GlycoScience Research Inc.
Executive Summary of ovine GM1 ganglioside Project
The proposed project is based on the need to develop a safe, plentiful, source of GM1 ganglioside for therapeutic use for clinical trials in Huntington’s disease, Parkinson’s disease, spinal cord injury and stroke. The literature contains numerous recent publications on the potential applications of GM1 ganglioside. The missing link is a safe, plentiful and sustainable natural source for ganglioside production. Prior developments in the GM1 ganglioside field were pioneered by Fidia Pharma Spa in the late 80’s and early 90’s. Their technology was based on GM1 production from bovine brain collected at slaughter plants throughout Europe. Fidia had developed the GM1 ganglioside platform for many potential applications and had phase II clinical trials in progress for spinal cord injury and Parkinson’s disease when bovine spongiform encephalopathy (BSE, Mad cow disease) was diagnosed in cattle in England. With the discovery of BSE, raw material from non-source verified animals of any species could not be reliably used for ganglioside production for pharmaceutical use. With the Fidia technology, normal bovine brain ganglioside yields are approximately 230 mgs of GM1 ganglioside and 1400 mgs of mixed ganglioside per Kg of tissue. GlycoScience Research has developed a new, verified, ovine (sheep) source for GM1 ganglioside production that will exceed these levels 30 to 40 fold, and provide needed raw materials for pharmaceutical applications.
Ganglioside Production from Ovine GM1 gangliosidosis sheep
GlycoScience Research (GSR) has developed a flock of sheep that are carriers for GM1 gangliosidosis. Ovine GM1 gangliosidosis is an autosomal recessive, lysosomal storage disease in sheep associated with a profound deficiency of the lysosomal enzyme acid beta-galactosidase. The stored products include GM1 ganglioside, asialo GM1 ganglioside and galactose-containing glycoproteins and long chain galactose containing oligosaccharides. Affected animals contain markedly elevated levels of GM1 ganglioside and asialo GM1 ganglioside in the brain, spinal cord, and lesser amounts in other tissues in the body. Since this disease is an autosomal recessive in inheritance, 1 out of every 4 lambs from carrier x carrier mating will be born with this disease.
The GSR flock is certified in the USDA Export Monitored Scrapie Certification Program and has operated as a closed, high health sheep flock for the past 20 years. A molecular genetic test has been developed to determine genotype status of each individual animal and aid in the selection of carrier animals for flock expansion. Tissues for ganglioside production are collected at slaughter at approximately 4 to 5 months of age and stored at -20°F until processed. Currently, the carcass is not allowed to enter the commercial food chain; however, significant levels of GM1 ganglioside have not been detected in muscle tissue. There is no reason to believe that with the demonstration of normal food safety and wholesomeness standards, the meat product from these lambs will not be allowed into normal marketing channels. GSR has facilities to process all lambs currently produced, however, it is anticipated that a dedicated facility will be required to collect tissues from a large numbers of animals, as would be needed for pharmaceutical production. Ovine GM1 ganglioside is currently produced from brain tissue supplied by GSR to Avanti Polar Lipids in Alabaster, Alabama. Ovine GM1 ganglioside is currently produced for research use only. Levels of GM1 ganglioside from brain are approximately 1.5 – 2.0 grams per brain. Levels in spinal cord and other tissues such as salivary gland, adrenal gland, liver, kidney spleen, intestinal mucosa and many others are significantly elevated and maximum yield per lambs is expected to approach 4 to 5 grams based on preliminary data. A more thorough analysis of all tissues will need to be performed. Optimal extraction procedures, purification methods, maximum yields and production efficiencies will have to be optimized for large-scale production. Avanti Polar Lipids has the capacity to develop necessary protocols for GMP manufacturing of ovine GM1 or provide necessary expertise if production was performed by a newly created entity. Additional GM1 ganglioside can be recovered by conversion of polysialogangliosides or asialo GM1 to GM1 ganglioside depending on costs of production. The overall goal will be to maximize the production of GM1 ganglioside from each affected lamb at a production cost that will maximize the affordability of a finished product.
GSR currently has a flock of 450 carrier females and approximately 50 normal females that are certified in the USDA Export Monitored Scrapie Certification Program. We would continue to select primarily for carrier females, but maintain normal animals for use as recipients for embryo transfer procedures and production of additional carrier animals. A number of carrier rams from a variety of genetic backgrounds are available to breed the GSR flock and to be used in the establishment of new flocks. It is the vision of GSR to maintain a flock of carrier animals and provide the ovine GM1 gangliosidosis genetics to cooperator sheep flocks that would be willing to produce animals for pharmaceutical use. Facilities are currently available to house the existing flock; however, additional facilities are in development for future expansion. This flock is managed as a high health, sustainable commercial sheep flock. Extensive development has taken place over the past 20 years and currently, procedures for production of GM1 gangliosidosis lambs for pharmaceutical production are in place.
GSR’s current emphasis is utilizing GM1 for the treatment of Huntington’s Disease (HD). Through collaboration with Dr. Steven Hersch, Harvard/Massachusetts General Hospital, ovine GM1 has been shown to decrease the mutant (disease-causing) huntingtin protein by 50%. In order to facilitate research progress, GSR is working on identifying partners to bring ovine GM1 ganglioside to the market as soon as possible. The undertaking will require further expansion of the GM1 genetics. Currently, seventeen cooperator flocks, incorporating an additional 5000 ewes, have signed agreements with GSR to contribute production of affected and carrier animals into an integrated animal production system, while additional sheep producers are eager to join. Tissue collection and processing will eventually be conducted in a facility similar to a modern abattoir. We anticipate that meat and pelts from this project will be eventually marketed through normal channels once the safety concerns of the USDA are addressed. Ganglioside production must be optimized to achieve maximum yields per affected lamb in order to be able to provide adequate product for the intended markets. While other groups have suggested that synthetic ganglioside would be a more palatable alternative to a natural product, the reality is that the technology, although developed over a decade ago, cannot produce the significant quantities of GM1 ganglioside that will be required for pharmaceutical use. It is the belief of GSR that unverified raw materials from any species will not be acceptable by the FDA for pharmaceutical production. Currently raw material from ovine GM1 affected lambs is available to produce adequate material to conduct preliminary research and subsequently start a clinical trial for Huntington’s disease.
GlycoScience Research was incorporated in 1998 in White, SD. This family-owned corporation was formed to develop a new sheep source for production of GM1 ganglioside for use in pharmaceutical applications including Parkinson’s disease and spinal cord injury. The management team of GSR currently consists of its founders, Larry and Sue Holler. Larry has a DVM degree from Kansas State University and a Ph.D. from Washington State University. He is currently employed as a diagnostic pathologist and animal reproductive disease specialist in the Veterinary and Biomedical Sciences Department at South Dakota State University. Sue Holler has a B.S. in Animal Science and an M.S. in Reproductive Physiology from Purdue University. She has been employed in research, teaching and diagnostic positions at University of Idaho, Washington State University, and South Dakota State University respectively. Together Larry and Sue manage the farm and research flock.