3CR1, a receptor on microglia. Mice without the receptors or without the agent itself are susceptible to exaggerated forms of CNS damage in which their neurons and microglia die. Because inflammatory damage caused by microglia has a suggested role in ALS’s downhill process, we hope to explore this system thoroughly in animal models of the disease. We’ve performed one study in mouse ALS models—the standard mutant SOD1 mice—that also lack CX3CR1 receptors. Their loss of spinal cord motor neurons is severe; they live an even shorter time than typical SOD1 model mice. Now we’re exploring how activated receptors are able to restrain microglia. This work will help us gain insight into how the microglial reaction to initial neuronal injury in the disease can be either protective or harmful. Because existing anti-inflammatory medications can modify microglial behavior, our research may rapidly be translatable, in some way, as therapy.
Investigator:	Don Cleveland, PhD
Organization:	Ludwig Institute for Cancer Research, UCSD
Project:	Identifying how ALS2 contributes to motor neuron survival and function
Funded:	5/01/02 - 6/30/07
Investigator:	Mahendra Rao, MD, PhD & Tim Magnus, MD, PhD
Organization:	National Institute on Aging
Project:	Effects of Inflammation on Glutamate Transporters in Glial Restricted Precursors (formerly titled Generation of Human Astrocyte Precursor Lines)
Funded:	8/01/02 - 4/30/07
Abnormal glutamate metabolism may be the mechanism underlying motorneuron loss in ALS (reviewed in Maragakis and Rothstein 2001). Dr. Rao's laboratory has recently identified dividing glial precursor populations as well as cell surface markers that can be used to isolate these dividing cells. The ability to obtain purified dividing stem cells of the nervous system makes the process of cell line generation relatively straightforward. This project will generate astrocyte precursor cell lines that express the human glutamate transporter at high levels. The cell lines will be characterized in vivo and in vitro assays for their ability to protect motorneurons from glutamate toxicity.
Investigator:	Val Culotta, PhD
Organization:	Johns Hopkins University School of Public Health
Project:	The role of glutaredoxins in misfolding of ALS SOD1 mutants (formerly titled The impact of glutathione and the CCS copper chaperone on mutant polypeptides of SOD1)
Funded:	4/01/04 - 3/31/07
Funding for this project was made possible by MDA's Wings Over Wall Street®
Mutations in superoxide dismutase (SOD1) have been linked to inherited forms of ALS, although the underlying mechanism is poorly understood. We have identified two factors that interact with SOD1 in cells, and these include an abundant sulfur-containing molecule known as glutathione (GSH), and secondly, the copper chaperone for SOD1 (CCS) — that’s a molecule that normally carries copper needed for normal activity to the SOD1 enzyme. Our research tests the proposal that GSH and CCS have opposing effects on the mutant SOD1 polypeptide: while GSH might help the toxic process triggered by mutant SOD1 along, CCS may help protect against that toxicity.
Investigator:	Bryan Traynor, MD & John Hardy, PhD
Organization:	National Institute on Aging, NIH
Project:	Genome-wide association study of sporadic ALS using the 600K Illumina SNP chip set
Funded:	3/01/06 - 2/28/07
A fair amount is known about the key genes underlying ALS in the 10 percent of patients with the familial, or heritable, form of the disease. But the far more common sporadic form is a black box, genetically. No one knows, for example, if sporadic ALS is due to multiple gene interactions or to flaws in the way the environment affects certain genes. Such a lack of understanding delays an early diagnostic test for the disease. And it also slows the search for treatment. New Packard grantees John Hardy, Ph.D., and Bryan J. Traynor, M.D., with the National Institutes of Health, have begun a project that aims both to clarify the role of genes in sporadic ALS and to identify them. They’ll collect DNA from both ALS patients and healthy controls in the United States, and, using new, high-efficiency technology, will examine it for characteristic regions — called SNPs — associated with ALS patients but not with controls. Repeating the studies in matched Italian sporadic ALS patients will insure that any genetic differences found are ALS-related. Such association studies, as they’re called, greatly help pinpoint causative or susceptibility genes.
