Development of normal and dystrophic canine iPS lines

大量的遗传疾病发生在狗身上，主要是由于创始人效应和几百年来近亲繁殖造成的种群瓶颈。这些疾病包括血管性血友病、严重联合免疫缺陷症、x连锁杜氏型肌肉萎缩症、丙酮酸激酶缺乏症、肌型磷酸果糖激酶缺乏症、血友病和嗜睡症。超过58%的狗遗传疾病，如杜氏型肌肉萎缩症，是由同一基因突变引起的人类疾病的真正同源物。此外，在400多个现代犬品种中，许多犬显示出更复杂的多基因疾病的高患病率，包括过敏、癌症、失明、心脏病、白内障、癫痫、髋关节发育不良和耳聋。由于寿命和体型等明显的参数，狗代表了人类疾病的重要模型。在狗身上，生理、疾病表现和临床反应往往比相应的小鼠模型更接近人类。因此，他们提供了一个独特的临床前模型来测试再生医学和组织工程方法对治疗和治愈人类疾病的有用性。这些新方法的关键将是胚胎干细胞（ES）的分化衍生物的使用。最近的研究表明，通过相对简单的重编程步骤，可以从皮肤成纤维细胞中产生自体多能干细胞（iPS），这是一项重大的科学进步，将在基础研究和应用临床医学的许多领域产生深远的影响。我们的中心假设是，犬iPS细胞系的产生和鉴定将加速人类替代组织和器官的发育。为了解决这一假设，提出了以下三个具体目标：(1)从正常和营养不良犬（GRMD）中产生诱导多能干细胞系（iPS）。(2)将对iPS系的基因表达谱进行表征，并与囊胚衍生犬进行比较。(3)评估犬胚胎干细胞和iPS系的体内和体外多能性。

A large number of genetic diseases occur in dogs mainly due to founder effects and population bottlenecks resulting from centuries-old inbreeding practices. These include von Willebrand disease, severe combined immunodeficiency disorder, X-linked Duchenne type muscular dystrophy, pyruvate kinase deficiency, muscle type phosphofructokinase deficiency, hemophilia and narcolepsy. More than fifty-eight percent of dog genetic diseases, such as Duchenne-type muscular dystrophy, are true homologues of human diseases caused by mutations in the same gene. In addition, many of the more than 400 modern dog breeds show a high prevalence of more complex multigenic diseases, including allergies, cancers, blindness, heart disease, cataracts, epilepsy, hip dysplasia and deafness. Because of obvious parameters such as lifespan and size, dogs represent an important model for human disease. In dogs, the physiology, disease presentation and clinical response often parallels that of the human more closely than the corresponding mouse model. They therefore provide a unique preclinical model to test the usefulness of regenerative medicine and tissue engineering approaches to the treatment and cure of Human disease. Pivotal to these new approaches will be the use of the differentiated derivatives of embryonic stem cells (ES). The recent demonstration that is possible to generate autologous, pluripotent stem cells (iPS) from skin fibroblasts, by relatively simple reprogramming steps is a major scientific advance which will have profound ramifications in many areas of basic research and applied clinical medicine. It is our central hypothesis that the generation and characterization f canine iPS cell lines will hasten the development of replacement tissues and organs in human. In order to address this hypothesis the following three specific aims are proposed (1) To generate induced pluripotent stem cell lines (iPS) from normal and dystrophic (GRMD) dogs. (2) Gene expression profiles of iPS lines will be characterized and compared to a blastocyst derived canine (3) To assess the pluripotency of canine ES and iPS lines in vivo and in vitro.