Abstract:
Hemangiosarcomas (HSA) is a common and fatal cancer in dogs for which there is no effective treatment. Despite surgery and intensive chemotherapy, the median survival time for dogs diagnosed with HSA is little more than six months. From our own studies on canine cancer, expression of the enzyme telomerase allows cancer cells to become immortal and has emerged as a central and near universal marker of cancer, making it a candidate target for novel therapies. In this study we will explore the value of telomerase inhibition to treat HSA using the novel mechanism of RNA interference (RNAi). Our hypothesis is that potent inhibition using this technique will inhibit the immortal phenotype of the cancer cells and cause them to die. However, it is possible that a combined approach, targeting two molecular pathways, may offer greater therapeutic benefit. Consequently, we will also explore the potential synergism of combining telomerase inhibition with an alternative inhibitor of a further mechanism involved in cancer (receptor tyrosine kinase inhibition) on targeting this highly malignant tumor. In this we will use in vitro cell culture techniques for initial inhibition studies followed by studies in a novel canine HSA model system to ascertain the potential clinical merit of this approach for dogs with HSA.

North Carolina State University (University)
Completed Grant No: 2214T: Identification of a 5-10Mb BAC Set as a Cytogenetic Resource and for the Development of an Ordered CGH Microarray for Cancer Studies in the Dog
Disease: Cancer
Sponsors: American Boxer Charitable Foundation, Nestle Purina PetCare Company
Researchers:Matthew Breen, PhD
Breeds: All (non-specified)
Funding: ABCF - $12, 781 CHF matching funding: $12,781; Nestle Purina Co. $50,124 Total: $75,686

Abstract:
The study of aberrant chromosome structure has significantly increased our understanding of the cause and progression of human cancers. Many cancers are common to both dogs and humans, in part reflecting the high degree of similarity in their genetic material and in their environmental exposure to carcinogens. The extent and identity of chromosome aberrations associated with canine cancers, however, remains largely unknown. This is primarily due to the difficulty in identifying dog chromosomes by conventional means alone. Comparative genomic hybridization (CGH) is a technique that allows a comprehensive analysis of chromosome aberrations present within tumors. We have developed this technique for application to dog cancers and are obtaining valuable information. However, chromosome-based CGH is labor intensive, has a limited resolution and requires detailed knowledge of dog chromosomes. In this study we aim to identify a set of large insert clones that are evenly spaced, at small intervals, along all dog chromosomes. These clones will be a very valuable resource for chromosome studies of dog cancers. The clones will be used to generate an ordered microarray to replace chromosome-based analyses. This will significantly and rapidly advance the study of canine cancer, leading to improved diagnosis and prognosis and thus health and welfare.

AMC Cancer Research Center (University)
Active Grant No: 2254A: Heritable and Sporadic Genetic Lesions in Canine Lymphoma and Osteosarcoma
Disease: Cancer
Sponsors: Akita Club of America, Inc., American Belgian Tervuren Club, Inc., American Bloodhound Club, American Boxer Charitable Foundation, Atlantic States Briard Club, Inc., Bernese Mountain Dog Club of America, Briard Club of America, Bull Terrier Welfare Foundation, Flat-Coated Retriever Foundation, German Wirehaired Pointer Club of America, Golden Retriever Foundation, Irish Wolfhound Club of America, Inc., Jeffrey Pepper, Mastiff Club of America, Medallion Rottweiler Club, Nestle Purina PetCare Company, Newfoundland Club of America Charitable Trust, Otterhound Club of America, Portuguese Water Dog Club of America, Inc., Rhodesian Ridgeback Club of the United States, San Joaquin Kennel Club
Researchers: Jaime Modiano, VMD, PhD
Breed(s): Akita, Belgian Tervuren, Flat-Coated Retriever, Golden Retriever, Mastiff, Rottweiler
Funding: ABCF - $7500 CHF matching funding: $7500 Breed Clubs, CHF matching, and Nestle Purina Co. - $454,798 Total: $469, 798

