Examination of the Effects of Cannabidiol on Canine Neoplastic Cell Apoptosis/Autophagy and Potential for Chemotherapy Resistance or Sensitivity

Currently the use of cannabidiol (CBD) rich extracts for canine oncology patients is common, yet there is no data in canine oncology regarding the effects of CBD on canine cancer cells. Oncologists are wary of CBD use in their patients due to a lack of knowledge regarding the effects of CBD during chemotherapy.

Initial studies on cytotoxicity by the research team show that CBD has cytotoxic activity on a variety of canine cancer cell lines at modest concentrations in the laboratory. These effects cause apoptosis, or programmed cell death, within a very short time frame, suggesting a discrete mechanism.

The objective of this study is two-fold; 1) to determine if co-treatment of cancer cells with a common chemotherapeutic (doxorubicin) and CBD at varying concentrations affects chemotherapeutic cytotoxicity, and 2) to examine the molecular framework of the cell death response looking at the most commonly implicated pathways targeted in canine cancer treatment, including mechanisms of cell signaling and autophagy (removal of damaged cells).

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 02643-A

RESEARCHERS

Joesph J. Wakshlag, DVM, PhD; University of Florida
Amount: $2,500

Discovery of Novel Biomarkers of Canine Atopic Dermatitis through Lipid Profiling

Canine atopic dermatitis (CAD) is a common allergic skin disease of dogs with a strong genetic basis. CAD can severely affect the health and well-being of dogs and current diagnosis of CAD requires time-consuming and expensive procedures for the owner. Furthermore, the molecular mechanisms underlying this condition are not well understood.

Evidence from human studies suggests that several variants of atopic dermatitis (AD) exist with different mechanisms and responses to treatment. Therefore, new approaches to identify molecular markers that can help with better diagnosis and management are warranted. CAD and human AD are associated with changes in the composition of lipids in the epidermis which may precede the inflammation or result from the inflammation.

The investigators will analyze the lipid composition of the epidermis and blood of healthy dogs in comparison to dogs with CAD using a novel analytical method developed by their interdisciplinary team. The results of this work could lead to new, minimally-invasive tests for the diagnosis of CAD and for the prediction and monitoring of the response of CAD patients to treatment.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 02651

RESEARCHERS

Harm HogenEsch, DVM, PhD; Purdue University
Amount: $2,500

Embracing Polygenicity of Common Complex Disease in Dogs: Genome wide Association of Cruciate Ligament Rupture

Cruciate ligament rupture (CR) is a common disabling, degenerative condition of the knee. It places a large financial burden on the American public. Inflammation of the stifle and fraying of cruciate ligament fibers, particularly in the cranial cruciate ligament, eventually leads to ligament rupture with associated stifle instability in affected dogs. CR is a moderately heritable, complex disease with genetic and environmental risk.

CR is common in certain breeds, such as the Labrador Retriever, and rare in other breeds. There is a critical gap in knowledge regarding the genetic contribution to CR, as the number of genes influencing disease risk has never been studied in detail.

Our main goal is to comprehensively analyze the genetic features of the disease across the genome and use this knowledge to develop a genetic test for CR disease risk using genomic prediction. We aim to robustly estimate heritability, analyze the genetic architecture of CR, and advance genetic testing using genomic prediction in the Labrador Retriever, the most common purebred dog breed. The rationale for this work is that detailed knowledge of the genetic features of CR will advance development of a genetic test for CR risk using genomic prediction. This work will fundamentally advance knowledge of the genetic architecture of CR, a very common canine disease. Consequently, such knowledge will provide an invaluable guide to future research into other canine complex diseases. CR genetic testing would enable early identification of at-risk dogs for precision medical care, and selective breeding to reduce the disease burden.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 02664

RESEARCHERS

Peter Muir, BVSc, PhD; University of Wisconsin, Madison
Amount: $2,500

Targeted next generation sequencing panel for comprehensive testing of vector-borne pathogens

Diagnosing vector-borne disease (VBD) in dogs can be difficult for a number of reasons. First, there are many different disease-causing agents that can be transmitted from ticks/fleas, and the clinical signs caused by these agents in dogs can overlap. Additionally, because ticks/fleas can harbor more than one agent at a time, multiple pathogens may be passed to a dog with a single vector bite, resulting in co-infections.

VBD infections can initially present with non-specific signs, such as fever, lethargy, vomiting, diarrhea, and/or respiratory signs. Severe cases can be associated with neurologic signs. These signs can be a diagnostic conundrum. While initial blood work can be helpful and suggest VBD, it does not determine the infecting agent.

