Dystonia International Patient Registry (DIPR)    The Dystonia International Patient Registry (DIPR) is an observational registry of individuals who carry the DYT-1 gene.  The ultimate goal of this registry is to provide  the natural history of DYT-1 Dystonia worldwide.  All scientific and medical investigators are encouraged to utilize the registry as a source of patients for clinical research and as a source of demographic information on the patient population. Click here to access the registry.

 2011 Projects funded:

Patient-specific induced pluripotent stem (iPS) cells as a model for DYT1 dystonia

Nutan Sharma, MD PhD and Cristopher Bragg, PhD

Massachusetts General Hospital/Harvard University

The objective of this research is to establish a collection of DYT1-specific induced pluripotent stem (iPS) cells, which are stem cells derived from patient-donated somatic (skin) cells, and matched controls that will serve as a public resource for the dystonia research community. These cells, which are generated by reprogramming skin fibroblasts resemble embryonic stem cells in culture and can be differentiated into specific cell types, such as neurons, needed for research. Thus, DYT1-specific iPS cells will represent a self-renewing population of cells bearing ‘natural’ levels of the mutated target protein, torsinAΔE, capable of becoming human neurons, which are thought to be critical in developing DYT1 dystonia. This model would enable multiple dystonia research groups to conduct new studies designed to study neuron-specific defects associated with torsinAΔE.

The current plan is to perform an initial analysis of DYT1 vs. control-derived neurons and publish the findings, at which point the cells will be deposited with the Coriell Institute for public distribution. The Institute has already been notified about the intent to deposit the DYT1 iPS cells.

Project FireSky

BioFocus

BioFocus target discovery platform uses human cells to model diseases in combination with functional genomics. BioFocus introduces fragments of genetic material, DNA or RNA into cells using collections of specially designed viruses to study the roles of individual genes in disease processes. Advanced readouts are then used to identify the genes that have potential effect on the disease-related process in this model system. After further validation assays, these targets form the basis for the development of new drugs. This unique approach combines cell assays with high-throughput technology, supported by state-of-the-art monitoring equipment.

Outline of the Project so far

·        The major objective of the Project is to identify genes and proteins that modify the DYT1 dystonia phenotype.

·        To accomplish that a genetic approach is used. It is based on silencing (“turning off”) selected genes by using small RNA molecule specific for individual genes.

·        The RNA molecules target more than 4,500 genes preselected by BioFocus as the most promising, drugable targets.

·        During silencing torsinA function is monitored in cultured cell using a quantitative detection assay developed in Phase 1 of the Project.

·        Several rounds of robotic screening and selection of specifically interfering genes and proteins have been performed in Phase 2.

·        Identified genes and proteins will be potential drug targets for DYT1 and possibly other dystonias.

·        Selected targets will be utilized in subsequent drug discovery programs to develop therapeutics to modify the phenotype of DYT1 dystonia.

Phase 1 – Assay Development - completed

Phase 2 – Screening - ongoing

An assay using the neuron-like cell line to measure the secretion of proteins upon viral ‘suppression’ of torsinA was developed during Phase 1, and it is this assay that is being used during Phase 2 to screen the BioFocus’ SilenceSelect™ library. The SilenceSelect™ library targets the human drugable genome (>4,500 genes that are targeted by 12,000 viruses), which ensures that the identified targets are tractable for small molecule drug discovery.  The screening is done from an independent re-propagation of the viral stocks to confirm activity. Re-screening will reduce the number of hits eliminating some that are produced due to the experimental procedure or secondary effects. Additional toxicity experiments will also be performed to appropriately adjust the screening conditions.

Phase 3 – Target validation

In the last phase of the project, a narrowed down selection of validated target will be screened again by using a different assay and cell biological methods. The goal is to identify a manageable number of validated target that are likely players in the disease process. When successfully completed this will allow for identification of genes and proteins affecting torsinA function that might become targets for rational drug design. The identified targets will also enrich our knowledge about the role of torsinA in neurons and guide future research in this area.

2010 Summit - Dr. Edward V. Staab Memorial Grants

Nicole Calakos, M.D., Ph.D., Duke University Medical Center

Title: Novel high-throughput approach to screening for modifiers of TorsinA pathology

Hypothesis: The identification of modifiers of cellular inclusion pathology caused by mutant TorsinA proteins will provide novel targets to consider for the treatment of DYT1 dystonia.

Dr. Calakos’ group is developing methodology to perform high-throughput screens for modifiers of TorsinA dysfunction. Numerous assays have shown that disease-associated mutant forms of TorsinA behave differently from the normal protein. The Calakos group proposes to use the abnormal characteristics of disease-associated TorsinA to develop a high-throughput assay with high-content readouts to screen for modifiers that will reduce the abnormal protein burden.  The aim of this work will be to identify new candidate small molecules and pathways for the correction of pathological TorsinA activity. The ultimate goal of this work is to provide new opportunities for the treatment and cure of dystonia.