Abstract:
Lymphoma (cancer of lymph glands) and osteosarcoma (bone cancer) are two common cancers of dogs with remarkable breed predisposition. Lymphoma accounts for approximately 20 percent of all canine tumors, and > 80 percent of cancers originating from blood cells. Osteosarcoma is the most common bone tumor in dogs, accounting for 85 percent of skeletal cancers. All cancers have a genetic basis, and in effect, these conditions represent various diseases, each sharing one or few genetic abnormalities that contributes to overall risk and treatment response. However, a means does not exist to identify individuals at risk, or tumors that are likely to respond to conventional therapy. We have identified individual genes and larger regions within the genome that appear to be important in canine cancer. For this project, we propose to confirm the frequency and significance of these genetic anomalies in lymphoma and osteosarcoma of Golden Retrievers, Rottweilers, Irish Setters, and Bernese Mountain Dogs. This work will begin to determine which of these anomalies may be heritable and which may be sporadic, and pave the way to apply this knowledge for clinical benefits by providing potential targets for treatment, and tools to define individual risk to develop these types of cancer or produce cancer-prone progeny.

University of Wisconsisn (University) 
Active Grant No: 22101: Development of Anti-Canine Il-2Ra Antibodies Using CpG Oligodeoxynucleotide Vaccination
Disease: Cancer
Sponsors: American Boxer Charitable Foundation, Golden Retriever Foundation, Nestle Purina PetCare Company
Researcher: Stuart Helfand, DVM
Breed(s): All (non-specified), Golden Retriever
Funding: ABCF - $5500 CHF matching funding: $5500 Breed Clubs, CHF matching and Nestle Purina Co. - $53,979 Total: $64,979

Abstract:
Despite progress in treating canine lymphoma, most affected dogs eventually develop resistance to chemotherapy and succumb to their disease. In human medicine, the ¿ subunit of the interleukin-2 receptor (IL-2R¿) has been developed as a target for Immunotherapy of chemoresistant lymphoma patients. Intensively studied as a molecule present on normal lymphocytes that are activated, IL-2R¿ has recently been developed as a target on malignant lymphocytes that have been found to express this protein inappropriately. This has been accomplished using anti-IL-2R¿ monoclonal antibody (Mab) to deliver cellular toxins or radioisotopes directly to the cancer cells, a strategy that overcomes drug resistance. We would now like to develop such an antibody against the canine Il-2R¿ subunit that could benefit dogs with lymphoma. Our rationale for pursuing this approach stems from molecular data we generated indicating that a high percentage of canine lymphomas from Golden Retrievers and Rottweilers express Il-2R¿. Having synthesized canine recombinant (cr) IL-2R¿ using the gene sequence we cloned, we are now ready to produce and validate Mabs against it. To accomplish these goals, 1) we will employ a novel immunization strategy to prepare murine anti-canine IL-2R¿ Mabs, 2) develop a test using anti-canine IL-2R¿ Mabs for the detection of IL-2R¿ shed into the blood of Golden Retriever and Rottweiler lymphoma patients, and 3) screen lymphoma biopsies from Golden Retrievers and Rottweilers for IL-2R¿. Mabs generated to Canine IL-2R¿ will be a powerful tool in the diagnosis, prognosis, and treatment of canine lymphoma.

University of Wisconsin (University)
Active Grant No: 2629: Clinical and Immunological Outcomes in Dogs with Osteosarcoma Treated with Intratumoral Interleukin-12 Microspheres
Disease: Cancer
Sponsors: Akita Club of America, Inc., American Bloodhound Club, American Boxer Charitable Foundation, American Bullmastiff Association, American German Shepherd Dog Charitable Foundation, Borzoi Club of America, Flat-Coated Retriever Foundation, Golden Retriever Foundation, Great Pyrenees Club of Puget Sound, Irish Wolfhound Club of America, Inc., Irish Wolfhound Foundation, Jeffrey Pepper, Newfoundland Club of America Charitable Trust, Rottweiler Health Foundation, St. Bernard Club of America, Starlight Fund
Researchers: Stuart Helfand, DVM
Breeds: Akita, All (non-specified), Bullmastiff, Flat-Coated Retriever, German Shepherd Dog, Golden Retriever
Funding: Grant was Pending as of May 2005. ABCF Funding not currently available.

