2013 Report of AKC-CHF Grants 2017-10-04T18:47:23+00:00

2013 Report of AKC-CHF Grants

01925-A: Discovery of Novel Micro-Ribonucleic Acids for Diagnosis and Prognosis of Canine Hemangiosarcoma
Grant Status: Open
Grant Amount: $12,960
Dr. Bruce F Smith, VMD PhD, Auburn University
April 1, 2013 – March 31, 2014
Sponsor(s): American German Shepherd Dog Charitable Foundation, Inc
Breed(s): Golden Retriever
Disease(s): Hemangiosarcoma
Research Program Area: Oncology – Hemangiosarcoma
Abstract
Hemangiosarcoma is a tumor of blood vessels, often located in the spleen, causing death in almost 100% of patients. The prognosis remains poor even if aggressive treatments are undertaken. It is currently impossible to differentiate hemangiosarcoma from other masses of the spleen without the expense of surgery and biopsy. A special problem for dog owners is the fact that the main differential diagnosis is splenic hematoma, which is benign and carries a good prognosis. Micro-ribonucleic acids (miRNAs) are small ribonucleic acids (RNAs) that prevent messenger RNAs (mRNAs) from creating proteins. MiRNAs have been linked with cancer and can act as tumor suppressors (prevent cell overgrowth), oncogenes (promote cell overgrowth), or both depending on the tumor type. Dr. Smith will profile miRNAs in dogs with hemangiosarcoma using RNA sequencing and compare these to expression profiles of other canine splenic masses and normal spleens of dogs. The end result will be discovery of novel biological markers of hemangiosarcoma to enhance diagnosis and prognosis in dogs.

01905-A: Identification of Enhanced Methods for Semen Preservation
Grant Status: Open
Grant Amount: $12,776.94
Dr. Margaret V. Root Kustritz, DVM PhD, University of Minnesota
February 1, 2013 – March 31, 2013
Sponsor(s): American German Shepherd Dog Charitable Foundation, Inc
Breed(s): -All Dogs
Research Program Area: Reproductive Conditions and Disease
Abstract
Semen is not sterile and cannot be collected without bacterial contamination due to presence of normal bacterial flora on the urethral mucosa. Bacterial growth in stored semen is presumed to be controlled by addition of an extender to the semen, a liquid medium containing nutrients, buffering agents, and antibiotics. There is evidence in large animal species of varying efficacy of antibiotics in controlling bacterial growth in extended semen, raising concerns about passage of disease through semen. To date there are no studies in the dog documenting control of growth of aerobic and anaerobic bacteria, and mycoplasma through use of antibiotics in commercially available canine semen extenders. Dr. Kustritz will evaluate whether growth of aerobic, anaerobic and mycoplasma species will be controlled in semen extended with commercial canine extender when held at refrigerator (5?C) or room (20?C) temperatures for up to 48 hours. The results will provide practical information to breeders who ship and receive semen.

1843: Further Investigation of the Genes Controlling Canine Leukemia to Properly Diagnose and Control the Disease
Grant Status: Open
Grant Amount: $131,265
Dr. Matthew Breen, PhD, North Carolina State University
January 1, 2013 – December 31, 2014
Sponsor(s): American German Shepherd Dog Charitable Foundation, Inc., Golden Retriever Foundation, Poodle Club of America Foundation, Rottweiler Health Foundation
Breed(s): -All Dogs
Research Program Area: Oncology
Abstract
Leukemia represents a range of cancers, most often classified according to the type of blood cell affected and the clinical progression. Leukemia may be chronic, progressing slowly for many years with minimal symptoms, or acute, with sudden onset and rapid progression of symptoms, often resulting in euthanasia. The true incidence of leukemia in dogs is unknown, but consensus opinion is that many cases remain undiagnosed. In previous studies Dr. Breen found that canine leukemia presents with characteristic chromosomal and genetic changes shared with those known in human leukemia. In humans these chromosomal and genetic aberrations have been linked to disease progression and response to therapeutics, and in turn, this information drives clinical management of the patient. In this multicenter study, Dr. Breen’s group will use high-resolution genome-wide chromosomal evaluation to screen a large cohort of canine leukemia patients for the presence of recurrent chromosomal and genetic changes. This study will enhance our understanding of the pathogenesis of canine leukemia by identifying regions of the canine genome, and thus individual genes that may be critical for the control of these cancers. Additionally, this study will provide data that will impact our knowledge of the corresponding human disease.

1787: Clinical Advancement of a Cancer Vaccine in Dogs
Grant Status: Open
Grant Amount: $96,660
Dr. Nicola J Mason, BVetMed, PhD, University of Pennsylvania
January 1, 2013 – December 31, 2014
Sponsor(s): American Belgian Tervuren Club, Inc., American German Shepherd Dog Charitable Foundation, Inc., American Spaniel Club Foundation, Australian Shepherd Health & Genetics Institute, Canaan Dog Club of America, Cardigan Welsh Corgi Club of America, Clumber Spaniel Health Foundation, Flat-Coated Retriever Foundation, Golden Retriever Foundation, Great Dane Club of America, Irish Wolfhound Foundation, Newfoundland Club of America Charitable Trust, Old English Sheepdog Club of America, Pembroke Welsh Corgi Club of America, Portuguese Water Dog Foundation, Puli Club of America, Inc., United States Australian Shepherd Foundation
Breed(s): Akita, American Foxhound, Basset Hound, Bloodhound, Border Collie, Boxer, Bulldog, Dandie Dinmont Terrier, Doberman Pinscher, Golden Retriever, Irish Water Spaniel, Kerry Blue Terrier, Labrador Retriever, Newfoundland, Rottweiler, Sealyham Terrier, Tibetan Terrier
Disease(s): Lymphoma
Research Program Area: Oncology – Lymphoma
Abstract
Canine lymphoma is the most common blood-based cancer in dogs with an estimated annual incidence of 30/100,000. Chemotherapy induces remission in 75-85% of patients; however, the majority of patients relapse with drug-resistant lymphoma within 8-10 months of diagnosis and most dogs die of their disease shortly thereafter. Cell-based vaccine strategies that stimulate anti-tumor immunity have shown promise in the treatment of many different cancer types including non-Hodgkin’s lymphoma (NHL) in humans. In a previous study Dr. Mason developed a cell-based vaccine to induce anti-tumor immunity in dogs with NHL. Initial studies were hopeful as this early vaccine significantly prolonged second remission duration and overall survival, but ultimately the vaccine did not prevent relapse. These early findings suggest that while the lymphoma vaccine stimulated anti-tumor immunity it will require immunological boosting to achieve prolonged cancer-free survival. In the current study, Dr. Mason will optimize her cell-based vaccine approach to induce functional, long lasting tumor-specific immune responses that will prevent relapse and prolong survival in dogs with NHL.

1771: Defining the Unique Genetic Markers in Dogs That Define Immune Function, Disease Resistance and Tissue Transplantation
Grant Status: Open
Grant Amount: $178,200
Dr. Aravind Ramakrishnan, MD, Fred Hutchinson Cancer Research Center
January 1, 2013 – December 31, 2014
Sponsor(s): Old English Sheepdog Club of America, American German Shepherd Dog Charitable Foundation, Inc.,
Breed(s): -All Dogs
Disease(s): Major Histocompatibility Complex
Research Program Area: Immunology and Infectious Disease
Abstract
The Major Histocompatibility Complex (MHC) genes encode proteins that are critical for a wide range of biological functions, from immune protection against infectious disease to the predisposition of an individual to develop diabetes and auto immune diseases. The MHC genes in the dog are incompletely characterized, thereby severely limiting our ability to full define the cause of many canine diseases. Dr. Ramakrishnan has developed improved methods for identifying the different forms of canine MHC genes in a large number of dogs of diverse breeds. In this study he will characterize the patterns of MHC genetic variation in over 1200 dogs from at least 50 breeds using a high throughput sequencing strategy. The distribution and frequency of different forms of each of these genes and their specific clustering among different breeds will greatly enhance our knowledge of the genetic diversity among breeds. The methodology and data gained from this study will enhance the power of association studies between MHC types and canine diseases. Such a database will also enable tissue transplantation from unrelated but matched donors as a treatment for advanced malignancies (stem cell transplants) and other diseases (tissue transplantation). Fully defining the canine MHC will have broad impact across canine health, including oncology, immunology and infectious disease.

