Performing Department
School Of Veterinary Medicine
Non Technical Summary
The long term goal of the Laminitis Basic Research Program at New Bolton Center is to more fully understand the cause and biology of equine laminitis and thereby improve the prevention, diagnosis and treatment of this common and debilitating condition. Laminitis is a devastating disease of hooved animals that results in the destruction of the attachment between the hoof and underlying bone. This results in biomechanical failure of the foot support system and severe pain, sometimes necessitating the humane destruction of the horse. Research on equine disease is hampered by limited funds and the difficulties involved in conducting research in horses, such as ethical concerns of research in companion animals, financial constraints, small numbers of researchers in the field and limited research publications, and the lack of reagents and background data needed to study a species not traditionally used for research. Collaborative efforts to conquer equine laminitis are dependent upon a well-managed and extensive laminitis tissue, sample, and antibody database. The objective of this project is to generate and maintain a Laminitis Discovery Database at New Bolton Center. Serum, laminar tissue, isolated cells that can be used for tissue culture, extracted DNA, RNA and protein, and fixed/embedded tissue that is stored both at New Bolton Center and at external collaborators' facilities will be inventoried and linked to clinical data and research results. This will aid collaborative efforts and will avoid duplication of research. Samples from clinical cases as well as experimental animals will be banked. Proper management of these samples will assure that the information gained from each horse is maximal. The antibody database will facilitate collaborative research by providing information on the use and efficacy of antibodies on equine tissue. Very few commercially-available antibodies, typically generated against human or mouse proteins, have been tested on equine tissues. Antibodies are essential tools for biomedical research and an antibody database for use on equine tissue would enormously benefit researchers of laminitis and other equine diseases. The Laminitis Laboratory is studying the basic cell biology of cells that compose the lamellae of the hoof and that are directly affected by laminitis with the goal of developing a method for the in vitro culture of these cells based upon methods developed for other epithelial cells (such as human skin). Our Laminitis Discovery Database will also bank samples of cells from normal and laminitic horses for the generation of these tissue cultures. This culture system will be used for in vitro studies of laminitis cause and biology and to generate frozen stocks of cells that can be shared with other laminitis researchers. Currently, equine laminitis research relies heavily upon studies that utilize live animals. By developing an in vitro model system for equine laminitis, I hope to reduce the use of horses for terminal research studies and provide a means of focusing research on the cells that are most directly affected by this disease.
Animal Health Component
(N/A)
Research Effort Categories
Basic
(N/A)
Applied
(N/A)
Developmental
(N/A)
Goals / Objectives
The long term goal of the Laminitis Basic Research Program at New Bolton Center is to more fully understand the pathophysiology of equine laminitis and thereby improve the prevention, diagnosis and treatment of this common and debilitating condition. Laminitis results in the destruction of the lamellar basement membrane and dermal-epidermal attachment between the hoof and underlying pedal bone with subsequent biomechanical failure of digital support, sometimes necessitating the humane destruction of the horse. At the first annual meeting of the External Scientific Advisory Board at New Bolton Center in May, 2008, the consensus of all involved was that collaborative efforts to conquer laminitis are dependent upon a well-managed and extensive laminitis tissue, sample, and antibody database. The objective of this project is to establish and maintain a Laminitis Discovery Database. Serum, laminar tissue, cells isolated from laminar tissue, extracted DNA, RNA and protein, and fixed/embedded tissue that is stored both at New Bolton Center and at external collaborators' facilities will be inventoried and linked to clinical data and research results. This will aid collaborative efforts and will avoid duplication of research. Samples from clinical cases as well as experimental animals will be banked. A collaborator, Christopher Pollitt, will store samples from in vivo models of laminitis (oligofructose and hyperinsulinemia models). Tissue from clinical cases of laminitis are needed to correlate with discoveries made using tissue from in vivo and in vitro models of equine laminitis. Proper management of these samples will assure that the information gained from each horse is maximal. The antibody database will facilitate collaborative research by providing information on the use and efficacy of antibodies on equine tissue. Very few commercially-available antibodies, typically generated against human or mouse antigens, have been tested on equine tissues. Antibodies are essential tools for biomedical research and an antibody database for use on equine tissue would enormously benefit researchers of laminitis and other equine diseases.
Project Methods
Database samples are derived from in vivo induction models for laminitis performed by Dr. Chris Pollitt at the University of Queensland, from University-owned horses that are euthanized for other research projects and teaching, and from clinical cases presented for necropsy at New Bolton Center. These samples include serum, plasma, laminar tissue, and cells isolated from laminar tissue. Tissue is either snap frozen in liquid nitrogen to allow protein or RNA extraction at a later time, fixed and embedded in paraffin, or fixed, serially dehydrated with sucrose, embedded in OCT and frozen for cryosectioning. A board-certified veterinary pathologist examines a specimen from each equine foot from which tissue is banked in order to determine the presence and degree of histopathology. The veterinary school is in the process of acquiring state-of-the-art software for sample management (eSample) from Velos, Inc. This system includes storage and inventory management, bar coding and label printing, multi-site specimen network organization, and can link into electronic medical record systems. The grant will be used to contribute to the information technology costs of adopting and maintaining this system, a portion of technician salary and benefits, and supplies for tissue processing and storage such as tubes, an ultra-low freezer, liquid nitrogen, fixatives, and embedding supplies. The Laminitis Laboratory is studying the basic cell biology of equine epidermal lamellar epithelial cells of the hoof and their underlying basement membrane with the goal of developing a method for the in vitro culture of these cells based upon methods developed for other stratified epithelia (such as human skin). This culture system will be used for in vitro studies of laminitis pathophysiology and to generate frozen stocks of basal epithelial cells and feeder cells that can be shared with other laminitis researchers. Currently, equine laminitis research relies heavily upon in vivo model systems. By developing an in vitro model system for equine laminitis, I hope to reduce the use of horses for terminal research studies and provide a means of focusing research on the cells that are most directly affected by this disease. Tissue culture studies are being conducted in collaboration with an epithelial stem cell biologist, Makoto Senoo.