Dr. Ramon Rodriguez UFMDC

Ampicillan Trial

Dystonia is a disorder characterized by abnormal postures resulting from involuntary muscle contractions. It can be caused by a variety of conditions, including genetic factors. A mutation in a gene called DYT-1 gene was identified as an important contributor for early onset generalized dystonia. This gene produces an abnormal protein called torsinA, believed to play an important role in the abnormal muscle contraction causing dystonia. Although there are treatments for the symptoms of dystonia, at this moment there is no cure. Recently, a study found that by treating a mouse with a similar DYT-1 mutation with ampicillin (an available antibiotic), there were improvements in the mouse’s ability to walk. We will conduct a study to look at safety and tolerability of ampicillin.

At the University of Florida’s MDC, we  will conduct a study to look at safety and tolerability of ampicillin to improve dystonia symptoms under the clinical trial, “Ampicillin for DYT-1:a Pilot Trial for Safety and Efficacy.”   During the clinical trial patients will receive either study drug (ampicillin) or a placebo for a period of four (4) weeks. Then, there will be one (1) week ‘washout period’ with no study-related medications, followed by a switch to either the placebo or ampicillin for another period of four (4) weeks. As a double-blinded study, neither patient nor the investigator will know which pill patients are taking at any given time. Patients will come to our center five (5) times for evaluation of how they are tolerating the medication(s) and evaluation of their dystonia symptoms.

D. Cristopher Bragg, Ph.D.

Massachusetts General Hospital

Harvard Medical School

 “Accelerating drug discovery for dystonia using chemical genomics” which has been supported in the past year by the Dr. Edward V. Staab Memorial Grant Award. During the first year, we completed the proposed expression profiling of primary DYT1 fibroblasts and controls, while also extending it to a second cell type (lymphoblasts) in collaboration with Dr. Laurie Ozelius (Mt. Sinai). Simultaneous analysis of these gene expression signatures has revealed candidate compounds which may rescue the DYT1 cellular state. Here we propose to take the next step, which will be to confirm the predictions based on the gene expression data via independent biological methods. That information will help us prioritize the best candidates to move forward into animal models and further pre-clinical development.

Dystonia Coalition Obtains ORD Grant
Unprecedented monies to support dystonia research

The Dystonia Coalition is a collaboration of scientists, institutions, and patient organizations formed to advance the pace of clinical research for the dystonias.

In October, Officials at the Office of Rare Disorders (ORD) at the National Institutes of Health (NIH) announced the funding of a five year award for the Dystonia Coalition to advance clinical research on primary focal dystonias including cervical dystonia, spasmodic dysphonia/laryngeal dystonia, blepharospasm, and others. Leading the Coalition will be H. A. Jinnah, MD, PhD, Professor of Neurology and Human Genetics at Emory University in Atlanta, Georgia.

The $6.2 million award will allow the Dystonia Coalition to cultivate a better understanding of the primary focal dystonias and find better therapies. This includes projects to develop a better understanding of their natural history, establish instruments for monitoring symptom severity in clinical trials, and develop proper diagnostic criteria. The creation of a biorepository to store biological samples to support future research is also planned, which will make these resources available to investigators worldwide.

American Dystonia Society

• Bachmann-Strauss Dystonia & Parkinson Foundation
• Beat Dystonia
• Benign Essential Blepharospasm Research Foundation
• DySTonia, Inc.
• Dystonia Medical Research Foundation
• National Spasmodic Dysphonia Association
• National Spasmodic Torticollis Association
• Tyler's Hope for a Dystonia Cure
• We Move

Awarded the Dr. Edward V. Staab Memorial Grant for 2009-2010

1. Accelerating drug discovery for dystonia using chemical genomics

David Christopher Bragg, Ph.D.

Instructor in Neurology

Harvard Medical School

Massachusetts General Hospital  

2.   Developing a Cell-Free Assay of TorsinA Activity

 Rose Goodchild, Ph.D.

 Department of Biochemistry & Cellular and Molecular Biology

University of Tennessee Knoxville

 3. Treatment of Dystonias with Deep Brain Stimulation of Cerebellar Structures

 Robert S. Raike, PhD

Post Doctoral Fellow

Department of Pharmacology

Emory University School of Medicine

Biofocus Drug Discovery

 "Project Firesky" It is estimated that this work, in three phases, will cost between $1,500,000 to $1,750,000. Phase I is complete and Phase II is in the works. Tyler's Hope has joined forces with the DMRF to fund these drug discoveries. This is a very exciting project with a good potential to discover a drug that will treat or eliminate dystonia symptoms. Thank you to the DMRF for paving the way. We are very happy to assist financially to dicover these drugs and fund this research.

Create an rAAV-based animal model for DYT1 dystonia

Ron Mandel, University of FloridaCollege of Medicine

Genetically modified mice are the current gold-standard to model genetic neurological disorders. Indeed, in many cases, these mouse models have been invaluable and extremely reliable. However, there are also notable failures. The most accessible example comes from inherited forms of Parkinson disease (PD). The first PD causing gene was α-synuclein (α-syn) in which point mutations at 2 different amino-acids as well as a triplication of the gene all cause familial PD. Although approximately 20 transgenic and knock-in mice harboring α-syn mutations have been created, none have pathology resembling human PD. A similar pattern is true of the PD causing genes, parkin, DJ-1, and PINK1 for which mouse models do not show PD-like pathology.