1759: Disrupting the Differentiation of Cancer Stem Cells to Prevent the Spread of Hemangiosarcoma
Grant Status: Open
Grant Amount: $233,914
Dr. Jaime F Modiano, VMD PhD, University of Minnesota
January 1, 2013 – December 31, 2015
Sponsor(s): American Belgian Tervuren Club, Inc., American German Shepherd Dog Charitable Foundation, Inc., American Spaniel Club Foundation, Australian Shepherd Health & Genetics Institute, Australian Terrier Club of America, Briard Club of America Health & Education Trust, Dachshund Club of America, Inc., Flat-Coated Retriever Foundation, Golden Retriever Foundation, Great Dane Club of America, Ibizan Hound Club of the United States, Irish Setter Club of America Foundation, Italian Greyhound Club of America, Keeshond Club of America, Keeshond Donors Circle Trust, Leonberger Health Foundation, Norwegian Lundehund Association of America, Inc., Old English Sheepdog Club of America, Poodle Club of America Foundation, Portuguese Water Dog Foundation, Rottweiler Health Foundation, Saluki Health Research, Inc., Skye Terrier Club of America, Starlight Fund, TarTan Gordon Setter Club, University of Minnesota – DAF for EIC Royalties, Versatility in Poodles, Inc., Welsh Springer Spaniel Club of America, White Shepherd Genetics Project
Breed(s): Airedale Terrier, Australian Shepherd, Belgian Sheepdog, Bernese Mountain Dog, Bouvier des Flandres, Boxer, Flat-Coated Retriever, German Shepherd Dog, German Shorthaired Pointer, Keeshond, Labrador Retriever, Portuguese Water Dog, Saluki, Siberian Husky
Disease(s): Hemangiosarcoma
Research Program Area: Oncology – Hemangiosarcoma
Abstract
Hemangiosarcoma is a rapidly fatal disease. The lifetime risk is alarmingly high for some breeds like Golden Retrievers (~20% will die of this disease) and Portuguese Water Dogs (~15% will die of this disease). The risk of hemangiosarcoma is not limited to just these breeds but is considered a research priority for 40 different breed Parent Clubs. Despite considerable efforts to find effective treatments, the outcome for dogs with hemangiosarcoma has changed very little over the past 30 years. Recent evidence suggests hemangiosarcoma conforms to the “cancer stem cell” model, where a defined subset of cells is responsible for initiating and maintaining the tumor. These cells are resistant to conventional therapies and are very adaptable, being able to survive in a variety of tissues in the body. For this project, Dr. Modiano proposes to reduce the malignant potential of hemangiosarcoma stem cells by forcing them to terminally differentiate into cells which can no longer self-renew. He further proposes that by disrupting their ability to self-renew he will enhance the sensitivity of these cells to conventional and targeted therapies and improve the outcomes of dogs with this disease.

01726-A: Identifying Growth Factors That Promote the Spread of Osteosarcoma
Grant Status: Open
Grant Amount: $12,960
Dr. Stuart Helfand, DVM, Oregon State University
April 1, 2012 – September 30, 2013
Sponsor(s): American Bullmastiff Association, German Shepherd Dog Club of America, Irish Setter Club of America Foundation
Breed(s): Irish Wolfhound
Disease(s): Osteosarcoma
Research Program Area: Oncology – Osteosarcoma
Abstract
Canine osteosarcoma (OSA) is the most common primary bone tumor in dogs, comprising 85% of all bone malignancies and affecting up to 10,000 dogs in the Unites States annually and is almost always fatal due to spread (metastasis). In an effort to develop novel therapeutics to circumvent the progression of this disease, key factors regulating tumor cell invasion and metastasis to surrounding tissues must be identified. Hepatocyte growth factor (HGF) is one such factor as is it is known to contribute to motility, migration, and invasion by canine and human OSA tumor cells. The recent introduction of tyrosine kinase inhibitors (TKIs), drugs that target growth pathways in cells such as that sparked by HGF, offer new opportunities to improve therapy for canine OSA. The purpose of this study is to evaluate how HGF contributes to canine OSA cell migration and invasion and begin to explore the potential to block HGF effects by switching off this circuit with several TKIs that target different points in the HGF pathway. TKIs have been implicated in decreasing the adhesiveness and motility of various types of human cancer cells, which may serve to inhibit the spread of highly metastatic tumors such as canine OSA.

01708-A: Validation of a general method for enrichment of canine cancer stem cells
Grant Status: Open
Grant Amount: $12,960
Dr. Timothy D. O’Brien, DVM PhD, University of Minnesota
January 1, 2012 – December 31, 2012
Sponsor(s): American German Shepherd Dog Charitable Foundation, Inc German Shepherd Dog Club of America, Norwegian Elkhound Association of America, Inc., Siberian Husky Club of America
Breed(s): German Shepherd Dog
Research Program Area: Oncology
Abstract
Osteosarcoma, hemangiosarcoma, and glioblastoma multiforme are three types of incurable cancers that are responsible for reduced quality of life and significant mortality in dogs. The processes that control the clinical behavior of these tumors is not well understood, but recent research suggests that a specialized group of cells called “cancer stem cells” (or CSCs) might play important roles. CSCs are important in the development, growth and spread of various malignant tumors of humans, so it would not be surprising if they also contributed to canine cancers. However, the definitive existence of CSCs in canine cancers remains to be proven. Defining these cells and understanding their basic biology will be necessary to develop cures for cancer, so we developed a unique, widely applicable method to enrich for cells that behave like CSCs from canine tumors. This method allows us to make direct comparisons among CSCs from osteosarcoma, hemangiosarcoma, and glioblastoma to find unique or shared targets for prevention and therapy. The objective for this ACORN project is to show that the presumed CSCs are indeed uniquely capable of forming tumors using the gold standard assay of “xenotransplantation at limiting dilution”. This will confirm the utility of our method and provide critical support for further groundbreaking genetic analyses of CSCs that will help us reach the promise of targeted therapies to reduce incidence and improve outcomes for dogs with these deadly cancers.
Publication(s)
– Frantz AM (2013). Comparative and Molecular Approaches to Improve Identification, Classification, and Therapeutic Options in Cancer. Doctoral Dissertation Thesis, University of Minnesota – Kim JH, Anderson KL, Frantz AM, Graef AJ, Scott MC, Robinson

01609: Use of Probiotic to Reduce the Symptoms of Inflammatory Bowel Disease
Grant Status: Open
Grant Amount: $97,416
Dr. Albert E. Jergens, DVM, PhD, Iowa State University
January 1, 2012 – December 31, 2014
Sponsor(s): American Belgian Malinois Club, American German Shepherd Dog Charitable Foundation, Inc., American Whippet Club, Belgian Sheepdog Club of America, Inc., Briard Club of America Health & Education Trust, English Setter Association of America, Inc., Health & Rescue Foundation of the Petit Basset Griffon Vendeen Club of America, Irish Setter Club of America Foundation, Keeshond Anonymous Fund, National Beagle Club, Old English Sheepdog Club of America, Papillon Club of America, Samoyed Club of America Education & Research Foundation, Siberian Husky Club of America, Westie Foundation of America, Inc., Yorkshire Terrier Club of America
Breed(s): Basenji, Boxer, Chinook, French Bulldog, German Shepherd Dog, Soft Coated Wheaten Terrier
Disease(s): Inflammatory Bowel Disease
Research Program Area: Gastrointestinal Disease
Abstract
Idiopathic inflammatory bowel disease (IBD) is a common cause of chronic gastrointestinal disease in dogs. Accumulating evidence in human IBD and animal models suggests that imbalances in composition of the intestinal microbiota contribute to the pathogenesis of chronic intestinal inflammation. Recent studies have also shown that dogs with IBD have distinctly different duodenal microbial communities compared to healthy dogs. Current treatments for IBD include the administration of nonspecific anti-inflammatory drugs which may confer serious side effects and do not address the underlying basis for disease, namely, altered microbial composition. Use of probiotics (viable, non-pathogenic bacteria that exert health benefits beyond basic nutrition) offers an attractive, physiologic, and non-toxic alternative to shift the balance to protective species and treat IBD. The aim of the proposed study is to investigate the clinical, microbiologic, and anti-inflammatory effects of probiotic VSL#3 in the treatment of canine IBD. These studies will provide highly relevant insight into the anti-inflammatory effects of probiotics for treatment of human and canine IBD.
Publication(s)
Otoni, R. Atilmann, M. Garcia-Sancho, et al. Serologic and fecal markers in prediction of acute disease course in canine chronic enteropathies. J Vet Intern Med 2012; 26:768-769. Slovak et al. Inter- and intra-observer assessment in the endoscopic assessment of canine inflammatory bowel disease. J Vet Inten Med 2013; in press.

