We continue to be the world leader in funding diverse and groundbreaking research projects devoted to GNE Myopathy. Our team of new scientific investigators are growing at an impressive rate, and their dedication to data-sharing and genuine collaboration is keeping with our grant awards and proving to be of great value. Please see see our list of past and present grantees here.
We are always on the lookout for promising research projects that will directly impact the lives of GNE Myopathy patients. If you are a researcher interested in applying for funding from us, please see our Grant Application process.
2023 Awarded Grants
Dr. Julie M. Crudele
University of Washington, Department of Neurology
Codon optimization of GNE transgene as a method to reduce doses for GNEM AAV gene therapy
Treating GNE myopathy (GNEM) with gene therapy would require high doses of adeno-associated virus (AAV) to be effective, and that comes with risks. Therefore, GNEM gene therapy, and many other gene therapy efforts, would benefit from reducing the amount of AAV required for effective treatments. In this project, Dr. Crudele will use a codon optimization approach to reduce the doses of AAV needed to achieve the same level of efficacy. Codon optimization is a technique wherein a computer algorithm determines the DNA sequence that will lead to the highest levels of gene therapy expression. She will also improve the gene therapy approach by targeting DNA sequences known as CpGs to minimize the risk of adverse immune responses following gene therapy by removing as many CpGs as possible. Finally, this project will also test new AAVs that deliver gene therapies to skeletal muscles with very high efficiency. These experiments will test these new therapies in mice to determine which version leads to the highest expression of GNE and therefore would allow for the lowest, safest vector dose in humans.
Dr. Monkol Lek
Department of Genetics, Yale University School of Medicine
Supporting the International Gene Therapy Development Program
This project in the Lek lab is vital to support the NDF as they work on their flagship International Gene Therapy Development Program (IGTDP) to produce gene therapy for treatment of GNEM. This project will support the collaboration of leading gene therapy and GNEM experts to develop and begin FDA approved clinical trials for GNEM using AAV-mediated gene replacement therapy. The project will complete two critical work pieces of work necessary to develop a gene therapy. The first will determine the minimum dose needed to achieve GNE expression and functional improvement using a GNE mutant mouse. The second will establish a potency assay to measure AAV effectiveness, which is critical for validation across production batches. This work should support submission of a pre-IND package to the FDA by demonstrating the ability of the gene therapy to fix a mouse model which harbors GNE mutations. Once the IND is approved by the FDA, enabling the start of human clinical trials, this will be one of the world’s first gene therapies for GNEM.
Dr. Paul T. Martin
Abigail Wexner Research Institute, Nationwide Children’s Hospital
Studies of an inducible GNE knockout mouse model for GNE myopathy
The goal of this project is to test a new mouse model for GNE myopathy (GNEM). The mouse models for GNEM that are currently available have several problems that make them difficult to use, particularly when testing the efficacy of new therapies. Deletion of the mouse Gne gene from conception does not allow for mice to be born. Thus, Gne is an essential gene required for normal development, making it challenging to use such a mouse to study GNEM in adult muscle. By creating a mouse model where the mouse Gne gene can be induced to be deleted after development has been completed, we can better understand the mechanism by which this disease occurs, and, importantly, it will provide a more robust disease model for testing therapies. An improved mouse model of GNE myopathy is needed to move forward with gene therapy treatments for patients. Such a model will allow for a clear demonstration of therapeutic efficacy, and this type of data is needed before any therapy is given to a human patient.
Dr. Stella Mitrani-Rosenbaum
Goldyne Savad
GNE Myopathy: GneKO models for assessment of AAVGNE gene therapy
The main obstacle preventing the establishment of treatments for GNE myopathy (GNEM) is the lack of animal models where the efficacy of any potential treatment should be evaluated. Dr. Mitrani-Rosenbaum has spent the last few years on developing animal models to that purpose. Since Gne is essential in the entire organism for embryonic development and very early life, this project will develop and characterize three mice models in parallel: one which have no Gne protein only in muscles; the second one with no Gne in both muscles and liver (since liver is the main producer of sialic acid and could be a source of it to muscles); a third model where Gne is absent in the entire organism, but only after birth and weaning of the pups. We anticipate that at least one of these models will develop muscle pathology that could be targeted for treatment evaluation, in particular gene therapy.
