Grant Award Recipients
KDA RESEARCH GRANTS
As of November, 2017 the KDA has awarded $1,033,214 in research grants to help find a cure or treatment for Kennedy's Disease.
2017 - Five research grants were awarded totaling $199,000. Award recipients and summaries are listed below.
Targeting AR toxicity in SBMA by modulation of USP7 activity.
Anna Pluciennik, Ph.D.
Department of Biochemistry and Molecular Biology
Thomas Jefferson University
Summary: PolyQ-expanded androgen receptor-dependent cellular toxicity in the neuromuscular system is a characteristic feature of SBMA pathogenesis, although the molecular mechanisms for these effects are poorly understood. However, because cellular toxicity in SBMA is likely to arise, at least in part, from the polyQ-expanded-dependent dysregulation of protein-protein interactions that sustain normal cellular function, we reasoned that identification of such dysregulated interactions might help reveal potential therapeutic targets for disease modification. Therefore, as part of a KDA-funded project (2014), we used a quantitative proteomics approach and identified a deubiquitinating enzyme, USP7, that preferentially interacts with polyQ-expanded AR and contributes to toxicity. In fact, we have demonstrated that not only does partial knockdown of USP7 protein expression decrease mutant AR-dependent aggregation and dihydrotestosterone-dependent cytotoxicity, but also that overexpression of the protein aggravates these cellular effects. These results establish the need for further investigation into the role of the deubiquitinating function of USP7 in SBMA pathogenesis. The objective of this -research is to investigate the functional consequence of pharmacological inhibition of USP7 deubiquitinase activity in cellular and mouse models of SBMA. These studies will extend our previous KDA-funded proteomics work and, if successful, will establish the validity of inhibition of USP7 activity as a therapeutic approach for the treatment of SBMA.
Identification of New Polyglutamine-Specific Mutant AR-Interacting Proteins in SBMA Motor Neurons
Xia Feng, PhD.
Postdoctoral Fellow, Neurogenetics Branch
National Institute of Neurological Disorders and Stroke, National Institute of Health
Summary: Spinal and bulbar muscular atrophy (SBMA) is a slowly progressive neuromuscular disease. As the disease proceeds, nerve cells in the spinal cord (called motor neurons) start to die and muscle cells will waste away. The causative mutation of SBMA is the mutant androgen receptor (AR) with an abnormal expansion in the certain region. Such aberrant expansion in the mutant protein (called a polyglutamine tract) damages the normal function of the protein as well as obtains toxicity. Thus, it is important to understand how the mutant AR is regulated via other proteins in the disease. Here, I propose to use a human induced pluripotent stem cell model to generate disease-relevant motor neuron-like cells, and use these cells to identify specific interacting proteins of the mutant androgen receptor. I am hoping that my research will help provide a motor neuron-specific basis for designing and developing novel therapeutics for the treatment of SBMA.
Transcellular regulation of the proteostasis network in Kennedy’s disease
Laura Bott, Ph.D.
Molecular Biosciences, Northwestern University
Summary: Misfolding and abnormal accumulation of the mutant androgen receptor in Kennedy’s disease indicates that the cellular machinery controlling protein abundance, folding, and transport (proteostasis) may be defective in the disease. We are planning to study molecular mechanisms of Kennedy’s disease in the worm Caenorhabditis elegans. For this, we will genetically engineer worms to express the mutant androgen receptor in the neuromuscular system and assess effects of the polyglutamine expansion on proteostasis regulation in this model organism. Insights into cell type-specific effects and regulation across tissues may lead to new therapeutic approaches for this disease.
One gene, many proteins: investigating the role of AR isoform 2 as a therapeutic target for SBMA
Carlo Rinaldi, MD, PhD
Department of Physiology, Anatomy and Genetics
University of Oxford
Summary: The activity of steroid hormone receptors, such as progesterone and oestrogen receptors, is modulated by a number of isoforms and splice variants in a tissue-specific manner, in both health and disease. Androgen receptor may be no exception. AR isoform 2, or AR45 by the molecular weight of its encoded widely-expressed protein, is the only naturally occurring AR isoform, arising from use of an alternative transcriptional start site in intron 1 of the AR gene and containing a short, unique seven amino-acid-long N-terminal stretch instead of the long N-terminal domain found in the full length AR. Overarching aim of this proposal is to unravel the contribution of AR isoform 2 to SBMA pathogenesis and provide a novel therapeutic target for this disease, suitable for oligonucleotide antisense targeting without running the risks associated with silencing of the only available copy of the AR gene in males.