01585: Evaluating a Safer, Less Toxic Radiation Therapy in the Treatment of Lymphoma
Grant Status: Open
Grant Amount: $93,140
Dr. Michael Deveau, DVM, MS, Texas A&M Research Foundation
January 1, 2012 – December 31, 2013
Sponsor(s): American Foxhound Club, American German Shepherd Dog Charitable Foundation, Inc., American Spaniel Club Foundation, Clumber Spaniel Club of America, English Setter Association of America, Inc., Keeshond Anonymous Fund, Pembroke Welsh Corgi Club of America, Yorkshire Terrier Club of America Foundation, Inc.
Breed(s): Akita, American Foxhound, Bloodhound, Border Collie, Dandie Dinmont Terrier, Doberman Pinscher, Irish Water Spaniel, Kerry Blue Terrier, Newfoundland, Sealyham Terrier, Tibetan Terrier
Disease(s): Lymphoma
Research Program Area: Oncology – Lymphoma
Abstract
Lymphoma is one of the most common neoplasms in canine companion animals accounting for upwards of 25% of all canine cancer. Chemotherapy results in high remissions rates but poor overall survival even with aggressive therapy. Lymphoma is extremely radiosensitive; however, incorporating full and half-body radiation therapy results in harmful toxicity to health tissue. In human medicine, full and half-body radiation has been abandoned for advanced Involved-Field Radiotherapy (IFRT) techniques utilizing advanced radiotherapy systems designed to kill diseased tissue while sparing normal tissues. As the technology becomes available in veterinary medicine, this treatment capability will also become available; however, there are no studies in the veterinary literature specifically interrogating this strategy. While demonstrable benefit is the ultimate clinical endpoint, it is critical to ensure safe implementation of IFRT for use in canine patients. To test feasibility and safety, Dr. Deveau will conduct a phase I study in which patients with advanced stage lymphoma will be treated with IFRT. Patients will be subjected to rigorous evaluation at each treatment and at one month intervals for dose limiting toxicities and/or adverse events, with the goal being to determine whether IFRT is a viable option for treatment of lymphoma in dogs.

01497-A: Dog leukocyte antigen 88 typing in German Shepherd Dogs having pancreatic insufficiency
Grant Status: Closed
Grant Amount: $10,649
Dr. Leigh Anne Clark, PhD, Clemson University
July 1, 2010 – September 30, 2011
Sponsor(s): American German Shepherd Dog Charitable Foundation, Inc., Health & Rescue Foundation of the Petit Basset Griffon Vendeen Club of America, White Shepherd Genetics Project
Breed(s): German Shepherd Dog
Disease(s): Pancreatic Acinar Atrophy
Project Summary
Pancreatic insufficiency in German Shepherd Dogs is caused by an autoimmune reaction that selectively destroys the pancreatic cells responsible for secreting digestive enzymes. Although once thought to be a simple recessive trait, recent work has shown that both genetic and environmental factors likely contribute to the disease. An initial whole-genome screen to identify genetic markers linked to pancreatic insufficiency yielded significant results on chromosome 12. This chromosomal region harbors the dog leukocyte antigen (DLA) system, the canine equivalent of the major histocompatibility complex (MHC), which is important in autoimmune and infectious diseases. Expression studies have shown that one MHC gene, DLA-88, is over-expressed in dogs with pancreatic insufficiency. The objective of this work was to determine if alleles of the MHC gene, DLA-88, are genetic risk factors for exocrine pancreatic insufficiency (EPI) in the German Shepherd Dog (GSD). DLA-88 sequences were generated for 148 unrelated GSDs. Complete genotypes were determined for 132 dogs (69 cases and 63 controls). Statistical calculations were performed to assess differences in allele frequencies between cases and controls. Significant associations were found with three alleles. One allele confers an increased risk for EPI. Over 20% of EPI cases had at least one copy of this allele, whereas only one copy was observed in the control population. Two other alleles were found to confer protection against EPI. These data support a role for the immune system in the development of EPI in GDSs.

01480: Leptospirosis: An Emerging Health Concern for Field Trial and Hunting Dogs
Grant Status: Open
Grant Amount: $73,620
Dr. Janet Foley, DVM, PhD, University of California, Davis
January 1, 2011 – December 31, 2013
Sponsor(s): Basenji Club of America, Inc. & Basenji Health Endowment, Border Terrier Club of America, English Springer Spaniel Field Trial Association, German Shepherd Dog Club of America, Golden Retriever Foundation, Labrador Retriever Club, Miniature Pinscher Club of America, Inc., Norwegian Elkhound Association of America, Inc., Pembroke Welsh Corgi Club of America, Siberian Husky Club of America, Versatility in Poodles, Inc.
Breed(s): -All Dogs
Disease(s): Leptospirosis
Research Program Area: Immunology and Infectious Disease
Abstract
Leptospirosis is a disease of dogs and humans that can be fatal, occurs worldwide, and originates from wildlife. The bacteria causing disease has many different strains, each coming from a different wildlife reservoir. Although vaccination of dogs against canine leptospirosis is available, it is not currently considered a core vaccine that should be administered to all dogs in the United States. Further, novel and potentially emerging vaccine-resistant strains from wildlife have been detected recently, thus potentially leaving a critical mass of dogs at risk for disease. At greatest risk are those dogs that spend excessive time in close association with wildlife through field trials or hunting. To address this concern, Dr. Foley will determine risk factors for, and clinical characteristics of, modern leptospirosis in dogs by retrospective analysis of two large databases. She will generate a statistical risk model and use Global Positioning System (GPS)-based mapping to detect spatial clustering of cases; develop a reliable, sensitive diagnostic method to identify active infections and differentiate among strains of Leptospira interrogans. Additionally, the Foley research group will prospectively sample dogs in high risk areas in order to acquire isolates for future pathogenesis study and molecular genotyping, and aid in the eventual evaluation of acute active infections of individual dogs to allow for successful treatment. Results from this study will provide valuable information that will improve success of patient management, reduce risk of infection to other dogs and humans, and inform future surveillance and vaccine efforts to improve canine health.
Publication(s)
Spatial and Temporal Patterns of Leptospira interrogans seropositivity in dogs in Northern California. Authors: J. Hennebelle, J. E. Foley, T.E. Carpenter, J.E. Sykes.

01429: Mechanistic Relationship of IL-8 in Cell Proliferation and Survival of Canine Hemangiosarcoma
Grant Status: Open
Grant Amount: $100,000
Dr. Jaime F Modiano, VMD PhD, University of Minnesota
January 1, 2011 – June 30, 2013
Sponsor(s): American Bouvier des Flandres Club – Bouvier Health Foundation, American Bullmastiff Association, American German Shepherd Dog Charitable Foundation, Inc., American Miniature Schnauzer Club, Inc., Australian Shepherd Health & Genetics Institute, Australian Terrier International, Briard Club of America Health & Education Trust, Estate of Virginia Lyn Tarquinio, Flat-Coated Retriever Foundation, German Shorthaired Pointer Club of America, Golden Retriever Foundation, Hoffman Miniature Schnauzer Donor Advised Fund, Irish Wolfhound Club of America, Inc., Labrador Retriever Club, Portuguese Water Dog Foundation, Rhodesian Ridgeback Club of the United States, Saluki Health Research, Inc., Staffordshire Bull Terrier Club of America, Starlight Fund, Tibetan Terrier Club of America/Tibetan Terrier Health & Welfare Foundation, United States Australian Shepherd Foundation
Breed(s): Bernese Mountain Dog, Boxer, German Shepherd Dog, Golden Retriever, Labrador Retriever, Portuguese Water Dog
Disease(s): Hemangiosarcoma
Research Program Area: Oncology – Hemangiosarcoma
Abstract
New insights into the mechanisms that control tumor progression have provoked considerable interest in the interaction of cancer cells with their microenvironment. Specifically, a molecule called IL-8 that can support tumor growth and survival, also recruits inflammatory cells and promotes blood vessel formation in the local tumor environment, enhances resistance to therapy, and facilitates metastasis in various aggressive cancers. Hemangiosarcoma (HSA) is an incurable, highly metastatic cancer that occurs commonly in dogs. There is virtually nothing known about how tumor cells and the microenvironment interact with each other in HSA, and more specifically, a role for IL-8 has not been investigated. In a recent study funded by CHF grant 422, we showed upregulation of IL-8 was a consistent feature that distinguished HSA cells from non-malignant endothelial cells, suggesting IL-8 might play a significant role in this disease. For this project, we will characterize the direct effects of IL-8 on HSA cells, an essential first step in the process to establish if and how this pleotropic molecule modulates disease progression. Our results will begin to clarify the importance of IL-8 production by HSA cells, and provide the foundation for subsequent studies to define its role regulating interactions between HSA cells and their microenvironment.

