Improving the Efficiency of Induced Pluripotent Cell (IPS) Generation in the Dog

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Stem cells have recently generated more public and professional interest than almost any subject in biology. These cells have captured the imagination of scientists as a result of the promise offered for a greater understanding of cellular biology and their potential for revealing pathways towards better treatment for chronic illnesses. In veterinary medicine, there is the added advantage of developing a series of reagents that can be used to reduce the number of animals in research. It has recently been demonstrated in humans and mice that adult cells in the body can be 'reprogrammed' to become stem cells by the introduction of four genes or 'factors'. These reprogrammed cells are called Induced Pluripotent Stem Cells (iPS cells) and are of incredible importance because it means that stem cells can be made without the need for deriving them from embryos. Recently, we demonstrated that this can be achieved in the dog using a viral gene delivery approach and we consider that IPS generation will have major beneficial effects, including: 1) We consider that an understanding of stem cell biology will help to understand chronic diseases of the dog. 2) This project will provide the raw materials for us to be able to explore the potential of regenerating diseased tissues. 3) This project will provide the raw materials to be able to test pharmaceutical drugs without the need for experimental animals. 4) For veterinary medicine to advance significantly, these cells and reagents are vital for providing the foundation and building blocks for the next generation of medicines. 5) Ultimately, efficient generation of IPS cells could be applied to generate disease-specific IPS cells that could be applied to better understand the basic mechanisms of disease. However, the current methodology to make iPS cells includes the use of viruses to deliver the factors or genes and is considered to be an inefficient process. For this project we consider that Canine iPS cells can be generated using synthetically modified mRNA delivery (possible combined with small molecule inhibitors) or recombinant proteins instead of viral vectors. Further, this project will analyse the reprogramming events during this process that will help us develop more efficient means of generating canine iPS cells rapidly. The overall aim of this project is to improve the efficiency of iPS generation in the dog, characterize these lines at a molecular level, and explore the mechanisms that maintain pluripotency. We believe that this project will provide the reagents and tools to form the basis of 21st century advances in veterinary medicine. Further, it will provide reagents to help reduce the number of experimental animals used in research.