The use of induced stem cells and microfluidics for developing new assays to identify new therapies for Kennedy’s disease
Thomas M Durcan, Ph.D., Assistant Professor,
Montreal Neurological Institute, McGill University The use of induced stem cells and microfluidics for developing new assays to identify new therapies for Kennedy’s disease
Summary: It is important to explore new avenues in the search for treatments for Kennedy’s disease (KD). Our overall goal is to develop disease-relevant assays that use human motor neurons made from stem cells of both men with Kennedy’s disease and healthy individuals. Using special silicone microfluidic devices to grow motor neurons, we will measure the function of neurons upon exposure to different compounds. First, we will develop assays to measure the survival and growth of the motor neurons. Next, we will look at the how well mitochondria work and move within the neurons. Finally, we aim to set up an assay to test the ability of the normal and KD motor neurons to form synapses (junctions). Taken together, these innovative assays will provide a foundation to build a KD drug discovery platform to screen for promising compounds and targets to treat KD.
2016 - Three research grants were awarded totaling $150,000. Each grant recipient recieved $50,000 each.
Dr Bilal Malik
Professor Linda Greensmith’s Lab, UCL, Institute of Neurology, UK
Targeting pathways of disease in Spinal Bulbar and Muscular Atrophy (SBMA)
Spinal and Bulbar Muscular Atrophy (SBMA), also known as Kennedy’s disease (KD), is adult-onset slowly progressing rare inherited neuromuscular disorder that primarily affects males. As yet there are no effective treatments that can cure the disease or delay its progression. The disease is primarily characterised by muscle weakness and wasting, and degeneration of motor neuron cells within the spinal cord and brain.
Our aim is to establish why motor neurons and muscles degenerate in SBMA by investigating the genes and pathways that underlie disease. The identification of changes that occur early in disease may identify the mechanisms responsible for disease and help establish novel therapeutic targets. This proposal offers the unique opportunity to undertake a comparative study of two platforms that model SBMA, each with its own merits: i) a well-characterised mouse model in which muscle and motor neurons can be examined at various stages of disease, and ii) human cell models, including stem-cell derived motor neurons and patient muscle cells acquired from biopsies. By comparing and contrasting the changes in gene expression in these models of the specific cells affected in SBMA we hope to identify the key changes in gene expression that take place early in disease, identifying a common signature in the pathways of pathology. The results of this study will not only help define novel therapeutic targets with a greater level of confidence by analysing several complimentary models of SBMA, but also allow us to test treatment strategies in a human cell model of the disease.
Dr. Janghoo Lim
Yale University School of Medicine
The role of VCP in the pathogenesis of Kennedy's disease
Spinal and Bulbar Muscular Atrophy (SBMA; Kennedy’s Disease) is a neuromuscular disease
that affects motor neurons and skeletal muscles. The symptoms of SBMA include progressive
weakness of the limbs and facial muscles, as well as difficulty with speaking and swallowing.
SBMA is an X-linked disease that primarily affects men, and is caused by a polyglutamine
expansion in the gene Androgen Receptor (AR). The polyglutamine expansion in AR makes the
protein toxic, and can lead to the formation of protein aggregates inside of cells as well as cell
death. SBMA is one of nine different polyglutamine expansion disorders that are linked to
neurodegeneration. How polyglutamine expanded AR causes SBMA is still being studied, and
there are no effective therapeutics available. In order to better understand the mechanisms that
cause SBMA and translate these results into the development of effective therapeutics, my
proposal aims to assess how the protein Valosin-Containing Protein (VCP) is involved in SBMA.
VCP plays a role in breaking down mutant or damaged proteins, and has been studied in other
neurodegenerative disorders. Based on our preliminary data, we hypothesize that VCP can
regulate the expression and/or the activity of polyglutamine expanded AR. Our proposal will
examine how VCP affects the development of protein aggregates and cell death in SBMA. We
will use cell culture models and fruit flies, both of which are commonly used to study SBMA.
This research will help develop a more thorough understanding of what causes SBMA, and
provide important information when developing new therapeutics for this devastating disease.
Manuela Basso, Ph.D.
Laboratory of Transcriptional Neurobiology
Centre for Integrative Biology
University of Trento
Insights into the molecular pathology of SBMA: Targeting PRMT6 to attenuate the disease.
In collaboration with the Laboratory of Dr. Pennuto, we have recently discovered that a protein, called PRMT6, exacerbates the toxicity induced by mutant androgen receptor, while its inhibition rescues it in cells and flies. Our strategy is to develop a therapy that preserves AR physiological functions while abolishing the toxicity acquired upon polyglutamine expansion. Thus, we propose to silence PRMT6 both via selective pharmacological inhibitors and via gene-silencing to choose the best system to move our studies in pre-clinical models.
provide important information when developing new therapeutics for this devastating disease.