2022 Awarded Grants
Dr. Stella Mitrani-Rosenbaum
Goldyne Savad Institute of Gene Therapy, Hadassah Hebrew University Medical Center
GNE Myopathy: Establishment of GNEM KO muscle cells for the assessment of GNE related biomarkers
GNE Myopathy is a neuromuscular adult onset disease characterized by slowly progressive muscle weakness, caused by recessive mutations in the GNE gene. Although the function of this protein is well known, the mechanism of the disease is still not clear. To date, no reliable animal model is available for the disease, and cellular models from GNE Myopathy patients’ muscle cells (expressing the mutated protein) are less informative than expected. Therefore, we reason that models where the effect of the GNE mutation could be more dramatic may be useful to determine additional function (s) of GNE specifically in muscle. In a system where the GNE protein is totally absent there is a better chance to see changes in the processes related to the GNE gene. We propose to establish cellular models of mouse and human muscle cells that completely lack GNE expression. For this purpose we will use the Sol8 mouse muscle lineage, and two primary genuine muscle cells derived from GNE Myopathy patients muscle biopsies from our unique cell bank. By the Crispr/Cas9 methodology we will knock out the GNE gene. We will then determine the changes at various levels in order to recognize the biological and molecular pathways affected in the GNE knocked out cells and define specific biomarkers. These cells could then be treated by AAVGNE gene therapy to evaluate its potential benefits in restoring the normal affected features.
2021 Awarded Grants
Dr. Rüdiger Horstkorte & Dr. Kaya Bork
Institute for Physiological Chemistry, Martin-Luther-Universität Halle–Wittenberg
Development of a sensitive and robust enzyme assays for GNE (UDP-N-acetylmannosamine 2-epimerase/N-acetylmannosamine kinase)
Glycosylation represents an important post-translation modification that occurs on proteins and lipids and contributes to their structural and functional properties. The outermost end of most glycan chains on (glyco)proteins are occupied by sialic acids. Sialic acids are 9 carbon acidic sugars which strongly influence the cell-cell and cell-matrix interactions. The UDP-N-acetylglucosamine-2-epimerase/N-acetyl-mannosamine kinase (GNE) catalyzes the first two steps of the sialic acid biosynthesis. Both enzyme activities are located on one polypeptide, therefore GNE represents a bifunctional enzyme. The epimerase activity of GNE underlies a strong feedback inhibition by CMP-Neuraminic 5-acid (Neu5Ac), the end-product of the metabolic pathway. Lack of GNE is lethal.
Mutations in the GNE gene generates GNE variants, which are involved in the development of the GNE myopathy (formerly hereditary inclusion body myopathy, HIBM), which is accompanied with a progressive amyotrophia during ageing. Prominently, proximal and distal skeletal muscles are affected. After two or three progressive decades, the patients are tightly restricted in their mobility. Many GNE mutations lead to decreased enzyme activity and therefore clinical trials tried to bypass the GNE defect by supplementation of the GNE products ManNAc or N-acetylneuraminic acid. Unfortunately, these clinical trials failed so far. One bottleneck of all GNE myopathy research is the lack of feasible enzymatic assays. Therefore the aim of this application is to develop a fast, reliable and robust assay based on thin layer chromatography (TLC), which would be an important piece for the development for further therapies of GNE myopathy including gene therapy.
Dr. Kelly Crowe
Department of Biology, School of Behavioral & Natural Sciences Mount St. Joseph University
Evaluation of Lectin Staining Biomarkers in Pre-Clinical Models of GNEM
As a gene therapy is in development to treat GNE myopathy, the goal of this project is to develop ways to quickly and accurately measure the effects of this therapy. This requires we find a biological indicator called a biomarker; in our case, a biomarker will measure the changes in muscle sialic acid levels produced by a successful GNE myopathy gene therapy. While the ultimate goal of a clinical trial is to find improvement of muscle function, biomarkers are particularly important for the Food and Drug Administration (FDA) to approve an investigational new drug (IND) application because biomarkers can detect biological changes happening in the muscle even before improvement of function can be measured. Our previous research tested potential biomarkers in samples of human muscle to compare their ability to detect changes that would happen in a clinical trial. However, before a gene therapy can go to clinical trial, it has to be shown to be effective and safe in a mouse model to allow its use in humans. Because of this, it is critical to make sure that any biomarker that we develop can work in both mouse pre-clinical trials and human clinical trials. Our project proposes to compare the ability of our potential biomarkers to detect how GNE gene therapy changes sialic acid levels in the muscle of existing mouse models of GNE myopathy. We will also expand on our previous studies to confirm that these biomarkers also function in human muscle cells.