In contrast, the use of a viral vector, recombinant adeno-associated virus (rAAV) has provided the only faithful α-syn animal model available today. Certain versions of rAAV vectors (rAAV2 and rAAV5) have the fortuitous property, that, when injected in the substantia nigra of a rodent or a primate, have a relative specificity for infecting dopamine neurons. Thus, when rAAV vectors expressing the human mutated form of α-syn in the substantia nigra of rats, we observed a progressive loss of nigral neurons over approximately 8 weeks. Moreover, as an interesting control we injected the wild-type (un-mutated) of human α-syn and saw similar neuropathology. It was only later that a family with inherited PD was found to have a triplication in the α-syn gene and therefore expressed more of the protein. Another important advantage of the rAAV system is that it can also be injected in any mammalian species. To determine the fidelity of our PD model, we also injected the same α-syn vectors in the marmoset, a small new-world monkey. The nigral α-syn-induced pathology was nearly identical to that seen in the rat model.  We have followed these studies with further studies that have identified proteins that interact with α-syn in the substantia nigra and have also identified alterations to the α-syn that render it non-toxic to dopamine neurons.

Based on these previously successful models based on rAAV, and the lack of useful phenotype in DYT1 mutant or knock-out mice. We propose to make 3 rAAV viruses: wild-type DYT1, mGAGDYT1, and siRNAs against rat DYT1.  Pedro Gonzales-Allegre has graciously agreed to provide us with the constructs for all these proposed viruses. Because we do not know the precise anatomical location of the neuropathology in DYT1, we would propose to inject each of the viruses in different groups of rats including another control virus (green fluorescent protein, GFP) bilaterally in the striatum and globus pallidus, the cerebellum, and both areas together. This would result in 9 groups of rats (n = 8 each). Expression of the proteins takes about 2 weeks to reach a peak so we would begin behavioral testing approximately 3 weeks after vector injection and monthly thereafter. The transgenes are expressed for the life of the animal.

We would first test spontaneous behavior in a battery of tests such as gait analysis, rotorod, and the cylinder rearing test. If we see no behavioral impairments in these tests it will be possible to test the animals in drug paradigms that are known to induce dystonias in rats except that we would use regimens that are mild enough that they would not produce dystonias in normal rats. Possibilities include physostigmine treatment or repeated L-dopa treatment.

This experimental design in rats has the potential to identify the relative contribution of different anatomical structures to dystonias (basal ganglia vs. cerebellum), to help determine if there is a dominant negative effect vs. a loss of function (gagDYT1 vs. DYT1 siRNA). The performance of this experiment including analysis of the data is estimated to be 1 year.

The results of these initial studies would guide future development of the rat model. However, regardless of whether a phenotype is detected or not. It seems prudent to attempt a parallel experiment in marmosets which are available here at UF.  As pointed out in your recent “think tank”, DYT1 dystonia might be a disorder only seen in primates which, among many other differences in physiology, definitely have different anatomical structures that subserve motor function. However, with regard to marmosets, we will come back to the Tyler’s Hope foundation with a detailed plan for research in marmosets.

It is worth pointing out, that if rAAV-mediated over-expression or knock-down of DYT1 causes dystonia in either rats or marmosets, it is then possible to perform experiments whereby the vector is injected in more discrete anatomical areas to potentially identify if different phenotypic forms of dystonia are mediated by different anatomical regions. This last goal would most likely best be reached by studying primates.

It was universally agreed at the “think tank” that prior to beginning drug trials or finding therapeutic avenues, a phenotypically relevant animal model was crucial. Because there has been little or no success in genetic mouse models, we believe that the viral vector approach may provide an animal model for DYT1 dystonia.

“Unraveling torsinA function and dysfunction”

Investigator: William Dauer, MD, Columbia University, New York

DYT1 dystonia is an autosomal dominant disease caused by the loss of a single amino acid in the protein torsinA. We have found that the disease mutation appears to block the normal function of torsinA. Because of this, we believe that experiments to study what happens to neurons when they lose torsinA function will provide clues regarding what goes wrong in the disease. To pursue this research goal, we propose a range of studies on the brains and cultured nerve cells from mice that either lack the torsinA protein or only make the disease form of torsinA. We also believe that learning more about the function of proteins that torsinA interacts with will yield clues to the function of torsinA, and have also proposed experiments along these lines.

Cure Dystonia Initiative Grant
“Therapeutic RNA interference for DYT1 dystonia”
Investigator: Pedro Gonzalez-Alegre, MD, University of Iowa

Pedro Gonzalez-Alegre, MD and his colleagues are using RNAi to restore human neural cells to normal by selectively silencing a gene that causes the movement disorder DYT1 dystonia. The findings also may help researchers apply RNAi to other brain diseases such as Huntington's disease.

Book The Dystonia Patient  funded by Tyler's Hope

Written by the dystonia team at the University of Florida Movement Disorders Center, this book is designed as a practical and complete guide to the integrated management of the dystonia patient. It provides a current understanding of this common but often poorly appreciated condition and a framework for delivering comprehensive multi and intra-disciplinary care. Individual chapters review medical and surgical strategies, botulinum toxin therapy, and programming issues for deep brain stimulators. The remainder of the book is devoted to the important role of health professionals from various disciplines and what the physician needs to know to direct a successful long-term care team for dystonia patients. Features include:      

• Emphasis on a multi-disciplinary team approach to long-term management
• Coverage of both adult and pediatric patients
• Pearls and practical points within each chapter to guide the practitioner
• Lists of resources and referral information for patients, families, and medical teams

   

   

  Okun's remarkable 248 page practical guide, The Dystonia Patient, is the first book that exclusively focuses on patients diagnosed with the rare movement disorder, dystonia. It is well written, efficiently organized with plenty of gems and advice for healthcare providers, who provide direct care to this complex patient population. Most importantly, it highlights the role and application of the multidisciplinary care model for these patients; an under-emphasized care model that is intensely needed for these patients and their families. Many require assistance from a variety of health providers including the physical therapist, occupational therapist, psychologist, pain management specialists, neurosurgeons, nurse practitioners etc.