2015 - Two research grants were awarded totaling $100,000
Project Title - 'Insights into the Molecular Pathology of SBMA'
Miltiadis Paliouras, Ph.D. – Principal Investigator
Assistant Professor, Department of Medicine, McGill University
We have recently undertaken a proteomics study, coupled to a comparative systems biology analysis to characterize the protein composition of polyQ-Androgen receptor (polyQ-AR) complexes vs. wild-type-AR (WT-AR) complexes. The data shows that AR-interacting partners change dynamically between polyQ-AR and the WT-AR. Our initial results indicate that the polyQ-AR associates with several RNA-binding proteins that participate in RNA metabolism, presumably through an alternative RNA-splicing pathway. The possibility that a transcription factor protein like the AR could also bind to RNA is very plausible; and has been previously described by pioneers in the AR field.
We have employed a unique and powerful genetic approach of using a “humanized” polyQ-AR Drosophila (fly) model that allows us to ectopically express the polyQ-AR in the developing fly embryo. By crossing polyQ-AR flies to RNAi flies of homologous genes encoding candidate polyQ-AR RNA-binding protein interactors, we have been able to identify a number of ligand-dependent enhancers and suppressors.
As neuronal survival has been area of investigation for trinucleotide repeat diseases, and is thought to contribute to disease etiology and pathology, we have specifically focused on mitochondrial dysfunction and apoptosis, and proteasome inhibition. We will study the impact of overexpressing and silencing selected interactors from our fly genetic interaction screen, on cellular processes associated with SBMA such as androgen ligand- and polyQ-AR-induced mitochondrial dysfunction and proteasome inhibition. To do so, we will be using mouse motor neuron-derived cell lines expressing 65Q- and 24Q-AR, and transfected HEK-GFPu cells, respectively.
Our approach of combining proteomics and systems biology is extremely well suited to unraveling complex biological systems and has already proven extremely useful. In the future, we will be interested how polyQ-AR would also impact on the global alternative RNA splicing profile of SBMA patients, and determine the correlation to neuronal survival. These experiments will enable us to infer potentially new molecular mechanisms of polyQ-AR function and give us insight into the mechanisms underlying neurodegeneration in SBMA.
Project Title: ‘Characterizing the AR-TFEB Interactome in SBMA’.
Constanza J. Cortes, PhD
University of California, San Diego (UCSD)
Sanford Consortium for Regenerative Medicine
Protein-protein interactions are one of the key regulatory steps to normal protein function. Understanding how these protein complexes are altered with mutant AR versus normal AR is key to uncover new mechanisms of mutant AR toxicity in Kennedy’s Disease. In particular, I have previously shown that in Kennedy’s disease, AR can switch between activating or inhibiting a cellular recycling process known as autophagy by altering its interaction with novel co-regulator Transcription Factor E-B (TFEB). In this project, I will define the interacting partners of the AR-TFEB duplex, with the ultimate goal of identifying novel proteins that regulate this molecular switch and have important therapeutic potential for KD.
2014 - Two research grants were awarded totaling $100,000
Anna Pluciennik, Ph.D., a researcher at Dr. Diane Merry's Lab, Department of Biochemistry and Molecular Biology, Thomas Jefferson University
Amount Awarded: $50,000
Helen Cristina Miranda, postdoctoral researcher at Al La Spada's Lab, University of California, San Diego
Amount Awarded: $50,000
2013 - Two research grants were awarded totaling $100,000
Amount Awarded: $50,000
2012 - One research grant was awarded for $25,000
Jamie A. Johansen, Ph.D., Central Michigan University, Mount Pleasant, MI
Amount Awarded: $25,000Proposal: "Mechanisms of anti-androgen treatment in SBMA mice muscles"
-2011 - Three research grants were awarded totaling $65,000
Thanks to the generosity of its supporters, the Board of Directors of the Kennedy’s Disease Association announced today that they awarded three research grants. The three recipients and a brief explanation of their research are shown below.
1. Masahisa Katsuno, M.D. – Ph.D., Department of Neurology, Nagoya University Graduate School of Medicine
Amount Awarded: $25,000
Proposal: Elucidation of neuronal death signaling pathways and development of disease-modifying therapies for Kennedy’s disease
Brief Explanation: Their lab has evidence that the synthesis of two proteins are affected by the defective androgen receptor in KD. They wish to determine if neuronal cell death is caused by the alteration in the levels of these proteins and if cell death canbe prevented by the addition of drugs that target the activity of these proteins.