01418: Harnessing a Dog’s Own Immune System to Kill Lymphoma Tumor Cells
Grant Status: Open
Grant Amount: $150,000
Dr. Heather M. Wilson, DVM, Texas A&M Research Foundation
January 1, 2011 – December 31, 2013
Sponsor(s): American Belgian Tervuren Club, Inc., American Bullmastiff Association, American Chinese Crested Club, American German Shepherd Dog Charitable Foundation, Inc., American Miniature Schnauzer Club, Inc., American Shetland Sheepdog Association, Basset Hound Club of America, Inc., Central New Jersey Hound Association, English Setter Association of America, Inc., English Springer Spaniel Field Trial Association, French Bulldog Club of America, German Wirehaired Pointer Club of America, Golden Retriever Foundation, National Beagle Club, Otterhound Club of America, Portuguese Podengo Pequenos of America, Inc., Samoyed Club of America Education & Research Foundation
Breed(s): -All Dogs
Disease(s): Lymphoma
Research Program Area: Oncology – Lymphoma
Abstract
Lymphoma is the most common malignancy of dogs representing up to 25% of diagnosed cancers. Dogs often develop an aggressive form of lymphoma that is rarely curable, with most unfortunately succumbing to disease within 12 months of diagnosis despite best-available chemotherapies. Dr. Wilson will develop a new treatment to re-train the dog’s own immune system to attack the most common type of canine lymphoma, B-cell lymphoma. In order to accomplish this they will obtain a small number of circulating white blood cells, called T cells, from the blood of affected dogs and insert a gene that will cause the T cell to express a receptor which recognizes the tumor “fingerprint”. After docking with the lymphoma, the T cell will be triggered to mount an immune response against the tumor cells with the specific fingerprint. This therapy could be used alone or in combination with chemotherapy. Their preliminary data demonstrate that it is possible to genetically modify T cells. Further, they have been able to successfully harvest and grow T cells in the laboratory and return them safely to the dog. These infused cells can be found in the blood and tumor weeks after infusion, showing that it is possible for these cells to survive in the dog. If successful this study will be the first to develop an “in-dog” T-cell therapy targeting a tumor that has historically thought to be untreatable.
Publication(s)
O’Connor CM, Sheppard S, Hartline CA, Huls H, Johnson M, Palla SL, Maiti S, Ma W, Davis RE, Craig S, Lee DA, Champlin R, Wilson H, Cooper LJ. Adoptive T-cell therapy improves treatment of canine non-Hodgkin lymphoma post chemotherapy. Sci Rep. 2012;2:249. doi: 10.1038/srep00249. Epub 2012 Feb 13.

01364-A: Heat shock protein 72 (HSP72) as a biomarker for acclimation to, and performance in high level physical activity in pure-bred military working dogs
Grant Status: Closed
Grant Amount: $12,000
Dr. Yaron Bruchim, D.V.M., Hebrew University of Jerusalem
November 1, 2009 – December 31, 2011
Sponsor(s): American Belgian Malinois Club, American German Shepherd Dog Charitable Foundation, Inc.
Breed(s): Belgian Malinois, German Shepherd Dog
Abstract
Heat stroke (HS) is a devastating condition in both humans and companion animals, with reported death rates of 5% and 50%, respectively. Classical (environmental) heat stroke occurs when an animal or person are exposed to high environmental temperatures, such as when it is inadvertently locked in an unventilated vehicle. Exertional heat stroke occurs when an animal or person performers strenuous exercise, especially under heat stress. The Israel Defense Force Military Working Dog Unit (IDFMWDU) contains a majority of pure-bred Belgian malinois dogs imported periodically from Western Europe, where the combination of temperature and humidity, collectively termed heat stress are significantly lower than in Israel. Upon arrival, these dogs engage in a gradual training period with progressively increasing physical exercise, during which they are exposed to heavy heat stress. Although all dogs have a similar age, are the same breed, and are trained under identical conditions, some develop HS, while others do not. It has been previously shown that individuals have a genetic susceptibility for developing HS. The ability to test for this genetic susceptibility may prove itself useful in the selection of both canines and soldiers for military jobs with high risk for heat stroke, and aid in the prevention of this condition. Heat shock protein 72 (HSP72) is a protein that is induced in the body under conditions of increased heat stress. The purpose of our study is to investigate whether this protein can be used as a predictor for individual susceptibility to develop heat stroke in military working dogs.

Grant: 01271A: Mapping of Additional Genes Associated with Canine Degenerative Myelopathy
Principal Investigator: Dr. Kerstin Lindblad-Toh, PhD
Research Institution: Broad Institute
Grant Amount: $95,866.00
Start Date: 1/1/2010 End Date: 3/31/2012
Progress Report: 30 month
Report Due: 4/30/2012 Report Received: 4/30/2012
Sponsor(s): American Boxer Charitable Foundation, American German Shepherd Dog Charitable Foundation, Inc., Estate of Virginia Lyn Tarquinio, Orthopedic Foundation for Animals, Pembroke Welsh Corgi Club of America
Breed(s): Boxer, Pembroke Welsh Corgi
Disease(s): Degenerative Myelopathy
Abstract
Background: Degenerative myelopathy (DM) is a fatal disease of the spinal cord causing progressive paralysis in mature dogs. In grant 821 these researchers performed genome-wide association mapping and identified an SOD gene mutation which is present in many breeds. A DNA test can now identify dogs at risk of developing DM, although many dogs that have the mutation never develop clinical symptoms. The availability of the DM test has raised questions that must be answered before the test can be used by dog breeders. Objective: The researchers aim to find out if there genetic modifiers that could explain why some dogs with the mutation develop DM at 8 years of age while other 15 year-old dogs with the same mutation remain symptom free and if there are other mutations in the SOD1 gene or elsewhere that can cause DM or DM-like-diseases. The identification of modifier genes could prove particularly useful in breeds like the Boxer and Pembroke Welsh Corgi where the primary mutation is common, because it might provide an option fur selecting away from DM without disrupting breeding programs. Identification of additional genes will also give a better understanding of the disease and allow development of potential treatment strategies.
Report to Grant Sponsor from Investigator:
Degenerative myelopathy (DM) is a fatal disease of the spinal cord causing progressive paralysisin mature dogs. In grant 821, we performed genome-wide association mapping and identified a SOD1gene mutation, which is present in many breeds. A DNA test can now identify dogs at risk of developing DM, although many dogs that have the mutation never develop clinical symptoms. The availability of the DM test has raised questions that must be answered before this test can be optimally used by dog breeders. These include: Are there genetic modifiers that could explain why some dogs with the mutation develop DM at 8 years of age while other 15-year-old dogs with the same mutation remain symptom free? Are there other mutations in the SOD1 gene or elsewhere that can cause DM or DM-like diseases? We are making good progress on answering these questions, having preliminary data about modifier loci for both the Boxer and Pembroke Welsh Corgi breed that is being further evaluated. We have determined that the onset distribution is more tightly clustered in Boxers than in Pembroke Welsh Corgis (PWC), potentially making it harder to map modifier loci in Boxers than PWCs. The Boxer GWAs resulted only in two nonsignificant loci. Outside the scope of this grant we will now continue to collect samples for a modifier screen in a larger sample size. Three loci on chromosomes 25, 27 and 37 were significantly associated with ALS in PWC. The two loci on chromosomes 27 and 37 are well defined and each contains an excellent candidate gene expressed in the brain. All identified variants are currently being evaluated for functional potential. Once the most associated variants with the strongest association have been identified from the validation genotyping based on the targeted resequencing we will perform functional studies. We conclude that it is highly unlikely that cis-acting genetic modifiers contribute to the apparent differences in susceptibility to DM in the two Pembroke Welsh Corgis we studied. During the time-frame of this grant we also investigated histopathology of spinal cords and have now confirmed DM in over 20 purebred and mixed breed dogs. We identified a new SOD1 mutation in Bernese Mountain Dogs and currently are investigating a GSD with DM for a mutation in a different gene. Identification of additional genes will also give us a better understanding of the disease and allow development of potential treatment strategies.