  Okun has truly given much thought to the total concept of "holistic ", "gestalt" care for these patients when writing this guide. It is quite obvious that the information, provided in a positive,easy-to-read approach, has been thoroughly researched in collaboration with other healthcare providers. I highly applaud the author, a practicing physician specializing in movement disorders, for recognizing the needs and the care required by these patients.

  I highly recommend this book for nursing and medical students, newly practicing neurologists including those ready to enter the arena of movement disorders. The book is an absolute necessity in the office- libraries of all those providing care to the unique Dystonia Patient, as well as organizations providing support to patients and their families.  A true Gem of a Read for all !

  Beka Serdans, RN, MS, NP

  Founder

  Care4Dystonia, Inc.

   

  Tyler's Hope Center for Comprehansive Dystonia Care

  A true Inter-Disciplinary Center for Dystonia, where all the services and resources are consolidated in one location; where bench research is immediately brought to the bedside.

   

  The Summit

   

  Gainesville, Florida      McKnight Brain Institute EMail: tylershope@intermed1.com Phone: 352-273-5550 Tyler's Hope & the McKnight Brain Institute presents: TheAnnual Think Tank on Novel Approaches to a Cure for DYT-1 Dystonia. The Dr. Edward V. Staab Memorial Fund will award the first annual grant to the most promising project towards a Dystonia cure.

   

   

   

     

Nicole Calakos, M.D., Ph.D., Duke University Medical Center

Title: Novel high-throughput approach to screening for modifiers of TorsinA pathology

Hypothesis: The identification of modifiers of cellular inclusion pathology caused by mutant TorsinA proteins will provide novel targets to consider for the treatment of DYT1 dystonia.

Dr. Calakos’ group is developing methodology to perform high-throughput screens for modifiers of TorsinA dysfunction. Numerous assays have shown that disease-associated mutant forms of TorsinA behave differently from the normal protein. The Calakos group proposes to use the abnormal characteristics of disease-associated TorsinA to develop a high-throughput assay with high-content readouts to screen for modifiers that will reduce the abnormal protein burden.  The aim of this work will be to identify new candidate small molecules and pathways for the correction of pathological TorsinA activity. The ultimate goal of this work is to provide new opportunities for the treatment and cure of dystonia.

Dr. Ramon Rodriguez UFMDC

Ampicillan Pilot Trial

Dr. Ramon Rodriguez UFMDC

Ampicillan Trial

Dystonia is a disorder characterized by abnormal postures resulting from involuntary muscle contractions. It can be caused by a variety of conditions, including genetic factors. A mutation in a gene called DYT-1 gene was identified as an important contributor for early onset generalized dystonia. This gene produces an abnormal protein called torsinA, believed to play an important role in the abnormal muscle contraction causing dystonia. Although there are treatments for the symptoms of dystonia, at this moment there is no cure. Recently, a study found that by treating a mouse with a similar DYT-1 mutation with ampicillin (an available antibiotic), there were improvements in the mouse’s ability to walk. We will conduct a study to look at safety and tolerability of ampicillin.

At the University of Florida’s MDC, we  will conduct a study to look at safety and tolerability of ampicillin to improve dystonia symptoms under the clinical trial, “Ampicillin for DYT-1:a Pilot Trial for Safety and Efficacy.”   During the clinical trial patients will receive either study drug (ampicillin) or a placebo for a period of four (4) weeks. Then, there will be one (1) week ‘washout period’ with no study-related medications, followed by a switch to either the placebo or ampicillin for another period of four (4) weeks. As a double-blinded study, neither patient nor the investigator will know which pill patients are taking at any given time. Patients will come to our center five (5) times for evaluation of how they are tolerating the medication(s) and evaluation of their dystonia symptoms.

Nicole Calakos, M.D., Ph.D., Duke University Medical Center

Title: Novel high-throughput approach to screening for modifiers of TorsinA pathology

Hypothesis: The identification of modifiers of cellular inclusion pathology caused by mutant TorsinA proteins will provide novel targets to consider for the treatment of DYT1 dystonia.

Dr. Calakos’ group is developing methodology to perform high-throughput screens for modifiers of TorsinA dysfunction. Numerous assays have shown that disease-associated mutant forms of TorsinA behave differently from the normal protein. The Calakos group proposes to use the abnormal characteristics of disease-associated TorsinA to develop a high-throughput assay with high-content readouts to screen for modifiers that will reduce the abnormal protein burden.  The aim of this work will be to identify new candidate small molecules and pathways for the correction of pathological TorsinA activity. The ultimate goal of this work is to provide new opportunities for the treatment and cure of dystonia.