Dr. Julie Crudele
Department of Neurology, University of Washington
Development and preclinical assessment of AAV-GNE gene therapy
Adeno-associated viral vector (AAV)-based gene therapy is in development for several genetic muscular diseases, and one AAV gene therapy for a muscle disease has already been approved by the FDA. These therapies require large doses of AAV, which comes with safety concerns. This proposal aims to develop an AAV gene therapy for GNE myopathy, whereby the AAV vector is repurposed to deliver an unmutated version of the GNE gene to muscles, with a particular focus on reducing the dose of AAV required. To do that, we are developing a novel expression cassette that will lead to high levels of both GNE and glycoprotein sialylation in skeletal muscles throughout the body. By replacing the mutated enzyme (GNE) as well as the downstream products of that enzyme (glycoprotein sialylation), we aim to treat the underlying disease pathology with a safe and effective gene therapy. This project will specifically test three such gene therapies, the best of which will be tested in a dose-finding study in a future funding cycle. The data generated by this proposal and a subsequent dose-finding study can be used to support an Investigational New Drug application to the FDA in preparation for a GNE myopathy gene therapy clinical trial.
Dr. Monkol Lek
Department of Genetics, Yale School of Medicine
High throughput genetic screening assay for GNE
The complete function of the GNE gene in humans remains poorly understood, particularly with respect to how amino acid changes caused by missense mutations can lead to a body-wise muscle pathology. In this grant, we propose to design a biological assay to better understand the impact of GNE patient mutations on the gene’s function. This assay can be used to help classify patient mutations in GNE that are currently classified as ‘VUS’ (variants of uncertain significance); which would help towards providing a definitive genetic diagnosis to enable these patients to participate in future clinical trials (e.g. gene therapy).
Dr. Paul Martin
Center for Gene Therapy, Abigail Wexner Research Institute at Nationwide Children’s Hospital
Development and gene therapy testing of a new disease model for GNE myopathy
GNE myopathy is a muscle disease where a mutation in the GNE gene gives rise to progressive muscle weakness and loss that has a major impact on patients’ quality of life. Our laboratory is interested in developing gene therapies for the treatment of GNE myopathy. These treatments use Adeno Associated Virus (or AAV) as a DNA delivery device to place a normal copy of the GNE gene into cells, thereby providing normal GNE gene function back to the patient. This type of gene therapy could be used to treat all patients with GNE myopathy, regardless of their particular genetic mutation. Once given, the gene therapy delivered can last for decades, perhaps a lifetime, in the patient. The current grant proposal seeks to build and test gene therapy in a new mouse model of GNE myopathy where the mouse GNE gene can be deleted in the adult animal. The mouse GNE gene is essential for development, so only deletion of the gene in the adult will allow for animals to be born. Such a new disease model is required to help investigators define the therapeutic potential of gene therapies and also drug-based therapies for GNE myopathy. It is also needed to define the dose of therapy required to correct the disease. Current models of GNE myopathy have problems that make translational research for this disease difficult. Making and testing of this new model should greatly speed needed research for this disease.
Dr. Stella Mitrani-Rosenbaum
Goldyne Savad Institute of Gene Therapy, Hadassah Hebrew University Medical Center
GNE Myopathy: Establishment of Gne KO muscle cells for the assessment of GNE related biomarkers
GNE Myopathy is a neuromuscular adult onset disease characterized by slowly progressive muscle weakness, caused by recessive mutations in the GNE gene. Although the function of this protein is well known, the mechanism of the disease is still not clear. To date, no reliable animal model is available for the disease, and cellular models from GNE Myopathy patients’ muscle cells (expressing the mutated protein) are less informative than expected. Therefore, we reason that models where the effect of the GNE mutation could be more dramatic may be useful to determine additional function (s) of GNE specifically in muscle. In a system where the GNE protein is totally absent there is a better chance to see changes in the processes related to the GNE gene. We propose to establish cellular models of mouse and human muscle cells that completely lack GNE expression. For this purpose we will use the Sol8 mouse muscle lineage, and two primary genuine muscle cells derived from GNE Myopathy patients muscle biopsies from our unique cell bank. By the Crispr/Cas9 methodology we will knock out the GNE gene. We will then determine the changes at various levels in order to recognize the biological and molecular pathways affected in the GNE knocked out cells and define specific biomarkers. These cells could then be treated by AAVGNE gene therapy to evaluate its potential benefits in restoring the normal affected features.