Investigator:	Shan Sockanathan, PhD
Organization:	Johns Hopkins University School of Medicine
Project:	Retinoid Responsive Genes in Motor Neuron Specification
Funded:	12/01/03 - 12/31/06
Funding for this project was made possible by MDA's Wings Over Wall Street®
Creating different specialized groups of motor neurons depends, in part, upon a key signaling molecule called retinoic acid (RA). We have identified a novel gene regulated by RA and have shown that it is essential for motor neuron generation. Our studies now focus on understanding how this gene works, something that may highlight a new signaling pathway critical for the development of motor neurons.
Investigator:	Kathryn Wagner, MD, PhD
Organization:	Johns Hopkins University School of Medicine
Project:	Myostatin Inhibition for Motor Neuron Disease
Funded:	12/01/04 - 11/30/06
Funding for this project was made possible by MDA's Wings Over Wall Street®
Part of the body’s internal system of checks and balances involves a protein called myostatin. It’s a self-made molecule that puts a brake on muscle growth. Research in mice, cattle and humans has shown that in the absence of myostatin, muscle growth approximately doubles. So far, scientists studying muscular dystrophy have used myostatin to great advantage in animal models of the disease. Now new Packard grantee Kathryn Wagner wants to see if blocking myostatin might help counter effects of motor neuron disease. She’s looking at the effects of loss of myostatin in the mutant SOD1 mouse model of ALS.
Investigator:	Moses V. Chao, PhD
Organization:	New York University
Project:	Mechanisms of Motor Neuron Apoptosis in ALS
Funded:	9/01/04 - 11/30/06
Amyotrophic Lateral Sclerosis (ALS) is a disease of the skeletal muscular motor neurons throughout the nervous system that results in the progressive loss of those motor neurons and muscular atrophy. Throughout development and in various neurodegenerative conditions, neurons die by a programmed cell death mechanism, termed apoptosis, carried out by one or more known pathways. Moses Chao is studying specific pathways leading to neuronal death to gain insight into the underlying causative agent of ALS. He hopes that will let him devise strategies for early diagnosis as well as therapeutic intervention.
Investigator:	Wim Robberecht, MD, PhD
Organization:	University of Leuven, Belgium
Project:	A transgenic zebrafish model to study the pathogenesis and treatment of ALS
Funded:	10/01/04 - 9/30/06
To help make sense of the many leads that exist on what causes ALS or on how it develops, scientists need more animal models than the classic SOD1 mouse. This is especially true for sporadic ALS, in which several factors likely work together to start the disease. In the present project, Belgian researcher Wim Robberecht intends to develop a model for ALS using the zebrafish. Imbuing fish with human genes—making transgenic animals—is much easier than using rodents. Also, the faster reproductive rate of fish and large number of progeny will make many more transgenic animals available. It’ll enable studies to be completed more quickly. Also, larger numbers make results more reliable. With such a model, Robberecht’s group will go on to study the effects of genes thought to be important in ALS, either as possible causes of the disease or as agents that shape the way the disease unfolds.
Investigator:	Ruben J. Boado, PhD
Organization:	ArmaGen Technologies / UCLA
Project:	Genetic Engineering of a Recombinant Neuroprotective Neurotrophin that Crosses the Blood-Brain Barrier
Funded:	9/01/05 - 8/31/06
Co-funded by the Brain Trust Collaborative
A new therapy for chronic neuro-degenerative diseases such as Alzheimer's or Parkinson's disease or ALS may be developed by genetically engineering two unrelated genes to combine and produce a new protein. The result, a protein known as AGT-140, contains a portion that enables it to cross the human blood-brain barrier (BBB)—a natural wall between the circulatory system and the brain designed to exclude certain molecules. AGT-140 also has regions that encourage neuron survival. The protein ingeniously taps into a natural mechanism cells use to carry insulin across the BBB. Once inside the adult central nervous system, says researcher Ruben Boado, it should target a receptor that unleashes chemical cascades protective of neurons or that encourage their growth. In short, the novel approach makes central nervous system pharmaceuticals out of naturally occurring neurotrophins.