01265: Understanding Mechanisms Involved in Canine Autoimmune and Inflammatory Disorders
Principal Investigator: Dr. Ronald Sluyter, Ph.D.
Research Institution: University of Wollongong

Grant Amount: $48,730.00
Start Date: 1/1/2010 End Date: June 30, 2012
Sponsor(s): American German Shepherd Dog Charitable Foundation, Inc., American Pointer Club, Collie Health Foundation, German Shepherd Dog Club of America, Health & Rescue Foundation of the Petit Basset Griffon Vendeen Club of America, National Beagle Club, Staffordshire Terrier Club of America
Abstract
Background: Autoimmune and inflammatory disorders have a major impact on the quality of life and lifespan of dogs. Studies in humans show an important role for an immune cell molecule (termed P2X7) in autoimmune and inflammatory disorders including inflammatory pain. P2X7 in these disorders has an ability to cause the release of key molecules (termed interleukins) that drive immunity and inflammation. Thus, P2X7 is attracting international interest as a therapeutic target in humans and currently undergoing clinical trails in patients with autoimmune or inflammatory disorders. The researchers have recently identified P2X7 on immune cells from dogs. Objective: The researchers will study P2X7 on canine immune cells, and its role in canine immunity and inflammation. They will also study the P2X7 gene in various dog breeds to determine if P2X7 differs between breeds and whether these differences contribute to disease susceptibility. New information about canine P2X7 will enable the use of currently available compounds and the development of other compounds to target P2X7, and treat autoimmune and inflammatory disorders, as well as inflammatory pain in dogs.
Report to Grant Sponsor from Investigator:
Autoimmune and inflammatory disorders have a major impact on the quality of life and
longevity of dogs. Studies in humans highlight an important role for a molecule (termed P2X7), present on white blood cells, in autoimmune and inflammatory disorders including inflammatory-related pain. The role of P2X7 in these disorders is largely attributed to its ability to cause the release of key molecules (termed interleukins) that drive immunity and inflammation. Thus, P2X7 is attracting considerable international interest as a therapeutic target in humans, with a number compounds, able to block P2X7, currently undergoing clinical trails in humans with autoimmune or inflammatory disorders. We have recently identified P2X7 on white blood cells from dogs (English Springer Spaniels). We propose to further study P2X7 on white blood cells from dogs, and its role in canine immunity and inflammation. We will also study the P2X7 gene in various dog breeds to determine if P2X7 differs between breeds and whether these differences contribute to disease susceptibility. New information about canine P2X7 will enable the use of currently available compounds and the development of other compounds to target P2X7, and treat autoimmune and inflammatory disorders, as well as inflammatory-related pain in dogs.
To date we have made an artificial form of P2X7 from white blood cells obtained from an English Springer Spaniel. This artificial P2X7 will serve as a useful tool to study the role of P2X7 in dogs. Information from these studies may be used to develop drugs that can block canine P2X7, and thereby potentially reduce inflammation and inflammatory-related pain in many dog disorders.
We have also confirmed the presence of P2X7 and found previously undetected immune molecules in dog white blood cells. These molecules may provide additional therapeutic targets in dogs. Moreover, we have found some of these molecules in dog kidney cells, which may be of therapeutic importance in canine kidney diseases.
Finally, we have begun measuring the relative amount of P2X7 in white blood cells from various dog breeds. Our data suggests that amounts of P2X7 differ between dog breeds. Moreover, we have collected genetic material from each of these dogs. Collectively, this information will be used to determine if genetic differences contribute to susceptibility to autoimmune and inflammatory disorders in various breeds.

01213-A: Determination of Outcome Measures for Clinical Progression and Morphometric Studies of Spinal Cord Disease in Degenerative Myelopathy Dogs
Grant Status: Closed
Grant Amount: $12,811
Dr. Joan R. Coates, DVM, University of Missouri, Columbia
January 1, 2009 – June 30, 2010
Sponsor(s): American Boxer Charitable Foundation
Breed(s): Boxer, Cardigan Welsh Corgi, Chesapeake Bay Retriever, German Shepherd Dog, Rhodesian Ridgeback
Disease(s): Degenerative Myelopathy
Project Summary
Degenerative myelopathy (DM) is a disease of the spinal cord causing progressive weakness in the hind limbs in adult dogs. Though most commonly reported in German Shepherds, high disease prevalence also exists in other dog breeds, such as Cardigan and Pembroke Welsh Corgis (PWC), Rhodesian Ridgebacks, and Boxer dogs. Genome-wide association mapping and a candidate gene approach identified a mutation in the canine SOD1 gene. Homozygosity for the mutant allele was associated with DM in five dog breeds which segregate for DM (Boxers, Pembroke Welsh Corgis, Rhodesian Ridgebacks, Chesapeake Bay Retrievers and German Shepherd dogs). Canine DM caused by SOD1 mutations resembles some forms of human ALS – Lou Gehrig’s disease. Better characterization of outcome assessment measures and understanding of the spinal cord pathology will assist in establishing collaborations with ALS researchers and in development of therapeutic drug trials similar to those of human ALS. The researchers hypothesize for this proposal that the spinal cord dysfunction and nerve root deterioration of canine DM will compare to that of human ALS. They are testing their hypothesis by performing clinical studies of gait and studies of spinal cord and nerve root tissues in DM affected dogs. The investigators were successful in obtaining matching funding from the ALS Association for completion of the proposed work. For the gait studies, they have digitized previously recorded gaits from DM affected dogs that have been serially evaluated over time. They continue to follow 2 DM-affected dogs and thus, will need to rely on archived cases recorded from 31 DM-affected dogs. The PI has one of her residents assigned to the project who will organize the blinded gait scoring using a validated gait scale. Thus far in total, they have 152 spinal cords submitted to the University of Missouri Veterinary Medical Diagnostic Laboratory specifically with a presumptive diagnosis of DM of which 109 have been DM confirmed (26 Boxer DM confirmed) and homozygous for the SOD1:c.118A allele; one dog (Chesapeake Bay Retriever) has been confirmed for DM and was A/G heterozygous. They have 44 spinal cords from various breeds (6 Boxers) submitted as control samples. They have decided to focus on three breeds to include the Pembroke Welsh Corgi, Bernese Mountain dog and Boxer because these are the breeds from which they have received the greatest number of samples. However, they continue to collect and preserve specimens from all breeds. It has been opted to focus on the lower thoracic spinal cord for counting of neurons, neurons and evaluation of the aggregates. The investigators have established measurement techniques to quantitate SOD-1 aggregates which accumulate in the neuron, the cell body of the neuron and nerve roots. They have refined the techniques using an image analysis software program to evaluate the nerve roots and motor neurons and are analyzing these samples. Final results from this study will provide them with a much better understanding of the pathology that underlies DM and will provide quantitative markers to measure the efficacy of therapeutic studies.

01212-A: Phenotypic Characterization of Peripheral Nerve Disease in Degenerative Myelopathy Dogs
Grant Status: Closed
Grant Amount: $12,092
Dr. Joan R. Coates, DVM, University of Missouri, Columbia
January 1, 2009 – June 30, 2010
Sponsor(s): American Boxer Charitable Foundation
Breed(s): Boxer, Cardigan Welsh Corgi, Chesapeake Bay Retriever, German Shepherd Dog, Rhodesian Ridgeback
Disease(s): Degenerative Myelopathy

Project Summary
Degenerative myelopathy (DM) is a disease of the spinal cord causing progressive weakness in the hind limbs which progresses to affect the front limbs and shows signs also in the muscles and nerves. Though most commonly reported in German Shepherds, high disease prevalence also exists in other breeds, such as Cardigan and Pembroke Welsh Corgis (PWC), Rhodesian Ridgebacks, and Boxer dogs. Genome-wide association mapping and a candidate gene approach identified a mutation in the canine SOD1 gene. Homozygosity for the mutant allele was associated with DM in five dog breeds which segregate for DM (Boxers, Pembroke Welsh Corgis, Rhodesian Ridgebacks, Chesapeake Bay Retrievers and German Shepherd dogs). Canine DM caused by SOD1 mutations resembles some forms of human ALS – Lou Gehrig’s disease. Better characterization of the clinical disease spectrum in the DM affected dog, especially the nerve and muscle pathology will assist in establishing collaborations with ALS researchers and in development of therapeutic drug trials similar to those of human ALS. We hypothesize for this proposal that the nerve and muscle deterioration of canine DM will compare to that of human ALS. We are testing our hypothesis by performing clinical studies of the nerve and muscle and studying the pathology in DM affected dogs. The investigators were successful in obtaining matching funding from the ALS Association for completion of the proposed work. We have had opportunities to perform electrodiagnostic testing on 4 dogs prior to necropsy and 3 dogs early in the disease process. However, we have established a motor unit number estimation technique in dogs that will have potential use to measure disease progression when testing therapies. This work was funded by a University of Missouri Clinician Scientist Award. We have prepared a kit to collect quality samples for nerve and muscle evaluations. In total, we have collected 74 muscle and nerve specimens from various breeds of DM affected dogs and 49 control dogs of older age. We have decided to focus on two breeds, the Pembroke Welsh Corgi and Boxer due to sample numbers because these are the breeds from which we have received the greatest number of samples. Morphometric analysis has been completed and data analyzed. There is clear evidence for peripheral nerve disease both qualitatively and quantitatively in both breeds. Findings and discussions will be included in detail in the paper under preparation.