Dystonia International Patient Registry (DIPR)    The Dystonia International Patient Registry (DIPR) is an observational registry of individuals who carry the DYT-1 gene.  The ultimate goal of this registry is to provide  the natural history of DYT-1 Dystonia worldwide.  All scientific and medical investigators are encouraged to utilize the registry as a source of patients for clinical research and as a source of demographic information on the patient population. Click here to access the registry.

 2011 Projects funded:

Patient-specific induced pluripotent stem (iPS) cells as a model for DYT1 dystonia

Nutan Sharma, MD PhD and Cristopher Bragg, PhD

Massachusetts General Hospital/Harvard University

The objective of this research is to establish a collection of DYT1-specific induced pluripotent stem (iPS) cells, which are stem cells derived from patient-donated somatic (skin) cells, and matched controls that will serve as a public resource for the dystonia research community. These cells, which are generated by reprogramming skin fibroblasts resemble embryonic stem cells in culture and can be differentiated into specific cell types, such as neurons, needed for research. Thus, DYT1-specific iPS cells will represent a self-renewing population of cells bearing ‘natural’ levels of the mutated target protein, torsinAΔE, capable of becoming human neurons, which are thought to be critical in developing DYT1 dystonia. This model would enable multiple dystonia research groups to conduct new studies designed to study neuron-specific defects associated with torsinAΔE.

The current plan is to perform an initial analysis of DYT1 vs. control-derived neurons and publish the findings, at which point the cells will be deposited with the Coriell Institute for public distribution. The Institute has already been notified about the intent to deposit the DYT1 iPS cells.

Project FireSky

BioFocus

BioFocus target discovery platform uses human cells to model diseases in combination with functional genomics. BioFocus introduces fragments of genetic material, DNA or RNA into cells using collections of specially designed viruses to study the roles of individual genes in disease processes. Advanced readouts are then used to identify the genes that have potential effect on the disease-related process in this model system. After further validation assays, these targets form the basis for the development of new drugs. This unique approach combines cell assays with high-throughput technology, supported by state-of-the-art monitoring equipment.

Outline of the Project so far

·        The major objective of the Project is to identify genes and proteins that modify the DYT1 dystonia phenotype.

·        To accomplish that a genetic approach is used. It is based on silencing (“turning off”) selected genes by using small RNA molecule specific for individual genes.

·        The RNA molecules target more than 4,500 genes preselected by BioFocus as the most promising, drugable targets.

·        During silencing torsinA function is monitored in cultured cell using a quantitative detection assay developed in Phase 1 of the Project.

·        Several rounds of robotic screening and selection of specifically interfering genes and proteins have been performed in Phase 2.

·        Identified genes and proteins will be potential drug targets for DYT1 and possibly other dystonias.

·        Selected targets will be utilized in subsequent drug discovery programs to develop therapeutics to modify the phenotype of DYT1 dystonia.

Phase 1 – Assay Development - completed

Phase 2 – Screening - ongoing

An assay using the neuron-like cell line to measure the secretion of proteins upon viral ‘suppression’ of torsinA was developed during Phase 1, and it is this assay that is being used during Phase 2 to screen the BioFocus’ SilenceSelect™ library. The SilenceSelect™ library targets the human drugable genome (>4,500 genes that are targeted by 12,000 viruses), which ensures that the identified targets are tractable for small molecule drug discovery.  The screening is done from an independent re-propagation of the viral stocks to confirm activity. Re-screening will reduce the number of hits eliminating some that are produced due to the experimental procedure or secondary effects. Additional toxicity experiments will also be performed to appropriately adjust the screening conditions.

Phase 3 – Target validation

In the last phase of the project, a narrowed down selection of validated target will be screened again by using a different assay and cell biological methods. The goal is to identify a manageable number of validated target that are likely players in the disease process. When successfully completed this will allow for identification of genes and proteins affecting torsinA function that might become targets for rational drug design. The identified targets will also enrich our knowledge about the role of torsinA in neurons and guide future research in this area.

2010 Summit - Dr. Edward V. Staab Memorial Grants

Nicole Calakos, M.D., Ph.D., Duke University Medical Center

Title: Novel high-throughput approach to screening for modifiers of TorsinA pathology

Hypothesis: The identification of modifiers of cellular inclusion pathology caused by mutant TorsinA proteins will provide novel targets to consider for the treatment of DYT1 dystonia.

Dr. Calakos’ group is developing methodology to perform high-throughput screens for modifiers of TorsinA dysfunction. Numerous assays have shown that disease-associated mutant forms of TorsinA behave differently from the normal protein. The Calakos group proposes to use the abnormal characteristics of disease-associated TorsinA to develop a high-throughput assay with high-content readouts to screen for modifiers that will reduce the abnormal protein burden.  The aim of this work will be to identify new candidate small molecules and pathways for the correction of pathological TorsinA activity. The ultimate goal of this work is to provide new opportunities for the treatment and cure of dystonia.