Investigator:	Kati Andreasson, MD
Organization:	Johns Hopkins University School of Medicine
Project:	Function of Prostaglandin Receptors in Models of ALS
Funded:	12/01/03 - 8/15/06
Funding for this project was made possible by MDA's Regency Homes Golf Classic
In initial studies, we have discovered that a class of signaling molecules, the prostaglandins, has a profound impact on the survival of motor neurons in the spinal cord. We have identified an initial group of prostaglandin receptors which, when activated by specific drugs, can rescue motor neurons in a spinal cord model of ALS from the toxic effects of too much of the neurotransmitter glutamate. Our findings thus far suggest a novel therapeutic strategy in ALS that targets specific pro-survival prostaglandin receptors. We propose in this grant application to study other prostaglandin receptors, in hope of identifying ones that promote neuronal survival and those that may promote toxicity. These studies will lay the groundwork for studies on the usefulness of prostaglandin receptor-binding drugs in models of ALS.
Investigator:	Craig L. Bennett, PhD
Organization:	University of Washington
Project:	Mutations in the ALS4 gene (SETX) may alter RNA processing in motor neurons
Funded:	8/01/04 - 7/31/06
Packard Center scientist Craig Bennett has recently uncovered mutations in the Senataxin gene (SETX) as the molecular basis for ALS4, a rare form of juvenile onset amyotrophic lateral sclerosis (ALS). His team and others have identified a specific enzyme portion of the protein encoded by the ALS4 gene that helps manipulate DNA. The researchers believe it may provide clues to the gene's normal purpose and to the molecular pathways that may go awry in ALS4. Bennett's team has conducted experiments to (1) verify that the ALS4 protein does indeed have possess enzyme activity and see if the observed mutations affect it (2) to test if mutations of the ALS4 protein outwit an internal policing system cells have to keep mutant proteins from appearing and (3) to test if the mutations affect the cell's ability to mop up an excess of the chemical glutamate, which is toxic in high amounts.
Investigator:	Daniel Drachman, MD
Organization:	Johns Hopkins University School of Medicine
Project:	Treatment of ALS by gene transfer of baculoviral p35 to block apoptosis of motor neurons
Funded:	7/01/04 - 6/30/06
Funding for this project was made possible by MDA's Wings Over Wall Street®
The overall goal of this project is to protect motor neurons from death in a mouse model of ALS. Although many different factors may contribute to ALS, the actual loss of nerve cells takes place by a process of apoptosis. This involves triggering a cascade of enzymes (caspases) that lead to motor neuron death. The caspases can be powerfully blocked by a protein substance called baculoviral p35. We are developing a system to deliver the protein-producing p35 gene to neurons using a non-harmful viruses. We have already shown, in neuron cultures, that p35 delivered by an adenoviral vector prevents damage of human brain cells subjected to agents that cause apoptosis, including tumor necrosis factor alpha, and the HIV proteins gp120 and Tat. We have also shown that adenoviral vectors, when injected into muscles of intact mice, are carried back into motor nerve cells, where the target genes they carry are expressed. In this project, we will use viral vectors (adenovirus and adeno-associated virus) to deliver the p35 gene to motor neurons in mouse models of ALS. Then we’ll evaluate the beneficial effects of this treatment.
Investigator:	David R. Borchelt, PhD
Organization:	Johns Hopkins University / University of Florida
Project:	Role of metal cofactors in the toxicity of FALS mutant SOD1
Funded:	1/01/02 - 6/30/06
Funding for this project was made possible by MDA's Wings Over Wall Street®
Mutations in Cu/Zn superoxide dismutase 1 (SOD1) have been linked to dominantly inherited forms of amyotrophic lateral sclerosis (FALS). Three prevailing hypotheses have been postulated to explain the toxicity of mutant SOD1: enhanced peroxidase activity, enhanced nitration of tyrosines, and the formation of aggregates. Consequently, this research will focus on three specific projects: (1) Defining the role of copper-mediated chemistry in the toxicity of mutant SOD1; (2) characterization of protein aggregates in the spinal cords of SOD1 transgenic mice and patients with ALS; and (3) generation of transgenic mice expressing mutant SOD1 predominantly in neurons, astrocytes, and muscle.