01169-A: Identifying Mutations in Genes Associated with Canine Hemangiosarcoma: Denser Fine Mapping of the Associated Locus in Chromosome 26
Grant Status: Closed
Grant Amount: $12,466.9
Dr. Chieko Azuma, DVM PhD, Tufts University
May 1, 2008 – August 31, 2008
Breed(s): German Shepherd Dog, Golden Retriever, Labrador Retriever
Disease(s): Hemangiosarcoma

Abstract
Hemangiosarcoma (HSA), a malignant tumor of blood vessels, is a significant health concern in dogs, with a reported incidence of up to 2% of all tumors. HSA can affect all dogs, but a particularly high disease incidence has been reported in certain breeds, such as golden retriever (15%), German shepherd (10%), and Labrador retriever. The higher incidence in these particular breeds suggests that genetic risk factors exist. We have identified seven regions in the canine genome associated with HSA in golden retrievers using a newly developed powerful analytical method in order to search for small differences in the patterns of DNA. Subsequently, DNA patterns have been compared with five other breeds and all risk factors appear to be shared with at least one other breed. We now aim to perform denser fine mapping to validate and further narrow down the associated locus on chromosome 26, which is likely to contain a gene associated with hemangiosarcoma. Once the mutations have been identified and their presence in different breeds assessed, it will be possible to rapidly develop genetic tests for carriers of HSA. Ultimately, understanding of the disease biology will lead to prevention and better treatment of HSA.

01168-A: Identifying Mutations in Genes Associated with Canine Hemangiosarcoma:
Denser Fine Mapping of the Associated Locus in Chromosome 5
Grant Status: Closed
Grant Amount: $12,960
Dr. Chieko Azuma, DVM PhD, Tufts University
July 1, 2008 – April 30, 2009
Sponsor(s): LEAP Agility
Breed(s): German Shepherd Dog, Golden Retriever, Labrador Retriever
Disease(s): Hemangiosarcoma
Abstract
Hemangiosarcoma (HSA), a malignant tumor of blood vessels, is a significant health concern in dogs, with a reported incidence of up to 2% of all tumors. HSA can affect all dogs, but a particularly high disease incidence has been reported in certain breeds, such as golden retriever (15%), German shepherd (10%), and Labrador retriever. The higher incidence in these particular breeds suggests that genetic risk factors exist. We have identified seven regions in the canine genome associated with HSA in golden retrievers using a newly developed powerful analytical method in order to search for small differences in the patterns of DNA. Subsequently, DNA patterns have been compared with five other breeds and all risk factors appear to be shared with at least one other breed. We now aim to perform denser fine mapping to validate and further narrow down the associated locus on chromosome 5 which is the most likely to contain a gene associated with hemangiosarcoma. Once the mutations have been identified and their presence in different breeds assessed, it will be possible to rapidly develop genetic tests for carriers of HSA. Ultimately, understanding of the disease biology will lead to prevention and better treatment of HSA.

01147: Identifying Mutations in Genes Associated with Canine Hemangiosarcoma
Grant Status: Closed
Grant Amount: $74,332.47
Dr. Chieko Azuma, DVM PhD, Tufts University
January 1, 2009 – June 30, 2010
Sponsor(s): American Bullmastiff Association, American German Shepherd Dog Charitable Foundation, Inc., Briard Club of America Health & Education Trust, Clumber Spaniel Club of America, Delaware County Kennel Club, English Setter Association of America, Inc., Flat-Coated Retriever Foundation, French Bulldog Rescue League, Golden Retriever Foundation, Greyhound Club of America, Health & Rescue Foundation of the Petit Basset Griffon Vendeen Club of America, Irish Terrier Club of America, Labrador Retriever Club, Leonberger Health Foundation, National Amateur Retriever Club, Poodle Club of America Foundation, Saluki Health Research, Inc., Scottish Terrier Club of America, Siberian Husky Club of America, United States Australian Shepherd Foundation
Breed(s): German Shepherd Dog, Golden Retriever, Labrador Retriever
Disease(s): Hemangiosarcoma

Project Summary
Hemangiosarcoma (HSA), a malignant tumor of blood vessels, is a significant health concern in dogs, with a reported incidence of up to 2% of all tumors. HSA can affect all dogs, but a particularly high disease incidence has been reported in certain breeds, such as Golden Retriever (15%), German Shepherd Dog (10%), and Labrador Retriever, suggesting that genetic risk factors exist. We have identified six regions in the canine genome that differ in golden retrievers with and without HSA and verified the findings with more advanced technology in this project. We are currently identifying the actual mutations. So far no mutations in candidate genes have been found, supporting the major role of regulatory mutations. Once the mutation has been found, it will be possible to rapidly develop genetic tests for risk assessment for susceptibility of HSA. Interpretation of these DNA tests will require the consideration of markers for all genes simultaneously as well as assessing risks for different types of cancer. The presence of multiple interacting genes, some at high frequency in the population, will make it difficult to reduce the disease frequency quickly, but should still allow for informed breeding. In addition, by examining the frequency of these mutations in other breeds we can determine which other breeds need to be screened for these mutations and to what degree they contribute to the risk of HSA in specific breeds. Still, perhaps the most significant outcome of knowing the actual mutations is that it might suggest which dogs should be under surveillance or preventive care. The identification of actual mutations should also lead to further study of functional effects of the causative mutations thereby increasing the understanding of the disease mechanism. A better molecular understanding will suggest novel treatment options and possible new drug targets. Due to aggressive nature, HSA is uniquely qualified for studying local invasion, angiogenesis and metastasis, and developing therapeutic intervention in dogs and humans.

01131: Genetic Background and the Angiogenic Phenotype in Cancer
Grant Status: Open
Grant Amount: $254,871
Dr. Jaime F Modiano, VMD PhD, University of Minnesota
January 1, 2010 – December 31, 2012
Sponsor(s): Akita Club of America, Inc., American Boxer Charitable Foundation, American Brittany Club, American Bullmastiff Association, American German Shepherd Dog Charitable Foundation, Inc., American Spaniel Club Foundation, Australian Shepherd Health & Genetics Institute, Basset Hound Club of America, Inc., Bearded Collie Club of America, Clumber Spaniel Club of America, Fort Worth Kennel Club, French Bulldog Club of America, Golden Retriever Foundation, Irish Setter Club of America, Inc., Keeshond Club of America, Keeshond Donors Circle Trust, Laura J. Niles Foundation, Inc., Mastiff Club of America, National Beagle Club, Newfoundland Club of America Charitable Trust, Portuguese Water Dog Club of America, Inc., Portuguese Water Dog Foundation, Saluki Club of America, Inc., Saluki Health Research, Inc., Scottish Terrier Club of America, St. Bernard Club of America, Starlight Fund, United States Australian Shepherd Foundation, Vizsla Club of America Welfare Foundation
Breed(s): German Shepherd Dog, Golden Retriever, Portuguese Water Dog
Disease(s): Hemangiosarcoma

Abstract
Background: Certain dog breeds are prone to develop certain types of cancer; yet, there has been little progress to define genes or other factors that account for this risk. The researchers’ recent work on hemangiosarcoma is the first to clearly demonstrate that a dog’s genetic background, defined by “breed,” can influence the type of genes that show up as tumors. This means that certain breeds are diagnosed with specific cancers more frequently than others because of the behavior of tumors after they show up, and not simply because they show up more frequently. Specifically, this may apply to the observed tendency for hemangiosarcoma seen in Golden Retrievers, German Shepherd Dogs, and Portuguese Water Dogs. In addition, one-size-fits-all therapies may be not enough to effectively treat this disease. Objective: This project will continue the researchers’ observations on gene appearance profiles in hemangiosarcoma from Golden Retrievers to German Shepherd Dogs and Portuguese Water Dogs, and it also will define how new targeted therapies may effectively control the disease in these and other dog breeds.

01012-A: Genetic Studies in Chronic Superficial Keratitis in German Shepherd Dogs
Grant Status: Closed
Grant Amount: $12,960
Dr. Guillermo Giovambattista,, National University of La Plata
August 1, 2007 – July 31, 2008
Sponsor(s): American German Shepherd Dog Charitable Foundation, Inc.
Breed(s): German Shepherd Dog
Disease(s): Chronic Superficial Keratitis
Abstract
Chronic superficial keratitis (CSK) is a progressive, inflammatory, and potentially blinding disease of the canine bilateral cornea. It is also known as German Shepherd Pannus, Uberreitier?s syndrome, and degenerative Pannus. The German Sherpherd breed is most commonly affected. The diffuse pattern of stain deposition and the absence of staining of specific epithelial structures indicated that CSK is not a classical autoimmune disease similar to any disease as the pemphigus group or to the systemic lupus erythematosus. Although the results may implicate CSK as an immune-mediated disease, nonspecific factors could be ruled out. The normal central cornea has little MHC class II expression, aberrant expression occurs in CSK, associated with secretion of gamma interferon by infiltrating CD4-expressing lymphocytes. Although this change is likely to be a secondary feature of the CSK lesion, increased MHC class II expression may play a part in perpetuating the corneal inflammation seen in the disease. Given the previously reported association between the CSK and up-regulation of major histocompatability class II antigen expression, the primary goal of this project is to determine the association between the dog leukocyte antigen system (DLA) and the disease in German Shepherd dog. This will allow to detect individuals susceptible to develop the disease before the clinical signs appeared.

