Induced Pluripotent Cells for Blood Diseases

描述(由申请人提供)：对罕见血液疾病患者的评估揭示了生物学、病理生理学和治疗的基本方面。最近分离出了几种罕见遗传病的受影响基因，包括钻石黑扇贫血、什瓦赫曼钻石综合征和范科尼贫血。事实证明，这些疾病的动物模型的推导很困难，因为小鼠的突变表型在许多情况下确实与人类疾病相似。在这里，我们建议从遗传性血液疾病(包括上面列出的疾病和唐氏综合症)患者中建立人iPS诱导的多能细胞系。我们计划从患者身上获得成纤维细胞，并引入Oct4、Sox2、Klf4和c-myc作为逆转录病毒或慢病毒。IPS系将根据细胞形态和多能标记的表达情况而衍生。这些iPS细胞系将以其形成畸胎瘤的能力为特征。此外，我们计划通过在集落试验中将iPS细胞分化为造血细胞类型，以及通过将分化后的细胞移植到免疫缺陷小鼠体内，来表征疾病的体外生物学特征。将建立一个数据库，以帮助跟踪细胞系，提供有关其特征的信息，并帮助向社区分发。然后，我们计划开发一种筛选系统，以寻找在体外拯救疾病表型的小分子或shRNA。这些化学物质或基因将有助于了解疾病的生物学，确定发病机制，并最终找到新的治疗方法。此外，这项工作将为iPS细胞用于其他器官系统的研究铺平道路。

DESCRIPTION (provided by applicant): The evaluation of patients with rare blood disorders has uncovered fundamental aspects of biology, pathophysiology and therapies. The affected gene for several rare genetic diseases has been recently isolated, including Diamond Blackfan anemia, Shwachman Diamond syndrome, Fanconi's anemia. The derivation of animal models for these diseases has proven difficult, as the mouse mutant phenotype does approximate the human disease in many cases. Here we propose to develop human iPS induced pluripotent cell lines from patients with genetic blood diseases (including the disorders listed above and Down's Syndrome). We plan to obtain fibroblasts from patients with the diseases, and introduce Oct4, Sox2, Klf4 and c-myc as retroviruses or lentiviruses. iPS lines will be derived based on cell morphology and pluripotency marker expression. These iPS cell lines will be characterized for their ability for form teratomas. Furthermore, we plan to characterize the biology of the disease in vitro by differentiating the iPS cells to form hematopoietic cell types in colony assays as well as by transplanting the differentiated cells into immunodeficient mice. A database will be constructed to help track the cell lines, provide information on their characterization, and to aid distribution to the community. We then plan to develop a screening system to find small molecules or shRNAs that will rescue the disease phenotype in vitro. These chemicals or genes will help understand the biology of the disease, establishing the pathogenesis and ultimately finding new therapies. Furthermore, this work should pave the way for the use of iPS cells for the study of other organ systems.