Targeted Next Generation Sequencing Panel for Comprehensive Testing for Reproductive and Neurologic Pathogens of Dogs

The large number of pathogens that contribute to infectious reproductive or neurologic disease and overlap of clinical signs makes using tests for multi-pathogen detection useful. Routinely, several individual or multiplex PCR assays are performed to identify active infection with these agents.

Determining which tests to use for any given case can be difficult. Therefore, investigators propose to validate a comprehensive targeted next generation sequencing (NGS) panel for detection of all known neurologic and reproductive pathogens that infect dogs, as well as other infectious pathogens of dogs, including vector-borne agents previously validated through our work funded by AKC CHF. This takes the guesswork out of proper test selection for infectious disease diagnostics.

The amount of data that can be generated with NGS allows for comprehensive testing, and use of a targeted method (adding primers to the protocol for amplification of targets of interest) reduces the cost to the point that this testing can be effectively used in a veterinary diagnostic lab. More and more veterinary laboratories are purchasing sequencing equipment, so this method has merit to be expanded to additional labs. With the comprehensive nature of the panel, epidemiological evaluation of infectious diseases in the dog population can also be improved.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 03137

RESEARCHERS

Rebecca Wilkes, DVM, PhD; Purdue University
Amount: $3,000

Characterization of hookworm resistance in dogs with a novel diagnostic test for early intervention

The dog hookworm is the most common worm parasite of dogs in the United States. It can cause disease, resulting in suffering and even death in both young and old dogs of all breeds. Dog hookworms can also infect humans who come into contact with hookworm larvae present in dog stool contaminated areas. Hookworm infections in people will cause cutaneous larval migrans, a painful skin condition. Dogs must be dewormed monthly to prevent the ill effects of acquiring hookworms. While these medications were very effective 20-30 years ago, recently they have stopped being fully effective in many cases. This is because hookworms have become resistant.

To know if a dog has resistant hookworms, veterinarians have to use tests performed at a diagnostic lab. The test that is currently used takes 14 days to complete. In this project, we will create a new diagnostic test that can be performed in less than a day. This new test will let the veterinarian know if the medication that they are using against the hookworms will work or not. This project will add to our knowledge about how much resistance we should expect to see in hookworms from dogs that live in the U.S. Knowing how much resistance to expect in a geographical area and knowing how many hookworms within a dog patient are specifically resistant will help veterinarians make important treatment decisions quickly.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 03169

RESEARCHERS

Jeba Jesudoss Chelladurai, BVSc, PhD
Kansas State University
Amount: $3,000

Identifying Cellular Mechanisms of Inflammation During Canine Tick-Borne Diseases

Tick-borne diseases are found in all 50 states of the United States and are the most common vector-borne disease diagnosed in people in the US. The predominant disease is Lyme disease, caused by Borrelia burgdorferi and related species (sensu lato). Other important canine tick-borne diseases include those caused by Anaplasma platys, Anaplasma phagocytophilum (Anaplasmosis), Babesia canis, Babesia conradea and Babesia gibsonii (Babesiosis), and Ehrlichia canis, Ehrlichia chaffiensis and Ehrlichia ewingii (Ehrlichiosis).

Many of these diseases also affect people. Dogs can serve as sentinel species for human disease and there are many areas where the immune responses and disease outcomes are very similar in people and dogs, meaning that important lessons can be learned by sharing information between human and animal health (One Health).

The researchers will further investigate the dog’s immune system to determine which immune cells are responsible for the cure or creation of canine tick-borne disease. Through understanding which cells are responsible for causing disease, the goal is to then specifically target the molecules they produce using immunotherapy or immune modulation to improve treatment of tick-borne diseases in all dogs.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 02383

RESEARCHERS

Christine A Petersen DVM PhD
University of Iowa
Amount: $5,000

Harnessing a Dog’s Own Immune System to Kill Lymphoma Tumor Cells

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.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 01418

RESEARCHERS

Heather M Wilson-Robles DVM
Texas A&M Research Foundation
Amount: $2,000

Defining the Unique Genetic Markers in Dogs That Define Immune Function, Disease Resistance and Tissue Transplantation

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.

Co-sponsored with the AKC Canine Health Foundation, Grant Number: 01771

RESEARCHERS

Bevery Torok-Storb, PhD, DVM
Fred Hutchinson Cancer Research Center
Amount: $2,000