Dr. Ramon Rodriguez UFMDC

Ampicillan Trial

Dystonia is a disorder characterized by abnormal postures resulting from involuntary muscle contractions. It can be caused by a variety of conditions, including genetic factors. A mutation in a gene called DYT-1 gene was identified as an important contributor for early onset generalized dystonia. This gene produces an abnormal protein called torsinA, believed to play an important role in the abnormal muscle contraction causing dystonia. Although there are treatments for the symptoms of dystonia, at this moment there is no cure. Recently, a study found that by treating a mouse with a similar DYT-1 mutation with ampicillin (an available antibiotic), there were improvements in the mouse’s ability to walk. We will conduct a study to look at safety and tolerability of ampicillin.

At the University of Florida’s MDC, we  will conduct a study to look at safety and tolerability of ampicillin to improve dystonia symptoms under the clinical trial, “Ampicillin for DYT-1:a Pilot Trial for Safety and Efficacy.”   During the clinical trial patients will receive either study drug (ampicillin) or a placebo for a period of four (4) weeks. Then, there will be one (1) week ‘washout period’ with no study-related medications, followed by a switch to either the placebo or ampicillin for another period of four (4) weeks. As a double-blinded study, neither patient nor the investigator will know which pill patients are taking at any given time. Patients will come to our center five (5) times for evaluation of how they are tolerating the medication(s) and evaluation of their dystonia symptoms.

D. Cristopher Bragg, Ph.D.

Massachusetts General Hospital

Harvard Medical School

 “Accelerating drug discovery for dystonia using chemical genomics” which has been supported in the past year by the Dr. Edward V. Staab Memorial Grant Award. During the first year, we completed the proposed expression profiling of primary DYT1 fibroblasts and controls, while also extending it to a second cell type (lymphoblasts) in collaboration with Dr. Laurie Ozelius (Mt. Sinai). Simultaneous analysis of these gene expression signatures has revealed candidate compounds which may rescue the DYT1 cellular state. Here we propose to take the next step, which will be to confirm the predictions based on the gene expression data via independent biological methods. That information will help us prioritize the best candidates to move forward into animal models and further pre-clinical development.

Dystonia Coalition Obtains ORD Grant
Unprecedented monies to support dystonia research

The Dystonia Coalition is a collaboration of scientists, institutions, and patient organizations formed to advance the pace of clinical research for the dystonias.

In October, Officials at the Office of Rare Disorders (ORD) at the National Institutes of Health (NIH) announced the funding of a five year award for the Dystonia Coalition to advance clinical research on primary focal dystonias including cervical dystonia, spasmodic dysphonia/laryngeal dystonia, blepharospasm, and others. Leading the Coalition will be H. A. Jinnah, MD, PhD, Professor of Neurology and Human Genetics at Emory University in Atlanta, Georgia.

The $6.2 million award will allow the Dystonia Coalition to cultivate a better understanding of the primary focal dystonias and find better therapies. This includes projects to develop a better understanding of their natural history, establish instruments for monitoring symptom severity in clinical trials, and develop proper diagnostic criteria. The creation of a biorepository to store biological samples to support future research is also planned, which will make these resources available to investigators worldwide.

American Dystonia Society

• Bachmann-Strauss Dystonia & Parkinson Foundation
• Beat Dystonia
• Benign Essential Blepharospasm Research Foundation
• DySTonia, Inc.
• Dystonia Medical Research Foundation
• National Spasmodic Dysphonia Association
• National Spasmodic Torticollis Association
• Tyler's Hope for a Dystonia Cure
• We Move

Awarded the Dr. Edward V. Staab Memorial Grant for 2009-2010

1. Accelerating drug discovery for dystonia using chemical genomics

David Christopher Bragg, Ph.D.

Instructor in Neurology

Harvard Medical School

Massachusetts General Hospital  

2.   Developing a Cell-Free Assay of TorsinA Activity

 Rose Goodchild, Ph.D.

 Department of Biochemistry & Cellular and Molecular Biology

University of Tennessee Knoxville

 3. Treatment of Dystonias with Deep Brain Stimulation of Cerebellar Structures

 Robert S. Raike, PhD

Post Doctoral Fellow

Department of Pharmacology

Emory University School of Medicine

Biofocus Drug Discovery

 "Project Firesky" It is estimated that this work, in three phases, will cost between $1,500,000 to $1,750,000. Phase I is complete and Phase II is in the works. Tyler's Hope has joined forces with the DMRF to fund these drug discoveries. This is a very exciting project with a good potential to discover a drug that will treat or eliminate dystonia symptoms. Thank you to the DMRF for paving the way. We are very happy to assist financially to dicover these drugs and fund this research.

Create an rAAV-based animal model for DYT1 dystonia

Ron Mandel, University of FloridaCollege of Medicine

Genetically modified mice are the current gold-standard to model genetic neurological disorders. Indeed, in many cases, these mouse models have been invaluable and extremely reliable. However, there are also notable failures. The most accessible example comes from inherited forms of Parkinson disease (PD). The first PD causing gene was α-synuclein (α-syn) in which point mutations at 2 different amino-acids as well as a triplication of the gene all cause familial PD. Although approximately 20 transgenic and knock-in mice harboring α-syn mutations have been created, none have pathology resembling human PD. A similar pattern is true of the PD causing genes, parkin, DJ-1, and PINK1 for which mouse models do not show PD-like pathology.

