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

最近，干细胞比生物学中的几乎任何学科都引起了更多的公众和专业兴趣。这些细胞激发了科学家的想象力，因为它们有望更好地理解细胞生物学，并揭示更好地治疗慢性病的途径。在兽医领域，还有一个额外的优势，那就是开发一系列试剂，可以用来减少研究中的动物数量。最近在人类和小鼠身上证明，通过引入四种基因或“因子”，人体内的成年细胞可以“重新编程”成为干细胞。这些重新编程的细胞被称为诱导多能干细胞(iPS细胞)，具有难以置信的重要性，因为它意味着干细胞可以在不需要从胚胎中获得它们的情况下制造出来。最近，我们证明了使用病毒基因传递方法可以在狗身上实现这一点，我们认为IPS的产生将产生主要的有益影响，包括：1)我们认为对干细胞生物学的了解将有助于了解狗的慢性病。2)这个项目将为我们提供原材料，使我们能够探索疾病组织再生的潜力。3)该项目将提供能够在不需要实验动物的情况下测试药物的原材料。4)要使兽医取得重大进展，这些细胞和试剂对于为下一代药物提供基础和基石至关重要。5)最终，高效的IPS细胞可以用来产生疾病特异性的IPS细胞，这些细胞可以用来更好地了解疾病的基本机制。然而，目前制造iPS细胞的方法包括使用病毒传递因子或基因，这被认为是一个低效的过程。在这个项目中，我们认为可以用合成修饰的信使核糖核酸(可能与小分子抑制剂结合)或重组蛋白来代替病毒载体来产生犬iPS细胞。此外，本项目将分析这一过程中的重编程事件，这将有助于我们开发更有效的方法来快速生成犬iPS细胞。该项目的总体目标是提高犬iPS的生成效率，在分子水平上表征这些品系，并探索维持多能性的机制。我们相信，该项目将提供试剂和工具，形成21世纪兽医进步的基础。此外，它还将提供试剂，帮助减少用于研究的实验动物的数量。

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.