Dr. Stella Mitrani-Rosenbaum
Goldyne Savad Institute of Gene Therapy, Hadassah Hebrew University Medical Center
GNE Myopathy: GneKO muscle conditional mouse for the assessment of AAVGne gene therapy
GNE Myopathy is a unique neuromuscular adult onset disease characterized by slowly progressive distal and proximal muscle weakness, caused by recessive mutations in GNE. Although the function of this protein is known to be a key enzyme in the biosynthesis of sialic acid, no clear definite explanation has been provided to account for the muscle atrophic pathology. The lack of animal models severely impair the comprehensive understanding of GNE function, and, more importantly, prevents the development of an efficient system to assess any kind of treatment, and in particular an AAV platform based GNE gene therapy. The goal of this study is to develop critical tools for this purpose. Since the lack of GNE in the entire organism results in the death of the embryo before it is born, we will develop a mouse where GNE can be knocked out only in muscle, so the mice can be born and we can see the effect of the lack of GNE in muscle. By the CRISPR/Cas9 genome editing method, we have already generated the first step towards this goal, and want to continue towards the second and final steps. Once established, these mice will be used as a system to rescue the muscle pathology by AAV-GNE gene therapy.
Dr. Wakako Yoshioka
Department of Neuromuscular Research, National Institute of Neuroscience, National Center of Neurology and Psychiatry
Activation of endogenous mutated GNE product by small compound
Developing drugs from the very beginning, especially for diseases with small number of patients, is extremely difficult. In contrast, the repositioning of approved drugs, whose safety information is already available, will allow us to bypass the slow drug developing process even for extremely rare diseases. The enzymatic activity of GNE is known to be activated by specific allosteric site mutations, raising a possibility that even the GNE with GNE- myopathy-causing mutations may possibly be activated by the modification of such allosteric sites. Our computationally predicted molecular model of GNE indicates the presence of a deep narrow cavity. Filling this cavity by a small compound is likely to inhibit the binding of CMP-NeuAc to the allosteric site, leading to the activation of the GNE.
In this study, we therefore aim to find approved small molecular agents that can fit the cavity and activate the GNE with either p.D207V in epimerase domain or p.V603L in kinase domain, which are the two most common mutations among Japanese GNE myopathy patients, by in silico drug screening based on binding affinity. We will then narrow down the candidates by measuring GNE enzyme activity and confirm the efficacy of drugs using patients’ cells including mutations frequent in other ethnic populations.
We propose the repositioning of approved drugs using in-silico approach is a fast, an effective and realistic method to treat GNE myopathy, which is the mission of NDF.
2020 Awarded Grants
Current Status of Therapy Development
- Generation and characterization of new animal models of GNE Myopathy.
- Identification and development of biomarkers of the disease.
- Identification and optimization of a therapeutic approach (AAV-mediated gene therapy; gene correction strategies; drug replacement/development therapy
- Patient registry
- Repositories of data and biological specimens
Dr. Stella Mitrani-Rosenbaum
Hadassah Medical Center
Development of new animal models for GNE myopathy
Dr. Monkol Lek & Dr. Angela Lek
Yale University – Lek Lab
Biobank
Dr. Monkol Lek & PerkinElmer
Whole Genome Sequencing (WGS)
Dr. Marjan Huizing
NHGRI-NIH
Identification of Biomarker
Dr. Angela Lek
CRISPR gene editing approaches to GNE
Dr. Noah Weisleder
Ohio State University
Compromised membrane repair as a potential pathologic mechanism in GNEM
Dr. Paul Martin
Nationwide Children’s Hospital
Development of FDA-compliant gene therapy assays for GNE myopathy
Dr. Ichizo Nishino & Dr. Wakako Yoshioka
NCNP
Activation of endogenous mutated GNE product by small compounds