Investigator:	John Gearhart, PhD
Organization:	Johns Hopkins University, Institute for Cell Engineering
Project:	Directed Differentiation of Human Pluripotent Stem Cells in Motor Neurons
Funded:	7/01/03 - 6/30/06
Funding for this project was made possible by MDA's Wings Over Wall Street®
Pluripotent stem cells may be differentiated into specific lineages for eventual use in transplant-based therapies. We propose expanding upon our previous success with directing mouse embryonic stem cells into motor neurons to include human pluripotent stem cells. Several growth factors including sonic hedgehog, bone morphogenetic proteins, and fibroblast growth factors will be used to direct differentiation into specific motor neuron lineages. We will purify differentiated motor neurons and examine their function in-vitro.
Investigator:	Marie Filbin, PhD
Organization:	Hunter College, City University of New York
Project:	Strategies to encourage grafted ES-derived motor neurons to regenerate in vivo
Funded:	11/01/03 - 1/31/06
Recently, a great effort by a handful of scientists has made it possible to produce large numbers of motor neurons from embryonic stem cells. Unfortunately, when these motor neurons are transplanted into animals they do not re-grow, largely because of inhibitors in the environment. Marie Filbin and her laboratory team have devised ways to reverse this damping down by chemically overriding a motor neuron's response to inhibitors. They're presently testing these strategies for the motor neurons made from embryonic stem cells.
Investigator:	David Borchelt, PhD
Organization:	Johns Hopkins University School of Medicine / University of Florida
Project:	Mapping the Toxic Domains of SOD1
Funded:	2/01/03 - 1/31/06
Funding for this project was made possible by MDA's Wings Over Wall Street®. Co-funded by ALSA.
Some inherited forms of ALS are caused by mutations in a protein called superoxide dismutase 1 (SOD1). Recent research suggests the mutations cause the protein to acquire some type of toxic property. Some of the disease causing mutations in SOD1 abnormally shorten the protein, deleting some of the amino acids at the end of the chain. David Borchelt wonders how the shortening of the protein leads to its toxic behavior and its ability to trigger inherited forms of ALS. He aims to introduce, into mice, human SOD1 genes with mutations that progressively shorten the subsequent protein it codes for—to identify the least length it takes to cause disease. He will also methodically knock out internal parts of the SOD1 protein to pinpoint which parts are responsible for its toxic effects.
Investigator:	Richard L. Huganir, PhD & Rita Sattler, PhD
Organization:	Johns Hopkins University, Department of Neuroscience
Project:	Role of AMPA receptor subunits and interacting proteins in excitotoxic processes of ALS
Funded:	8/01/04 - 7/31/05
Funding for this project was made possible by MDA's Wings Over Wall Street®
Glutamate is the most common excitatory neurotransmitter in the central nervous system (CNS) and activating its corres-ponding receptors enables rapid flow of nerve impulses in the mammalian brain. While learning and memory, as well as other plastic changes in the CNS involve glutamate receptor activity, excessive stimulation of these receptors is thought to underlie a number of CNS diseases including ischemia, trauma and neuro-degenerative diseases such as Huntington's disease, Alzheimer's disease and amyotrophic lateral sclerosis (ALS). Our research aims to explain the workings of the AMPA type of glutamate receptor and its influence throughout CNS neurons, both under normal conditions and in disease.