00934: Investigation of Inherited Diseases of the German Shepherd Dog Using the SNP Array
Grant Status: Closed
Grant Amount: $139,775
Dr. Leigh Anne Clark, PhD, Texas A&M University
May 1, 2008 – October 31, 2010
Sponsor(s): American German Shepherd Dog Charitable Foundation, Inc., Estate of Virginia Lyn Tarquinio, German Shepherd Dog Club of America, White Shepherd Genetics Project
Breed(s): German Shepherd Dog
Disease(s): Pancreatic Acinar Atrophy, Degenerative Myelopathy, Megaesophagus

Project Summary
The German Shepherd Dog (GSD) is a very popular pet in the United States (third most registered breed in 2006). According to the Canine Inherited Disorders Database, the breed is afflicted with approximately 50 hereditary disorders, including degenerative myelopathy (DM), pancreatic acinar atrophy (PAA), and megaesophagus (ME). These diseases disrupt the quality of life for both dog and owner. There are no means to identify carrier or affected dogs before the onset of clinical signs, making it difficult to eliminate these diseases from the breed. The experiments for this project were designed to scan the entire genome for markers associated with several disease traits simultaneously. The three diseases focused on in this study were pancreatic acinar atrophy (PAA), megaesophagus (ME), and degenerative myelopathy (DM). An array containing over 127,000 single nucleotide polymorphisms (SNPs) was probed with 153 DNA samples from either healthy German Shepherds (GSDs) or those affected with PAA, ME or DM. Results for DM confirm a mutation recently described by another research group on chromosome 31 (SOD1), and identified a new location that may harbor a modifier mutation. ME results indicate that the disease is not inherited in an autosomal recessive fashion. A region on chromosome 12 may harbor mutations contributing to ME, but requires further investigation. Results for PAA highlight the extreme complexity of the disease. A region on chromosome 12 near the canine major histocompatibility complex appears to be associated with the disease and is currently the focus of another study.

00871-A: Canine Anal Furunculosis
Grant Status: Closed
Grant Amount: $12,960
Dr. Lorna J. Kennedy, PhD, University of Manchester
December 1, 2006 – December 30, 2008
Sponsor(s): German Shepherd Dog Club of America
Breed(s): German Shepherd Dog
Disease(s): Anal Furunculosis
Project Summary
We have previously identified a gene associated with Anal Furunculosis (AF) in a region of the genome that is rich in immune genes. We have now performed a whole genome scan comparing 24 German Shepherd dogs having AF with 36 unaffected control dogs, which has revealed 46 SNPs that are significantly associated with AF, and are spread over 18 different canine chromosomes. Four of these SNP associations are highly significant and give us great hope that we will be able to identify other genes/loci associated with this disease. If we can identify such genes, it may be possible to start selective breeding to reduce the risk of German Shepherd dogs developing AF.

00870-A: Analysis of Pancreatic Acinar Atrophy in the German Shepherd Dog Using the SNP Array
Grant Status: Closed
Grant Amount: $12,960
Dr. Leigh Anne Clark, PhD, Texas A&M University
February 1, 2007 – July 31, 2008
Sponsor(s): American German Shepherd Dog Charitable Foundation, Inc.
Breed(s): German Shepherd Dog
Disease(s): Pancreatic Acinar Atrophy
Abstract
Pancreatic acinar atrophy (PAA) is a digestive disorder observed primarily in the German Shepherd Dog (GSD). Presently, there are no methods to diagnose the disease prior to the onset of clinical symptoms, which include maldigestion, weight loss, and loose stools. Diagnosis of EPI is achieved by measurement of serum canine trypsin-like immunoreactivity (cTLI), with severely decreased concentrations being diagnostic for PAA. Although symptoms can be controlled with supplemental enzymes, treatment is costly and not always effective. It is unclear if PAA is controlled by one gene or multiple loci and previous studies have failed to elucidate the causative gene(s). We propose herein to use a novel approach, the canine SNP array, to evaluate differences between affected and unaffected GSDs in an effort to identify regions of the genome that may harbor genes implicated in PAA.

00760: Cellular Genomics – Molecular Cytogenetic Investigation of Canine Soft Tissue Sarcomas
Grant Status: Closed
Grant Amount: $135,963
Dr. Matthew Breen, PhD, North Carolina State University
October 1, 2007 – September 30, 2009
Sponsor(s): Australian Shepherd Health & Genetics Institute, Berner Lovers, Bernese Mountain Dog Club of America, Briard Club of America Health & Education Trust, Briard Health Alliance, Flat-Coated Retriever Foundation, Golden Retriever Foundation, Starlight Fund
Breed(s): Australian Shepherd, Bernese Mountain Dog, Flat-Coated Retriever, German Shepherd Dog, Golden Retriever
Disease(s): Soft-tissue Sarcoma

Project Summary
Histiocytic malignancies, often referred to as histiocytic sarcoma (HS) and/or malignant histiocytosis (MH) are of major health concern to a variety of breeds, including Bernese Mountain Dog and Flat coated Retriever. In this project we aimed to recruit and analyze 75 cases of MH/HS from these two breeds, using genome-wide array based comparative genomic hybridization (aCGH), multicolour fluorescence in situ hybridization (FISH) analysis and loss of heterozygosity (LOH) analysis. Importantly, this study included BMDs from both the USA and from France. As such we were able to determine that there are no significant differences the genome wide aberrations profiles of either population, suggesting that the Berner populations of the USA and Europe may be considered as a single large population for the purpose of genetic investigations. Due to additional resources becoming available, we were able to evaluate a total of 133 cases of MH/HS from BMDs and FCRs, and identify numerous recurrent DNA copy number that are shared between these breeds and unique to each breed, The regions of the genome recurrently involved have been interrogated and contain several key genes that are known to be associated with the cancer process. The data in this study revealed several key findings: 1) MH/HS from BMDs resident in the USA and in France have recurrent DNA copy number changes that do not differ significantly, suggesting that the strong association between breed and disease type is not affected by the gene pool of the breed. This indicates that we may be able to consider the BMDs in USA and Europe as a single population. Importantly, as we develop new treatments for this devastating cancer, they likely will apply to all BMDs with MH/HS, regardless of geographic origin. 2) Analysis of histiocytic malignancies from BMDs and FCRs indicates that, in general, there is a gross level of shared genetic changes, with 23 regions across both breeds being recurrent in >30% of the cases. The five most frequently occurring shared regions were highly recurrent in each breed (present in >50% of the cases) and so these data suggest that there is a strong association between genomic change and histiocytic malignancies. 3) At least three of these five highly recurrent shared regions contain genes that are known to be associated with cancers. 4) This study detected 13 aberrant region of the canine genome (on seven different chromosomes) that are significantly associated with the presence of HS affecting either internal organs or just a limb. This association also separated breed (BMD ‘vs’ FCR) and so it is not possible to determine if there is a correlation between 1) breed and cytogenetic profile, or between 2) anatomical location of the MH tumors internal masses and cytogenetic profiles, since there is such a strong association between BMDs and the presence of internal masses. To resolve this we would need to investigate the cytogenetic profiles of other breeds that present with internal MH masses. Further analysis would allow us to determine if it mass location or breed that drives the cytogenetic profiles. 5) Analysis of 25 cases of hemangiosarcoma in a range of breeds, using 1Mb resolution aCGH analysis, revealed the presence of highly chaotic genome reorganization. Many of the chromosome changes present are shared with other cancers, but we are unable to form any concrete conclusions about breed association or disease association without further case analysis. The next phase of this investigation will be to evaluate the status of the genes we have identified to be in regions of significance, both in affected and unaffected individuals

00593B: Mapping Genes Associated with Canine Hemangiosarcoma
Grant Status: Closed
Grant Amount: $96,908
Dr. Chieko Azuma, DVM PhD, Tufts University
April 1, 2006 – September 30, 2008
Breed(s): German Shepherd Dog, Golden Retriever, Labrador Retriever
Disease(s): Hemangiosarcoma
Project Summary
Hemangiosarcoma (HSA), a malignant tumor of vascular endothelial cells, is a significant health concern in dogs, with an incidence of ~2% of all tumors. A national heath survey of golden retriever reported that neoplasia accounted for >60% of all reported deaths and HAS was the most common malignant tumor affecting >15% of golden retrievers. A particularly high disease incidence of hemangiosarcoma in golden retriever suggests a genetic susceptibility. The purpose of this study is to identify the mutations causing the increased risk for hemangiosarcoma in golden retriever. To do this, we have proposed to compare the genotypes of dogs diagnosed with HSA with healthy older dogs using a statistical analysis. To date, we have collected 125 blood samples from golden retrievers diagnosed with HSA and more than 400 healthy golden retrievers over 8 years old. We have identified six regions in chromosomes associated with HSA and are have narrowed these to precise regions (a few hundred thousand base). We now need to find the precise mutations that cause the disease and link them to functionality. In the long term, this work should allow the development of specific genetic tests for carriers of HSA. Ultimately, understanding of the disease biology, which will lead to identification of target genes for prevention, early detection and novel treatments of this malignancy.