In contrast, the use of a viral vector, recombinant adeno-associated virus (rAAV) has provided the only faithful α-syn animal model available today. Certain versions of rAAV vectors (rAAV2 and rAAV5) have the fortuitous property, that, when injected in the substantia nigra of a rodent or a primate, have a relative specificity for infecting dopamine neurons. Thus, when rAAV vectors expressing the human mutated form of α-syn in the substantia nigra of rats, we observed a progressive loss of nigral neurons over approximately 8 weeks. Moreover, as an interesting control we injected the wild-type (un-mutated) of human α-syn and saw similar neuropathology. It was only later that a family with inherited PD was found to have a triplication in the α-syn gene and therefore expressed more of the protein. Another important advantage of the rAAV system is that it can also be injected in any mammalian species. To determine the fidelity of our PD model, we also injected the same α-syn vectors in the marmoset, a small new-world monkey. The nigral α-syn-induced pathology was nearly identical to that seen in the rat model.  We have followed these studies with further studies that have identified proteins that interact with α-syn in the substantia nigra and have also identified alterations to the α-syn that render it non-toxic to dopamine neurons.

Based on these previously successful models based on rAAV, and the lack of useful phenotype in DYT1 mutant or knock-out mice. We propose to make 3 rAAV viruses: wild-type DYT1, mGAGDYT1, and siRNAs against rat DYT1.  Pedro Gonzales-Allegre has graciously agreed to provide us with the constructs for all these proposed viruses. Because we do not know the precise anatomical location of the neuropathology in DYT1, we would propose to inject each of the viruses in different groups of rats including another control virus (green fluorescent protein, GFP) bilaterally in the striatum and globus pallidus, the cerebellum, and both areas together. This would result in 9 groups of rats (n = 8 each). Expression of the proteins takes about 2 weeks to reach a peak so we would begin behavioral testing approximately 3 weeks after vector injection and monthly thereafter. The transgenes are expressed for the life of the animal.

We would first test spontaneous behavior in a battery of tests such as gait analysis, rotorod, and the cylinder rearing test. If we see no behavioral impairments in these tests it will be possible to test the animals in drug paradigms that are known to induce dystonias in rats except that we would use regimens that are mild enough that they would not produce dystonias in normal rats. Possibilities include physostigmine treatment or repeated L-dopa treatment.

This experimental design in rats has the potential to identify the relative contribution of different anatomical structures to dystonias (basal ganglia vs. cerebellum), to help determine if there is a dominant negative effect vs. a loss of function (gagDYT1 vs. DYT1 siRNA). The performance of this experiment including analysis of the data is estimated to be 1 year.

The results of these initial studies would guide future development of the rat model. However, regardless of whether a phenotype is detected or not. It seems prudent to attempt a parallel experiment in marmosets which are available here at UF.  As pointed out in your recent “think tank”, DYT1 dystonia might be a disorder only seen in primates which, among many other differences in physiology, definitely have different anatomical structures that subserve motor function. However, with regard to marmosets, we will come back to the Tyler’s Hope foundation with a detailed plan for research in marmosets.

It is worth pointing out, that if rAAV-mediated over-expression or knock-down of DYT1 causes dystonia in either rats or marmosets, it is then possible to perform experiments whereby the vector is injected in more discrete anatomical areas to potentially identify if different phenotypic forms of dystonia are mediated by different anatomical regions. This last goal would most likely best be reached by studying primates.

It was universally agreed at the “think tank” that prior to beginning drug trials or finding therapeutic avenues, a phenotypically relevant animal model was crucial. Because there has been little or no success in genetic mouse models, we believe that the viral vector approach may provide an animal model for DYT1 dystonia.

“Unraveling torsinA function and dysfunction”

Investigator: William Dauer, MD, Columbia University, New York

DYT1 dystonia is an autosomal dominant disease caused by the loss of a single amino acid in the protein torsinA. We have found that the disease mutation appears to block the normal function of torsinA. Because of this, we believe that experiments to study what happens to neurons when they lose torsinA function will provide clues regarding what goes wrong in the disease. To pursue this research goal, we propose a range of studies on the brains and cultured nerve cells from mice that either lack the torsinA protein or only make the disease form of torsinA. We also believe that learning more about the function of proteins that torsinA interacts with will yield clues to the function of torsinA, and have also proposed experiments along these lines.

Cure Dystonia Initiative Grant
“Therapeutic RNA interference for DYT1 dystonia”
Investigator: Pedro Gonzalez-Alegre, MD, University of Iowa

Pedro Gonzalez-Alegre, MD and his colleagues are using RNAi to restore human neural cells to normal by selectively silencing a gene that causes the movement disorder DYT1 dystonia. The findings also may help researchers apply RNAi to other brain diseases such as Huntington's disease.

Book The Dystonia Patient  funded by Tyler's Hope

Written by the dystonia team at the University of Florida Movement Disorders Center, this book is designed as a practical and complete guide to the integrated management of the dystonia patient. It provides a current understanding of this common but often poorly appreciated condition and a framework for delivering comprehensive multi and intra-disciplinary care. Individual chapters review medical and surgical strategies, botulinum toxin therapy, and programming issues for deep brain stimulators. The remainder of the book is devoted to the important role of health professionals from various disciplines and what the physician needs to know to direct a successful long-term care team for dystonia patients. Features include:      

• Emphasis on a multi-disciplinary team approach to long-term management
• Coverage of both adult and pediatric patients
• Pearls and practical points within each chapter to guide the practitioner
• Lists of resources and referral information for patients, families, and medical teams

   

   

  Okun's remarkable 248 page practical guide, The Dystonia Patient, is the first book that exclusively focuses on patients diagnosed with the rare movement disorder, dystonia. It is well written, efficiently organized with plenty of gems and advice for healthcare providers, who provide direct care to this complex patient population. Most importantly, it highlights the role and application of the multidisciplinary care model for these patients; an under-emphasized care model that is intensely needed for these patients and their families. Many require assistance from a variety of health providers including the physical therapist, occupational therapist, psychologist, pain management specialists, neurosurgeons, nurse practitioners etc.