This study will develop a comprehensive next generation sequencing panel to detect and identify major VBD agents known to cause disease in dogs and to aid in diagnosis of active infections. Additionally, through parallel sequencing with this method, this panel will incorporate testing for additional infectious diseases that may cause GI, respiratory, or neurologic signs in dogs. The comprehensive nature of this sequencing panel should be a useful tool for surveillance of infectious diseases in the canine population for rapid identification of VBD in dogs and protection of pet owners from such zoonotic diseases.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 02553

RESEARCHERS

Rebecca Wilkes, DVM; Purdue University
Amount: $2,500

Clinical Trial for Evaluation of Propranolol and Doxorubicin in the Treatment of Canine Hemangiosarcoma

Canine hemangiosarcoma is a largely incurable cancer in dogs, and treatment approaches to improve outcomes have remained relatively stagnant over the past few decades. Treatment remains a challenge partly because the cancer is frequently detected at an advanced stage and because these tumors are often resistant to chemotherapies. Recently published reports showed that propranolol, a drug used to treat heart disease in humans and dogs, substantially increased the survival time of human angiosarcoma patients when used in combination with standard of care treatments. Propranolol was also shown to sensitize hemangiosarcoma cells to doxorubicin, providing a more effective way to kill tumor cells. Because angiosarcoma is strikingly similar to canine hemangiosarcoma, this multi-institutional clinical trial has been designed to determine the efficacy of propranolol in dogs with hemangiosarcoma when used in combination with surgery and chemotherapy.

The main goal of the study is to establish whether propranolol in combination with doxorubicin following surgery improves outcomes for dogs when compared to the use of chemotherapy and surgery alone. The investigators will also evaluate the plasma concentrations of propranolol achieved during dosing to assess whether the levels of propranolol correlate to survival times. If successful, the findings from this approach will be rapidly conveyed to the veterinary community, and the guidelines provided to clinicians for the use of propranolol and doxorubicin for the treatment of canine hemangiosarcoma.

Co-investigators: David R. Brown, PhD, University of Minnesota; Michael O. Childress, DVM, MS, Purdue University; Jennifer Mahoney, DVM and Pascale Salah, DVM, University of Pennsylvania

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 02534

RESEARCHERS

Erin Dickerson, PhD and Brian Husbands, DVM; University of Minnesota
Amount: $10,000

Investigation into Diet-Associated Dilated Cardiomyopathy in Dogs

Dilated cardiomyopathy (DCM) is a serious disease of the heart muscle whereby the heart becomes enlarged with weak contractions. DCM can result in  abnormal heart rhythms, congestive heart failure or sudden death. In dogs, DCM most often occurs in large- and giant-breeds, such as Doberman  Pinschers, Boxers, Irish Wolfhounds, and Great Danes; in these dogs, survival time after diagnosis is often only months, even with aggressive medical  therapy.

Recently, veterinary cardiologists have recognized DCM more frequently in all breeds of dogs including mixed breeds, and even those not  usually associated with DCM. There is suspicion that the disease in some dogs is associated with boutique, exotic ingredient, or grain-free (BEG) diets. Some affected dogs on such diets have shown reversal or improvement of their disease after changing their diet, supporting a potential association  between consumption of a BEG diet and development of DCM. A specific cause, however, has not been identified, despite extensive nutritional testing of  the dog foods and the canine patients. Moreover, the extent of the problem is unknown because only dogs that are symptomatic for DCM have been  reported. It is possible that more dogs may be affected but not yet showing signs of heart disease.

To investigate the extent of diet-associated heart  problems in dogs, this multi-institutional team of veterinary cardiologists and nutritionists will prospectively screen a large population of apparently  healthy dogs for DCM and compare important cardiac disease measures, including ultrasound of the heart, blood biomarker and taurine concentrations,  and the frequency of DCM in dogs eating BEG versus non-BEG diets.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 02661

RESEARCHERS

Darcy Adin, DVM; University of California, Davis
Amount: $5,000

Addison’s Disease and Symmetrical Lupoid Onychodystrophy in Bearded Collies Provide Common Ground for Identifying Susceptibility Loci Underlying Canine Autoimmune Disorders

Hypoadrenocorticism or Addison’s disease (AD) is a life-threatening condition that afflicts multiple dog breeds and results from autoimmune destruction of the adrenal glands. Similarly, another canine autoimmune condition that causes pain and suffering is Symmetrical Lupoid Onychodystrophy (SLO).

Both AD and SLO are postulated to be complexly inherited and preliminary data suggest a common set of susceptibility genes working in concert with additional genes that determine expression of either disease. For the study of AD and SLO the investigators will focus on the Bearded Collie breed due to its relatively high prevalence of both conditions and a genomic structure favorable for identifying variations in the DNA.