00454: Analysis of a Candidate Gene for Pancreatic Acinar Atrophy in the German Shepherd Dog
Grant Status: Closed
Grant Amount: $22,680
Dr. Keith E. Murphy, PhD, Texas A&M University
October 1, 2004 – September 30, 2005
Sponsor(s): American German Shepherd Dog Charitable Foundation, Inc.
Breed(s): German Shepherd Dog
Disease(s): Pancreatic Acinar Atrophy
Abstract
Although pancreatic acinar atrophy is primarily a disease found in the German Shepherd Dog, it is also diagnosed in other breeds such as the Rough Collie and Dachshund. Diagnosis is usually not made until later in life, therefore, affected and carrier dogs are unknowingly bred. When diagnosed, the current 3treatment is enzyme therapy, which has proven effective in managing the disease. However, this treatment can be costly and many owners cannot afford this form of therapy and may be faced with the decision to euthanize their pet. If a causative mutation is found, a genetic test will be developed to determine the affection status of dogs at a very early age, and breeding practices can be altered so that the disease is not passed on to further generations.

00276: Microarray Analysis for Cardiac Gene Expression in German Shepherd Dogs with Sudden Death
Grant Status: Closed
Grant Amount: $108,648
Dr. Sydney N. Moise, DVM, Cornell University
October 1, 2004 – September 30, 2007
Sponsor(s): American German Shepherd Dog Charitable Foundation, Inc., German Shepherd Dog Club of America
Breed(s): German Shepherd Dog
Abstract
Sudden death in dogs due to cardiac disease is common. Many of these dogs die because of abnormal beats of the heart. Such diseases are likely inherited in the dog. Breeds at risk for such a cardiac death include the Doberman Pinscher, Boxer, Great Dane, Irish Wolfhound, Afgan Hound, Newfoundland and the German Shepherd Dog (GSD). To discover the genetic mutations that cause the diseases in each of these breeds will take many years of work. Research to identify the means of marking abnormal genes will help us to find the mutations. Even though the mutations will be different, research in one breed will help other breeds as we can use the gained information as we search the canine genome for the fatal inherited mistake. GSD have an inherited propensity for sudden death due to the occurrence of abnormal heart beats that develop primarily during sleep or with certain anesthetic agents. We have the needed pedigrees from GSD for our research. In this proposal we put forth a comprehensive plan to used stored samples from GSD that have died and their relatives so that we can identify potential abnormal genes and develop the markers for the most likely genes that could be involved in the disorder.

0002277: Identification of the Genetic Cause or Causes for Cataracts in Several Breeds
Grant Status: Closed
Grant Amount: $70,000
Dr. George J. Brewer, MD, University of Michigan
January 1, 2002 – December 31, 2004
Sponsor(s): American Miniature Schnauzer Club, Inc., Golden Retriever Foundation, Labrador Retriever Club, Samoyed Club of America Education & Research Foundation, San Joaquin Kennel Club, Siberian Husky Club of America, Westie Foundation of America, Inc.
Breed(s): Boston Terrier, Cocker Spaniel, German Shepherd Dog, Golden Retriever, Labrador Retriever, Miniature Poodle, Miniature Schnauzer, Standard Poodle, Toy Poodle, West Highland White Terrier
Disease(s): Cataracts
Abstract
Inherited cataract is a major health problem in dogs, with a significant frequency in 66 breeds. In some breeds the puppy is born with cataract (congenital) while in others it develops at six months of age, and in others at 12-18 months, long after the animal has been placed. Obviously, inherited cataract places a large emotional and financial burden on the dog fancy. As the first step in our attach on inherited canine cataract, we will look for linkage between a cataract gene and a DNA marker in nine breeds, selected because they show clinical variability in cataract type. Choosing multiple types of cataracts in various breeds increases our chances of multiple hits with our list of candidate genes. The nine breeds are Labrador Retrievers, Golden Retrievers, Cocker Spaniels, Poodles, German Shepherds, Miniature Schnauzers, Boston Terriers and West Highland White Terriers. Other breeds will also be helped because th discovered causative mutations will likely be shared by many other breeds. As a backup approach, we’ll screen “”pre-hoc”” candidate genes for causality to the extent that time permits. In the end we will have developed DNA tests for many if not most of the inherited canine cataracts.

1628: The Genetics of Hypertrophic Osteodystrophy in the Weimaraner
Grant Status: Closed
Grant Amount: $10,000
Dr. John Angles, BVSc, University of California, Davis
October 26, 1998 – October 25, 1999
Sponsor(s): American German Shepherd Dog Charitable Foundation, Inc., Weimaraner Club of America
Breed(s): German Shepherd Dog, Weimaraner
Disease(s): Hypertrophic Osteodystrophy
Project Summary
This study’s data indicate that Hypertrophic Osteodystrophy (HOD) in the Weimaraner is an autosomal recessive disease (affected pups receive a copy of the mutated gene from each parent). The researchers collected DNA samples from extended families, which should provide the groundwork for the discovery of a marker that will allow for HOD carrier detection and the eventual elimination of this disease from the breed. HOD is a common disease in rapidly growing, large and giant purebred dogs. The Weimaraner has an unusual form of the disease typically seen between 8 to 16 weeks of age, with severe systemic manifestations, including fever and multiple body organ inflammation. Unlike most breeds, these dogs often require treatment with prednisone to prevent death. Using survey data, the researchers in this study noted a strong association between HOD and recent vaccination, with over 70 percent of affected Weimaraners vaccinated in the preceding 3 to 5 days.

1633: Identification of a Genetic Marker for Pancreatic Acinar Atrophy Causing Exocrine Pancreatic Insufficiency in the German Shepherd Dog
Grant Status: Closed
Grant Amount: $50,771
Dr. David A Williams, PhD, Texas A&M University
August 2, 1999 – September 30, 2002
Sponsor(s): German Shepherd Dog Club of America
Breed(s): German Shepherd Dog
Disease(s): Pancreatic Acinar Atrophy
Abstract
Exocrine pancreatic insufficiency (EPI), a disease marked by chronic diarrhea and weight loss, is most commonly seen in German Shepherd Dogs. This condition is almost exclusively caused by atrophy of pancreatic cells that secrete digestive enzymes. This disease process is termed pancreatic acinar atrophy (PAA). Previous research has indicated that PAA is inherited in the German Shepherd Dog population of Finland. While the development of a serum assay for the major pancreatic digestive enzyme, canine trypsin-like immunoreactivity (cTLI), has led to a more accurate diagnosis of this condition, serum cTLI concentrations in affected dogs are normal prior to the development of PAA. Therefore, it has not been possible to take measures to decrease the incidence of this disorder. This project was designed to establish a pedigree of German Shepherd Dogs with EPI caused by PAA, confirm the inheritance of PAA, identify a marker for PAA in the genetic code, and develop an assay for this genetic marker. Results from this research will allow the screening of German Shepherd Dogs intended for breeding for carrier status for this disease and may lead to the eradication of this disease in the German Shepherd Dog breed.

1632: Canine Degenerative Myelopathy: Role of Methionine Supplementation and Further Evaluation of Associated Enteropathy
Grant Status: Closed
Grant Amount: $72,920
Dr. Joan R. Coates, DVM, Texas A&M University
October 1, 1998 – September 30, 2000
Sponsor(s): American German Shepherd Dog Charitable Foundation, Inc.
Breed(s): German Shepherd Dog
Disease(s): Degenerative Myelopathy
Abstract
Degenerative myelopathy (DM) is a degenerative spinal cord disease affecting primarily, but not exclusively, older German Shepherd Dogs. Affected dogs show progressive rear limb weakness and eventually paralysis. Intestinal abnormalities have been described in affected dogs, and we suspect that an essential amino acid, methionine, may be deficient secondary to abnormal intestinal absorption. An imbalance of methionine or related substances in the blood of spinal fluid may affect the production of myelin, the fatty covering of the nerves that is essential to their function. If this is true, dietary supplementation with methionine may halt neurologic deterioration or improve neurologic signs. Testing blood or spinal fluid for concentrations of methionine and related substances may allow dogs at risk to receive early treatment before clinical signs of disease become apparent. Identification of such dogs also could help prevent them from being used for breeding purposes. Newly established tests of intestinal function will be used to further characterize abnormalities in dogs with DM. We also will redefine neurologic-based testing procedures to monitor disease progression and will utilize modern methods to evaluate the microscopic characteristics of the disease. We anticipate that the results of the proposed studies will help assist in the accurate diagnosis and monitoring of DM, allowing evaluation of new therapies and assisting in the future identification of a genetic marker for DM.