Investigator:	Ron Oppenheim, PhD
Organization:	Wake Forest University, Departments of Neurobiology and Anatomy
Project::	The Effect of Bax Deletion on Motor Dysfunction and Neurodegeneration in the SOD1 G93A Mutant Mouse
Funded:	12/01/02 - 7/01/05
When ALS kills motor nerves, it unleashes a process that ultimately trips a destructive program hard-wired into motor neuron cells. Programmed cell death has been eyed as a possible target for therapy for ALS and other neurodegenerative diseases, and scientists have singled out certain steps on that downward path as being potentially more useful than others to disrupt. Neuroscientist Ronald W. Oppenheim is investigating a possible target in a gene called Bax, a key player in the self-destruct process. Oppenheim's found that mice lacking Bax resist a natural clearing of unnecessary neurons that comes with normal embryo development. Unfortunately, however, the rescued nerve cells fail to connect properly with target muscles or to integrate properly into spinal cord networks. But Oppenheim has evidence that mice injected with the substance GDNF (glial derived neurotrophic factor) have more normal-appearing nerve cells. Also, many more motor neurons show proper muscle connections. In his Center work, Oppenheim will study mice without the Bax gene, only this time in mouse SOD1 models of ALS. He'll monitor neuromuscular function and motor nerve behavior in the mice to see if lacking Bax protects the mice, if it's in some way harmful or if ALS-like changes appear. Should nerve cells be protected from death but fail to connect with muscle, he'll then test GDNF's ability to correct that and do followup studies to explain precisely what's happening.
Investigator:	Doug Kerr, MD, PhD
Organization:	Johns Hopkins University, Department of Pathology
Project::	Motoneuron-differentiated ES cells in ALS
Funded:	7/01/03 - 6/30/05
Funding for this project was made possible by MDA's Wings Over Wall Street®
This project will utilize mouse embryonic stem (ES) cells to define critical molecular cues leading to motoneuron death in ALS and to explore their potential for restoring function in ALS. Embryonic stem cells from mutant SOD1 mice will be differentiated into motoneurons and grown in co-culture with skeletal muscle. We will examine when in the process of differentiation these cells become abnormal and will characterize the early alterations in cellular function that ultimately result in motoneuron death. In a parallel project, we will transplant normal ES cells that have been differentiated into motoneuron progenitors into SOD1 mice and rats. These cells will be examined for their ability to survive as motoneurons within the SOD1 spinal cord and to extend axons through surrounding white matter.
Investigator:	Clarke Tankersley, PhD
Organization:	Johns Hopkins University School of Public Health
Project::	Reversing Respiratory Insufficiency in SOD1G93A Mice
Funded:	4/01/03 - 3/31/05
Funding for this project was made possible by MDA's Wings Over Wall Street®
In ALS, the decline in breathing comes from the gradual death of motor neurons that serve the muscles of the diaphragm and accessory muscles in the upper body (the intercostals). As nerve stimulation ebbs, these target muscles atrophy. Because loss of breathing is the major cause of death in the disease, helping restore those muscles and their nerve input is the goal of our work. Meanwhile, we're not averse to a way to greatly slow the decline in breathing and so extend good quality of life. Using viruses, we've developed a way to get possibly therapeutic genes into respiratory motor neurons in model mice (those carrying a human SOD1 mutation). Such genes could code for proteins that extend a nerve cell's ability to resist damage or to repair itself. But before we undertake animal trials, we need to understand exactly how breathing changes in the model mice, how the pattern of breathing is altered and to what extent. That will allow us to track the progress of the therapies we try.
Investigator:	Philip Wong, PhD & Huaibin Cai, PhD
Organization:	Johns Hopkins University School of Medicine; National Institute on Aging
Project::	Development of a Mouse Model of Amyotrophic Lateral Sclerosis 2 (ALS2)
Funded:	2/1/03 - 1/31/05
Packard Center funding for this project was made possible by MDA's Wings Over Wall Street®. Co-funded by ALSA.