2. Elise Kikis, Ph. D., Northwestern University
Amount Awarded: $20,000
Proposal: Modeling SBMA: from understanding proteotoxicity to identifying therapeutics
Brief Explanation: They believe that the specific cell death is due to the accumulation of misfolded proteins (the androgen receptor) and the inability of cells to handle this accumulation. They propose to use a new model organism (a little worm called C. elegans – a very common and important model system in biology) to examine how different cell types handle the misfolded proteins and genetically look for other proteins that may help the cell get rid of the messed up proteins.
3. Sara Parodi, Ph.D., Department of Neuroscience and Brain Technologies, Genoa, Italy
Amount Awarded: $20,000
Proposal: Identification of PKA signaling as a new therapeutic approach for SBMA
Brief Explanation: There is evidence that the cell death may involve changes to the androgen receptor (specifically changes in which phosphate is added to the protein, a process called phosphorylation). They hope to determine whether this cell death can be stopped due to the activation of another protein, known as PKA.-
2009 - 3 research grants were awarded totaling $50,000
#1. $25,000 Parsa Kazemi-Esfarjani, B.Sc., Ph.D. is a researcher for the Department of Pediatrics, Division of Genetics, Institute for Genomic Medicine at the School of Medicine, University of California, San Diego
Essentially this research will attempt to determine whether or not, and to what extent, the mutant androgen receptor (AR) in the muscle contributes to both muscle atrophy and motor neuron degeneration in Kennedy’s Disease (KD). They have produced a mouse model of KD in which they can remove the mutant AR gene just in the muscle cells (this is due to a genetic manipulation), leaving the mutant AR intact in the rest of the cells. This type of research will hopefully show us which cells and tissues are the most important for the timing of the appearance of KD symptoms (i.e., the onset) and/or the pace of their progression. With this knowledge, we will be able to develop our therapies for KD more effectively and target them to the appropriate tissues.
#2 $20,000 Maria Pennuto,Ph.D. Department of Neuroscience, Italian Institute of Technology, Genova, Italy.
This research is based on results that Maria found when she worked in Dr. Fischbeck's lab at NIH. She had found that a specific modification of the mutant androgen receptors (AR) results in decreased toxicity and that this modification was due to the activation of an enzyme known as PKA. She intends to investigate more thoroughly the relationship between PKA and toxicity and to search for drugs that may activate PKA and thus may lessen the effects of the mutant AR.
#3 $10,000 Lenore Beitel, Ph.D. Lady Davis Institute for Medical Research, SMBD-Jewish General Hospital, Montreal, Quebec, Canada
This research is to further study the role of the proteasome and its ability (or lack of ability) to degrade mutant androgen receptors (AR). She hopes to determine directly if the mutant AR really does 'clog' up the proteasome. Up to this point, most of the evidence for such an effect is circumstantial.
2008 - One $ 50,000 research grants was awarded:
At the Kennedy's Disease Association's Annual Conference and Research Symposium in November, Board Member Ed Meyertholen awarded Heather Montie, Ph.D., with a $50,000 check to support her Kennedy's Disease research at Thomas Jefferson University in Philadelphia. This grant will fund research that will investigate the androgen receptor (AR) - this is the protein that is altered in Kennedy's Disease (KD). The AR mediates all the effects of testosterone in cells. Without the AR, testosterone will have no effect. Normally, the AR is activated by testosterone. Once activated by testosterone, the AR causes changes in the activity of the cell. It is believed that in cells from those with KD, the cell is not capable of removing the 'old' AR that has been used and this somehow results in the death of the cells. Preliminary research by Dr. Montie and her colleagues have indicated that the KD form of AR is chemically altered by the cell through a process of hyperacetylation and it is possible that if this process is blocked, the cell may be more capable of removing the AR and thus should not die. The KDA grant will allow Dr. Montie continue her studies on the role of acetylation of AR and to determine if blocking this process does reduce the death of cells with mutant KD
2007 - Two $ 25,000 research grants were awarded:
Two grants just funded by the KDA both attempt to investigate mechanisms to prevent the accumulation of the toxic fragment in cells containing the mutant AR.