The investigators will scan the entire canine genome using genetic markers coupled with whole genome sequencing to identify chromosomal regions that harbor genetic changes contributing to disease manifestation. The disease risk conferred by each of these genetic variants, or quantitative trait loci (QTL), will then be calculated to develop a tool for selecting sires and dams early in life, thereby allowing breeders to choose mating pairs that will produce offspring with a low likelihood of developing AD and SLO.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 02488

RESEARCHERS

Anita Oberbauer, PhD; University of California, Davis
Amount: $15,000

Developing a Next Generation Sequencing Diagnostic Platform for Tick-Borne Diseases

Diagnostic tests based on the detection of DNA from harmful organisms in clinical samples have revolutionized veterinary medicine in the last decades. Currently, diagnostic panels for several vector-borne organisms are available through universities and private labs in the USA and abroad. However, the vast majority of results from sick dogs are negative, which frustrates veterinarians and dog owners trying to reach a definitive diagnosis. These panels are based on the detection of previously known DNA sequences of each pathogen, which limits their ability to detect novel organisms.

In this study, the investigators will adapt high-throughput nextgeneration sequencing (NGS) to the detection of tick-borne bacteria in dog blood in an effort to overcome the limitations of current diagnostics for tick-borne diseases. If successful, increasing the capabilities of NGS to detect infected dogs and to accurately determine which bacteria are responsible for disease will support the development of a better diagnostic tool to simultaneously advance canine and human health. This work expands on Dr. Diniz’s previous CHF-funded study #02292.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 02528

RESEARCHERS

Pedro Diniz, DVM, PhD; Western University of Health Sciences
Amount: $5,000

Targeting the Cancer Epigenome: The Effect of Specific Histone Lysine Methyltransferase Inhibition in Canine B-Cell Lymphoma

Canine lymphoma is one of the most common cancers in dogs. While some breeds appear more at risk than others, all can be affected. Although it is often treatable, canine lymphoma can rarely be cured. A continued understanding of the mechanisms causing lymphoma in dogs and identification of novel therapies are needed to improve survival in dogs with lymphoma.

One area of research that has been actively explored and provided exciting breakthroughs for human lymphoma is epigenetics, or alterations in how genes are turned on and off independent of the DNA sequence. One way in which this occurs is due to modifications of the proteins that interact with DNA called histones. Various modifications to these histones can result in genes being turned on or off, leading to the development of cancer. One particular enzyme that modifies histones, EZH2, has been found to play a role in some human lymphomas. However, this has been unexplored in canine lymphoma. Given the striking similarities between human and canine lymphoma, the objective of this work is to characterize the function and role of EZH2 in canine lymphoma.

The investigators will utilize an EZH2 inhibitor to study EZH2 in canine lymphoma cells. The information obtained from this study will help guide the future development of this targeted inhibitor for use as a novel therapy to treat canine lymphoma.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 02309

RESEARCHERS

Angela McCleary-Wheeler, DVM, PhD; University of Missouri, Columbia
Amount: $5,000

Efficacy of Cannabidiol (CBD) for the Treatment of Canine Epilepsy

Epilepsy is the most common neurologic condition in dogs. Approximately 20-30% of dogs receiving standard therapy remain uncontrolled for their seizures. Additionally, the side effects of the antiepileptic drugs (AED) are often unacceptable. Thus, there is a need for an AED that is efficacious with minimal side effects. Cannabidiol (CBD), a prominent non-psychotropic component of the Cannabis sativa plant, has been shown to have anti-convulsant properties. While CBD offers promise as a treatment for canine epilepsy, controlled studies are needed to prove its effectiveness.

In this randomized, double-blinded, placebo-controlled, crossover clinical trial, client-owned dogs with uncontrolled epilepsy will be enrolled following a full seizure evaluation, including bloodwork and magnetic resonance imaging. The canine patients will first receive either a placebo or CBD in addition to their standard AED protocol and then the opposite drug in this crossover designed study. Seizure frequency and medication side effects will be monitored by owners using a seizure log and questionnaire. Regular CBD plasma concentrations, routine bloodwork and serial physical examinations will be monitored by the investigator.

The primary goal of the study is to determine the efficacy of CBD in the treatment of canine epilepsy. If CBD is effective in decreasing seizure frequency, it has the potential to improve the quality and length of life for dogs with uncontrolled epilepsy, and add a much-needed tool for veterinarians in the treatment of canine epilepsy.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 02323

RESEARCHERS

Stephanie McGrath, DVM, MS; Colorado State University
Amount: $10,000