A new gene called ALS2, that is linked to an autosomal recessive juvenile form of ALS (Amyotrophic Lateral Sclerosis) has recently been identified. Unlike classical ALS, the onset of juvenile ALS is usually before the age of 25 and disease progression is more protracted. We plan to use a genetic approach to create a mouse model of juvenile ALS that may help us to understand the cause of this form of ALS and provide clues as to how ALS2 is related to the SOD1-linked cases of ALS. Importantly, our studies may also have implication for therapeutics and may shed light on how motor neuron loss occurs in the more general form of sporadic ALS.
Investigator:	Don W. Cleveland, PhD and Larry Goldstein, PhD
Organization:	Ludwig Institute for Cancer Research, University of California in San Diego; Howard Hughes Medical Institute, UCSD
Project::	Determination of which cells are directly damaged by familial ALS-linked SOD1 mutants
Funded:	9/01/02 - 8/31/04
Goldstein and Cleveland build on their previous Center work to see if ALS begins exclusively in damaged motor neurons. The researchers studied chimeric mice—those comprised of normal cells plus cells carrying the same SOD1 mutation that causes an inherited form of the disease in humans. This mix of cells, where normal ones are in contact with disease prone ALS mimics, has shown that the toxic process of the disease doesn't originate in motor nerve cells alone. From their results, it's possible, say Goldstein and Cleveland, that none of the crucial toxicity starts there. In this new study, the researchers hope to find which cells are the first targets of the destructive process that mutant SOD1 triggers. They’ll do this by selectively switching off expression of the damaging SOD1 mutation in key cells: neurons, glia, microglia or muscle cells. The study also aims to show how SOD1-caused ALS progresses and to estimate the time required for each stage. The researchers will also see if eliminating the damage-causing protein can slow or stop disease progression. Such work is important to understanding the disease, as well as a preparation for any sort of replacement therapy, including that of stem cells.
Investigator:	Richard O'Brien, MD, PhD
Organization:	Johns Hopkins University School of Medicine
Project::	The Regulation of Glutamate Receptor Expression in Spinal Motorneurons
Funded:	8/01/00 - 7/31/04
The goal of this research is to understand the molecular controls on AMPA type glutamate receptors in motor neurons as a first step towards rational interventional pharmacotherapy. Specifically, investigators are determining how the formation of excitatory synapses on motor neurons modulates the number and distribution of glutamate receptors through: Isolating and characterizing molecules involved in synapse formation Determining how the dysfunction could lead to disease states Examining the molecules involved in the synaptic targeting of glutamate receptors in motoneurons Examining the role of the glutamate receptor subunit GluR2 in motor neurons' survival in vitro
Investigator:	Vassilis E. Koliatsos, MD
Organization:	Johns Hopkins University School of Medicine
Project::	Neural Stem Cell Therapies of Experimental Models of Motor Neuron Disease
Funded:	8/01/00 - 7/31/04
The purpose of this study is to replace lower motor neuron populations by Neural Stem cells in order to restore structure and function first in animal models and then in patients with ALS.
Investigator:	Alex Kolodkin, PhD
Organization:	Johns Hopkins University School of Medicine
Project::	Neuropilin and Class 3 semaphorin function in motor neurons
Funded:	9/01/00 - 7/31/04
The aim of this study is to understand how damaged neurons might be stimulated to regenerate and extend processes back to their targets. The researchers use a mouse model to investigate the roles played by certain cellular factors that may contribute to motor axon degeneration. The study investigates how neurons that are susceptible to degeneration can be protected from damaging factors in their cellular environment.
Investigator:	Val Culotta, PhD
Organization:	Johns Hopkins University School of Public Health
Project::	Mitochondrial SOD1 and ALS
Funded:	4/01/02 - 3/31/04
Defects in the mitochondria (energy producing machine of the cell) have been noted in both sporadic and SOD1-linked cases of ALS. Interestingly, a fraction of SOD1 localizes inside the mitochondria and this portion of SOD1 may contribute to motor neuron death in ALS. The goals of this study are to develop means for controlling the mitochondrial accumulation of SOD1 and for ultimately testing the role of mitochondrial SOD1 in ALS.