  Okun has truly given much thought to the total concept of "holistic ", "gestalt" care for these patients when writing this guide. It is quite obvious that the information, provided in a positive,easy-to-read approach, has been thoroughly researched in collaboration with other healthcare providers. I highly applaud the author, a practicing physician specializing in movement disorders, for recognizing the needs and the care required by these patients.

  I highly recommend this book for nursing and medical students, newly practicing neurologists including those ready to enter the arena of movement disorders. The book is an absolute necessity in the office- libraries of all those providing care to the unique Dystonia Patient, as well as organizations providing support to patients and their families.  A true Gem of a Read for all !

  Beka Serdans, RN, MS, NP

  Founder

  Care4Dystonia, Inc.

   

  Tyler's Hope Center for Comprehansive Dystonia Care

  A true Inter-Disciplinary Center for Dystonia, where all the services and resources are consolidated in one location; where bench research is immediately brought to the bedside.

   

  The Summit

   

  Gainesville, Florida      McKnight Brain Institute EMail: tylershope@intermed1.com Phone: 352-273-5550 Tyler's Hope & the McKnight Brain Institute presents: TheAnnual Think Tank on Novel Approaches to a Cure for DYT-1 Dystonia. The Dr. Edward V. Staab Memorial Fund will award the first annual grant to the most promising project towards a Dystonia cure.

   

   

   

     

Nicole Calakos, M.D., Ph.D., Duke University Medical Center

Title: Novel high-throughput approach to screening for modifiers of TorsinA pathology

Hypothesis: The identification of modifiers of cellular inclusion pathology caused by mutant TorsinA proteins will provide novel targets to consider for the treatment of DYT1 dystonia.

Dr. Calakos’ group is developing methodology to perform high-throughput screens for modifiers of TorsinA dysfunction. Numerous assays have shown that disease-associated mutant forms of TorsinA behave differently from the normal protein. The Calakos group proposes to use the abnormal characteristics of disease-associated TorsinA to develop a high-throughput assay with high-content readouts to screen for modifiers that will reduce the abnormal protein burden.  The aim of this work will be to identify new candidate small molecules and pathways for the correction of pathological TorsinA activity. The ultimate goal of this work is to provide new opportunities for the treatment and cure of dystonia.

Dr. Ramon Rodriguez UFMDC

Ampicillan Pilot Trial

Dr. Ramon Rodriguez UFMDC

Ampicillan Trial

Dystonia is a disorder characterized by abnormal postures resulting from involuntary muscle contractions. It can be caused by a variety of conditions, including genetic factors. A mutation in a gene called DYT-1 gene was identified as an important contributor for early onset generalized dystonia. This gene produces an abnormal protein called torsinA, believed to play an important role in the abnormal muscle contraction causing dystonia. Although there are treatments for the symptoms of dystonia, at this moment there is no cure. Recently, a study found that by treating a mouse with a similar DYT-1 mutation with ampicillin (an available antibiotic), there were improvements in the mouse’s ability to walk. We will conduct a study to look at safety and tolerability of ampicillin.

At the University of Florida’s MDC, we  will conduct a study to look at safety and tolerability of ampicillin to improve dystonia symptoms under the clinical trial, “Ampicillin for DYT-1:a Pilot Trial for Safety and Efficacy.”   During the clinical trial patients will receive either study drug (ampicillin) or a placebo for a period of four (4) weeks. Then, there will be one (1) week ‘washout period’ with no study-related medications, followed by a switch to either the placebo or ampicillin for another period of four (4) weeks. As a double-blinded study, neither patient nor the investigator will know which pill patients are taking at any given time. Patients will come to our center five (5) times for evaluation of how they are tolerating the medication(s) and evaluation of their dystonia symptoms.

Nicole Calakos, M.D., Ph.D., Duke University Medical Center

Title: Novel high-throughput approach to screening for modifiers of TorsinA pathology

Hypothesis: The identification of modifiers of cellular inclusion pathology caused by mutant TorsinA proteins will provide novel targets to consider for the treatment of DYT1 dystonia.

Dr. Calakos’ group is developing methodology to perform high-throughput screens for modifiers of TorsinA dysfunction. Numerous assays have shown that disease-associated mutant forms of TorsinA behave differently from the normal protein. The Calakos group proposes to use the abnormal characteristics of disease-associated TorsinA to develop a high-throughput assay with high-content readouts to screen for modifiers that will reduce the abnormal protein burden.  The aim of this work will be to identify new candidate small molecules and pathways for the correction of pathological TorsinA activity. The ultimate goal of this work is to provide new opportunities for the treatment and cure of dystonia.