Briefly, KD is caused by a genetic mutation to the gene that codes for the Androgen Receptor (AR) protein. This protein mediates all the actions of the androgen hormones testosterone and dihydrotestosterone, DHT. In the cells of normal males, the AR is found in the cytoplasm of the cell. Upon the addition of an androgen hormone (either testosterone or DHT), the hormone binds to the AR and the hormone/AR complex travels to the nucleus of the cell where it initiates the masculine changes that are associated with the presence of androgens (beard growth, for example). If there is no androgen present, then the AR never enters the nucleus and there are no changes – this is essentially what occurs in females. Since women do not possess androgens, the AR does nothing in cells and there are no masculine effects. The AR in the nucleus is ultimately destroyed by a cell structure known as the proteasome. In individuals with KD, the cell is unable to completely destroy the AR that enters the nucleus - but it can destroy the AR that does not enter the nucleus and this inadequate digestion apparently results in the production of a fragment of the mutant AR that is toxic to the cells – thus the cells die and this leads to the formation of the symptoms of KD. This appears to explain why women carriers do not show major symptoms. Since the levels of androgens in women are low, the mutant AR does not enter the nucleus and the cell does not create the toxic fragment.
A $25,000 grant was awarded to Maria Pennuto, Ph.D. from the National Institute of Health. Dr. Pennuto has spent the past few years investigating the molecular switches on the AR that are involved in the movement of the AR into the nucleus upon addition of hormone. She has discovered that certain chemical changes to the AR seem to reduce the ability of the AR to bind to hormone and thus not enter the nucleus (and cause KD!!). She has discovered that the exposure of cells to a substance known as IGF-1 can induce these chemical changes to occur to the mutant AR and thus prevent the movement of the AR to the nucleus. Thus, the addition of IGF-1 to a cell with mutant AR appears to prevent the formation of the toxic fragment and thus the cell stays alive. Dr. Pennuto will continue this work by determining if any other chemical changes to the AR may alter its movement to the nucleus and she will also determine if IGF-1 prevents the formation of KD symptoms in a KD mice model (up to this time, the effect of IGF-1 has only been shown to work in cell cultures. This work could lead to new therapies for KD.
Another $ 25,000 grant was awarded to Udai Bhan Pandey, Ph.D. from the University of Pennsylvania. The proposal by Dr. Pandey and Dr. Paul Taylor continues the work that they did (in part thanks to a previous KDA grant!). They previously reported that KD symptoms in a fly model of KD could be reduced by activating another mechanism for destroying the KD in the nucleus, by passing the need for the proteasome. This alternate pathway, known as autophagy, apparently is capable of destroying the toxic fragment. They did this by making the fly over produce another protein known as HDAC6. By doing this, they were able to demonstrate that the overproduction of HDAC6 did not show cell death despite the presence of the mutant KD. They will now try to continue this work as they attempt to find other proteins that may affect this activity of HDAC6 to stimulate autophagy and thus help prevent the cell death associated with KD.
2006 - Two $ 25,000 research grants were awarded:
A $25,000 grant was awarded to Chawnshang Chang Ph.D. from the University of Rochester. His research plans to develop a treatment regimen for Kennedy’s Disease targeting the poly Q-expanded mutant AR. This concept may be a way to cure the disease.
Another $25,000 grant was awarded to Udai Bhan Pandey Ph.D. from the University of Pennsylvania. Dr. Pandey proposes to use molecular genetic approaches in Drosophila to characterize the mechanism of suppression by HDAC6. His long-term goal is to contribute to the development of therapeutic interventions for Kennedy’s Disease.
2005 - One $ 25,000 research grant was awarded:
A $25,000 emergency funding grant was awarded to J. Paul Taylor, MD, Ph.D. from the University of Pennsylvania. The grant helped support Dr. Taylor and his team's research using the Drosophila melanogaster (fruitfly) model system to investigate the molecular pathogenesis of Spinal and Bulbar Muscular Atrophy (aka Kennedy's Disease). In response to the KDA's grant, we received the following email from Dr. Taylor referencing the status of his current Kennedy's Disease research. "... This (grant) could be a life saver. We have made great strides with our work, in fact, we have a manuscript on our Kennedy's Disease work that has received good reviews. This work was largely funded by (the) KDA and I have been anxiously waiting for this work to be accepted for publication before alerting you. I have also had two graduate students join my lab who are doing their Ph.D. thesis work on Kennedy's Disease.
2004 - One $ 25,000 research grant was awarded:
A $25,000 grant was awarded to Andrew Lieberman, MD, Ph.D., University of Michigan for development of First Ever Kennedy's Disease Knock-In Mouse Model. "Thanks so much for all of your hard work on our behalf! It sure is inspirational for us to know that the KDA cares enough about this project to launch a difficult fund raising drive to support our work, and it's really gratifying to see that the generosity of the KDA membership made these efforts successful so quickly." -- Dr. Andrew Lieberman
2003 - One $ 25,000 research grant was awarded:
A $25,000 grant was awarded to J. Paul Taylor, MD, PhD, University of Pennsylvania for developing the Drosophila melanogaster (fruitfly) model system to investigate the molecular pathogenesis of Spinal and Bulbar Muscular Atrophy.