Investigator:	Richard L. Huganir, PhD
Organization:	Johns Hopkins University, Department of Neuroscience
Project::	Role of AMPA Receptor Modulation in ALS
Funded:	9/01/00 - 1/31/04
The effective use of anti-glutamatergic drugs in recent clinical trials with ALS patients has supported the hypothesis that excitotoxicity mediates the motor neuron degeneration that occurs during ALS. In this study investigators are examining the role of the modulation of AMPA receptor by phosphorylation and GRIP on excitotoxicity in cortical, and spinal cord neurons and determining the possible role of AMPA receptor modulation in ALS. The results from this research may address basic mechanisms in excitotoxicity in ALS and other acute and chronic neurodegenerative disorders.
Investigator:	Jeffrey Elliott, MD
Organization:	University of Texas, Southwestern Medical Center
Project::	Mutant SOD1 in ALS Structure/Function Analysis
Funded:	10/01/01 - 12/31/03
Although changes in glia accompany the neuronal loss observed in a murine model of FALS, the significance of non-neuronal cell contribution to the disease process remains unclear. To address this question, this project seeks to alter expression of proteins found either in neurons or in glia and then ask whether such manipulations, independently, could influence the disease phenotype. It is hypothesized that these novel SOD1 proteins containing multiple mutations will exhibit altered catalytic function or aggregation potential compared to disease producing SOD1 molecules with a single mutation.
Investigator:	Paul F. Worley, MD
Organization:	Johns Hopkins University, Department of Neurology and Neurosciences
Project::	The Role of NARP (Neuronal Activity Regulated Pentraxin) in the Pathogenesis of ALS
Funded:	8/01/00 - 11/30/03
Investigators in this study are assessing the contribution of NARP (Neuronal Activity Regulated Pentraxin) in ALS pathogenesis by generating mice that lack NARP and express a pathogenic form of SOD1 and using behavioral studies to measure the rate of progression of weakness and motor control. The theory is that if NARP is involved in the adaptation process, the rate of progression of disease should be accelerated.
Investigator:	Dwight E. Bergles, PhD
Organization:	Johns Hopkins University, Department of Neuroscience
Project::	GLT-1 Transporter Action at Excitatory Synapses in ALS
Funded:	9/01/01 - 8/31/03
The mechanisms responsible for causing the degeneration of motor neurons in ALS are not known, however, research conducted over the last ten years points towards an essential role of the EAAT2 glutamate transporter in the etiology of this disease. Based on the hypothesis that increasing either the number of GLT-1 transporters, or the activity of existing GLT-1 transporters, will be neuroprotective and slow down the progression of motor neuron death in ALS, this project has three goals: (1) to develop new methodologies for monitoring GLT-1 activity in brain slices, (2) to determine whether GLT-1 transporters influence the activation of receptors during synaptic activity, and (3) to evaluate whether glutamate transport into astrocytes surrounding spinal motor neurons is compromised in SOD1 mutant mice.
Investigator:	David Irani, MD
Organization:	Johns Hopkins University, Department of Neurology
Project::	Virus-Induced Excitotoxic Motor Neuron Injury
Funded:	4/01/02 - 3/31/03
This laboratory group studies Sindbis virus (SV) infection in mice, a pathogen that targets motor neurons. A neuron adapted viral strain (NMSV) can cause lumbar motor neuron destruction with accompanying hindlimb paralysis. Our aim is to develop an animal model where acute excitotoxic motor neuron injury can be conveniently studied.
Investigator:	Gregory A. Cox, PhD
Organization:	Jackson Laboratory
Project::	Mapping the Hereditary Canine Spinal Muscular Atrophy Gene
Funded:	1/01/01 - 12/31/02
Hereditary Canine Spinal Muscular Atrophy (HCSMA) is a semi-dominantly inherited disorder of lower motor neurons that produces weakness, muscle atrophy, and paralysis. To understand the molecular events leading to motor neuron disease in the HCSMA dogs, we need to first know the nature and pattern of expression of the gene product. Thus we propose to genetically map the canine HCSMA locus and identify positional candidates by cross-species comparative genomics in the human and mouse.