Abstract:
Appendicular osteosarcoma, or bone cancer of the limbs, is an important tumor in dogs representing nearly 10% of all canine cancers. Despite progress in treating canine osteosarcoma using a combination of limb amputation and chemotherapy, life expectancy is not usually extended by more than 6-10 months compared to amputation alone. Death is due to dissemination of cancer cells beyond the leg and it is estimated that the cancer has already spread in at least 95% of dogs when it is initially diagnosed. Any pet owner who has lived through this disease can attest to the heartbreak of the news that tumor masses, initially undetectable at the time of diagnosis, have been discovered on a chest X-ray. Novel treatment regimens are urgently needed to improve the lives of large breed dogs such as Golden, Labrador an other Retriever4s, Rottweilers, Irish Wolfhounds, Great Danes, German Shepherds and others that are at greatest risk for developing this cancer. Stimulating the immune system of dogs with cancer has been a goal of veterinary cancer researchers for more than 20 years and osteosarcoma is a tumor that has shown positive responses to some of these interventions. This research proposes to add a potent new form of immunostimulation to the standard treatment for canine osteosarcoma. This strategy uses a powerful stimulant of the immune system called interleukin-12 (IL-12) that has been shown to induce strong antitumor responses in experimental animal models. Stimulated by IL-12, immune cells tolerant of cancer can be triggered to kill cancer cells throughout the body, resulting in the generation of an army of deadly circulating killer immune cells specific for the cancer. What¿s more, this type of cell has long-term memory for the cancer, in much the same way vaccination results in long-term immunity to infectious disease. Our laboratory has shown that IL-12 enhances killing of osteosarcoma cells by immune cells from dogs with this cancer. Now, we propose that injection of IL-12 directly into limb osteosarcoma using a novel (microsphere) formulation resulting in slow IL-12 release within the tumor environment will promote active antitumor immunity in dogs with osteosarcoma and lengthen their survival time. A number of pertinent immunological questions will also be addressed.

University of California, Davis (University)
Active Grant No: 2465: Identification and Characterization of Genetic Mutations in Canine Mast Cell Tumors
Disease: Cancer
Sponsors: American Boxer Charitable Foundation, American Bullmastiff Association, Chinese Shar-Pei Charitable Trust, Collie Health Foundation, French Bulldog Club of America, Golden Retriever Foundation, Jeffrey Pepper, Rhodesian Ridgeback Club of the United States, Staffordshire Bull Terrier Club of America
Researchers: Cheryl London, DVM, PhD
Breeds: All (non-specified), Bullmastiff, French Bulldog, Golden Retriever
Funding: ABCF - $9800 CHF matching funding: $9800; Breed Clubs, CHF matching. - $45,714 Total: $65,314

Abstract:
The most common malignant tumor in dogs is the mast cell tumor (MCT, a form of skin cancer), occurring with an incidence of close to 20 percent in the canine population. MCTs range from relatively benign to extremely aggressive, leading to tumor spread and eventual death. Particular breeds of dog are at risk for the development of this tumor, indicating a role for genetic factors. We have previously identified mutations in the gene c-kit in 30-50 percent of dog MCTs. c-Kit plays a critical role in regulating the growth and function of normal mast cells, and as the mutations we discovered cause uncontrolled function of c-kit, it is likely they influence MCT development in dogs. This proposal will establish a prospective tumor registry of dog MCTs to be used for investigation of the true incidence of c-kit mutations within specific dog breeds. Moreover, the studies outlined in this grant will identify additional genetic mutations present in dog MCTs that can be used for the development of new targeted therapeutics. In summary, this work will provide a much more detailed understanding of dog MCTs, thereby building a framework for the development of new therapies and strategies for disease prevention.

University of California, Davis (University)
Active Grant No: 2646: Characterization of Receptor Tyrosine Kinase Dysfunction in Malignant Histiocytosis
Disease: Cancer
Sponsors: American Boxer Charitable Foundation, Berner Lovers, Bernese Mountain Dog Club of America, Flat-Coated Retriever Foundation, Golden Retriever Foundation
Researcher: Cheryl London, DVM, PhD
Breeds: Bernese Mountain Dog, Flat-Coated Retriever, Golden Retriever, Rottweiler
Funding: Grant was Pending as of May 2005. ABCF Funding not currently available.

Abstract:
Malignant histiocytosis (MH), while rare in people, occurs frequently in certain breeds of dogs including Rottweilers, Golden Retrievers, Flat-Coated Retrievers and Bernese Mountain Dogs. There is no effective therapy for this disease and nearly all patients die within two to four months of diagnosis. The prupose of this proposal is to evaluate MH tumor specimens for mutations in genes that may contribute to the development of this devastating cancer. The genes of interest are those that code for proteins known as growth factor receptors. These proteins are present on the surface of the cell and when stimulated by growth factors, signal into the cell promoting cell survival and growth. Dysregulation of growth factor receptors is a common mechanism through which normal cells undergo transformation into cancer cells. Significant research has been directed towards the development of inhibitors capable of blocking the function of dysregulated receptors. Recent success of this approach has been realized with the inhibitor Gleevec in the treatment of chronic myelogenous leukemia in people. The purpose of this proposal is to identify growth factor receptors that are dysregulated in MH to provide the foundation for future clinical application of growth factor receptor inhibitors in the treatment of MH.

North Carolina State University (University)
Active Grant No: 2667: Cellular Genomics - Molecular Cytogenetic Investigation of Canine Soft Tissue Sarcomas
Disease(s): Cancer
Sponsor(s): American Boxer Charitable Foundation, Berner Lovers, Bernese Mountain Dog Club of America, Canaan Dog Club of America, Collie Health Foundation, Flat-Coated Retriever Foundation, Golden Retriever Foundation
Researcher(s): Matthew Breen, PhD
Breed(s): Bernese Mountain Dog, Flat-Coated Retriever, Golden Retriever
Funding: Grant was Pending as of May 2005. ABCF Funding not currently available.

Abstract:
It has been established that non-random chromosome aberrations are characteristic of specific types of many different human cancers. The knowledge of such aberrations has identified areas of the human genome to be targeted for further research. In the dog the extent and identity of chromosome aberrations associated with specific cancers is still largely unknown. In certain breeds, such as the Flat-Coated Retriever and Bernese Mountain Dog, soft tissue sarcomas account for up to 50% of all malignant tumors and thus represent a serious health and welfare issue for those breeds. These tumors are difficult to classify by conventional means and so attention is required to develop alternative approaches. Human soft tissue sarcomas have been demonstrated to be associated with specific chromosomal aberrations that have been shown to have both diagnostic and prognostic significance. This proposal will make use of major recent advances in canine molecular cytogenetics to identify recurrent chromosome aberrations associated with canine soft tissue sarcomas, in particular those of histiocytic origin. This project will identify areas of the canine genome associated with such cancers for further investigation at the sub-chromosomal level.

Ohio State University (University)
Completed Grant No: 1428: Inheritance Patterns and Molecular Genetic Analysis of Doberman Pinschers and Boxer Dogs with Familial Dilated Cardiomyopathy
Disease: Heart Disease
Sponsors: American Boxer Charitable Foundation, Great Dane Club of America
Researcher: Kathryn Meurs, DVM, PhD, DACVIM
Breeds: Boxer, Doberman Pinscher
Funding: ABCF - $45,891 CHF matching - $20, 891; Total: $66,783

Abstract:
This study found that ventricular arrhythmias in Boxers are inherited as an autosomal dominant trait. Researchers found that asymptomatic Boxers have frequent ventricular premature complexes (VPCs), a specific type of irregular heartbeat that is a common form of ventricular arrhythmia. They determined that a two- to three-minute electrocardiogram (ECG) is a poorly sensitive indicator of VCPs, and instead recommend the use of a 24-hour ambulatory ECG (Holter monitor), even on asymptomatic Boxers. In studying Doberman Pinschers, they ruled out some potential candidate genes for study by demonstrating that two specific proteins and a specific gene that are abnormal in some humans with dilated cardiomyopathy (DCM)¿a primary heart muscle disorder that can cause the heart to beat erratically¿are normal in Dobermans with DCM. Linkage analyses for both Boxers and Dobermans continue.