Investigator:	Valina Dawson, PhD
Organization:	Johns Hopkins University, Department of Neurology
Project::	Mitochondrial Endonuclease G in Neurotoxicity
Funded:	12/01/01 - 11/30/02
Mitochondria play a key role in the initiation and amplification of cell death signals in ways that exceed simply providing for cellular bioenergetics. Additionally, mitochondria play a key part in the regulation of cell death through the release of proteins that are involved in cell death programs. Recently a new protein, mitochondrial endonuclease G (endoG), was discovered. This research project will investigate the role of endoG in neurotoxicity by generating antibodies to murine endoG and generating an endoG null mouse. These antibodies and this mouse will be used to investigate excitotoxicity. The endoG null mice will also be crossbred to FALS mice.
Investigator:	Don W. Cleveland, PhD and Larry Goldstein, PhD
Organization:	Ludwig Institute for Cancer Research, University of California in San Diego; Howard Hughes Medical Institute, UCSD
Project::	To Determine Whether Toxicity of Familial ALS-Linked SOD1 Mutants is Cell Autonomous
Funded:	9/01/00 - 8/31/02
There is a possibility that primary toxicity may be derived not simply through motor neurons, but also may be interpreted as damage accruing within astrocytes. The purpose of this research is to test in which cells the primary toxicity of SOD1 mutants is generated via: Constructing and analyzing new sets of transgenic animals bearing newly designed gene constructs for yielding either neuronal or glial specific expression Generating chimeric mice comprised of a combination of normal cells and cells expressing an SOD1 mutant
Investigator:	Margaret Sutherland, PhD
Organization:	George Washington University
Project::	Over-expression of EAAT2 in ALS
Funded:	9/01/00 - 8/31/02
We have developed a transgenic mouse that over-expresses the dominant astroglial glutamate transporter EAAT2. We will determine if excess EAAT2 protein can delay disease onset and prolong survival in the ALS mouse model, G93A SOD1. In our preliminary studies we have observed very significant slowing of disease progression, with survival in mice as long as 1 year longer than the usual death of the mice at 4 months old.
Investigator:	Russell Margolis, MD
Organization:	Johns Hopkins University, Department of Psychiatry
Project::	Study of a Large Family with Autosomal Dominant Spastic Paraplegia
Funded:	11/01/00 - 10/31/01
Spastic Paraplegia is a progressive neurodegenerative disorder primarily affecting the lower limbs with some similarities to ALS. We have recently identified a large family with a hereditary form of this disorder. We plan to begin genetic studies with the goal of eventually finding the genetic mutation that causes the neurological disease in this family. The discovery of this mutation may provide additional insight into the causes of ALS and other neurodegenerative disorders.
Investigator:	Valina Dawson, PhD
Organization:	Johns Hopkins University, Department of Neuroscience
Project::	The Role of Apoptosis Inducing Factor (AIF) in ALS
Funded:	9/01/00 - 8/31/01
Researchers for this grant hypothesize that Apoptosis Inducing Factor (AIF) plays a key role in excitotoxic neuronal death. Dr. Dawson proposes that mitochondria are stimulated to release AIF in stressed and injured cells, which may be the irreversible execution phase of excitotoxic cell death. The investigators, therefore, are studying the role of AIF in motor neuron injury and Familial ALS first by generating an AIF null mouse and then using the mouse to investigate excitotoxicity in motor neuron cultures.
Investigator:	Philip Wong, PhD
Organization:	Johns Hopkins University, Department of Pathology
Project::	The Development of a Conditional Model of Motor Neuron Disease
Funded:	8/01/00 - 7/31/01
Researchers in this study are generating and characterizing conditional transgenic models using the tetracycline-responsive gene system to control the expression of mutant SOD1 and to assess the degree and stage of disease reversibility. The conclusions will provide information regarding disease mechanisms, therapeutic approaches and optimal time windows for treatment.