Completed Grant No: 2002: Evaluation of the Clinical Outcome of Asymptomatic Adult Boxers with Ventricular Arrythmias Over a Four-Year Period
Diseases: Heart Disease
Sponsors: American Boxer Charitable Foundation
Researcher: Kathryn Meurs, DVM, PhD, DACVIM
Breeds: Boxer
Funding: ABCF - $45,099 CHF matching - $90,199; Total: $90, 199

Abstract:
Heart disease in the Boxer was initially documented in the 1980s and referred to as a Boxer cardiomyopathy. More recent studies have confirmed that this disease is inherited and is primarily characterized by disturbances in the cardiac electrical system, fainting episodes and sudden death. The inherited nature of the disease has led to increased interest in the screening of dogs for the disease by electrocardiogram, ultrasound and Holter monitoring prior to using them for breeding. However, the interpretation of the results is difficult, since many adult Boxers have some abnormalities detected on at least one of the tests and there is no available information regarding the relationship between these findings and the likelihood of development of clinical signs. The objective of this study is to evaluate the correlation between specific cardiac parameters (Ventricular Premature Contractions (VPC) number, grade of arrhythmia, heart rate, etc.) detected by electrocardiogram, ultrasound, and Holter monitoring and the development of clinical signs including fainting, sudden death and congestive heart failure in 130 adult Boxers previously diagnosed with heart disease.

Active Grant No: 228: A Comparative Evaluation of the Concealed and Overt Forms of Arrhythmogenic Right Ventricular Cardiomyopathy: Risk Factors Associated with the Development of Symptoms in Dogs with Arrhythmogenic Right Ventricular Cardiomyopathy
Diseases: Heart Disease
Sponsors: American Boxer Charitable Foundation
Researcher:
Kathryn Meurs, DVM, PhD, DACVIM
Breeds: Boxer

Abstract:
The clinical syndrome characterized by ventricular arrhythmias, collapsing episodes, sudden cardiac death and sometimes, heart failure in the Boxer dog was previously referred to as Boxer Cardiomyopathy. More recent studied have demonstrated striking similarities, including the inheritance, to a human disease called Arrhythmogenic Right Ventricular Cardiomyopathy (ARVC). The familial nature as well as the devastating outcome of the disease has lead to significant interest in developing screening methods for asymptomatic dogs prior to use for breeding. Screening is to develop a blood test are underway. However, although these screening methods will detect affected dogs, it appears that not all dogs that are affected will ever develop clinical signs. Unfortunately, these dogs may be removed from showing or breeding programs because of their abnormal status. The objective of this study is to evaluate asymptomatic and syncopal Boxers for findings that may relate to the development and presence of symptoms, including ventricular premature couple number, grade of arrhythmia, left and right ventricular size and function, BNP, Troponin I and family history.

Completed Grant No: 2303: Molecular Analysis of Familial Ventricular Arrhythmias in the Boxer Dog
Disease: Heart Disease
Sponsor: American Boxer Charitable Foundation
Researcher: Kathryn Meurs, DVM, PhD, DACVIM
Breed: Boxer
Funding: ABCF - $47,167 CHF funding - $31,346; Total: $78,613

Abstract:
A heart muscle disease in the Boxer dog was initially documented in the 1980s and referred to as Boxer cardiomyopathy. Studies have confirmed that this is an inherited disease primarily characterized by an electrical disturbance in the heart that may lead to collapsing episodes and sudden death. There is increasing demand for a screening test that could be used to evaluate dogs for the disease before they are selected for breeding purposes. Unfortunately, many of the clinical abnormalities do not become apparent until the dog is several years old. Therefore, there is significant interest in developing a DNA test that could be performed before a dog is selected for breeding. The study proposed here is a continuation of a study in which 268 Boxers were evaluated by physical examination, echocardiography and ambulatory electrocardiography. Dogs have been classified as affected, equivocally affected or unaffected; pedigrees and DNA samples have been collected. Three-generation families have been identified. Linkage and candidate gene analysis will now be performed using canine markers and these DNA samples. The identification of a genetic marker linked to familial ventricular arrhythmias will be the first stem pin the development of a DNA screening test.

Completed Grant No: 2429: The Assessment of Ejection Murmurs in the Boxer Dog
Disease: Heart Disease
Sponsor: American Boxer Charitable Foundation
Researcher: Kathryn Meurs, DVM, PhD, DACVIM
Breed: Boxer
Funding: ABCF - $10,260; CHF matching - $10, 260 Total $20,520