iPS-technology and patient specific disease models

iPS 技术和患者特定疾病模型

• 批准号: 8939854
• 负责人: Manfred Boehm
• 金额: $ 65.17万
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8939854
• 关键词: Alkaline Phosphatase; Cardiac Myocytes; Cardiovascular Diseases; Cell Line; Cells; Characteristics; Communities; Development; Disease; Disease model; Endothelial Cells; Exhibits; Gene Mutation; Genetic; Hematopoietic; Human; In Vitro; Individual; Job' s Syndrome; Mediating; Patients; Peptide Hydrolases; Pharmaceutical Preparations; Protocols documentation; Smooth Muscle Myocytes; Syndrome; System; Vascular Diseases; Vascular Endothelial Cell; Zinc Fingers; base; cell type; homologous recombination; human embryonic stem cell; induced pluripotent stem cell; monocyte; nestin protein; stem cell technology; vector

The development and application of iPS technology promises to revolutionize the understanding and treatment of disease. In particular, it has previously been extremely difficult to obtain sufficient amounts of the relevant human cell types, including cardiac myocytes, endothelial cells and vascular smooth muscle cells, to explore the mechanisms underlying cardiovascular diseases. As such, the ability to reprogram patient specific iPS cells to create cell based disease models represents a major and important paradigm shift for our community. We successfully generated in excess of 30 human iPS cell lines, including approximately 10 patient specific cell lines. To date, these patient specific cell lines have included individuals with Jobs syndrome, as well as an additional rare syndrome characterized by a circulating monocyte deficiency. We have been successful using both lentiviral and retroviral delivery of the four requisite pluripotent factors. We have also extensively characterized many of the human iPS cell lines generated. Our iPS cell lines exhibit characteristics of human embryonic stem cells (ESC) including the ability to form embroid bodies. In addition, these cells express high levels of alkaline phosphatase , Oct4, and Nanog, as well as the human ESC markers SSEA4 and Tra-1-60. Furthermore, we have confirmed that these cells manifest a normal karyotyp and have already begun to develop protocols for differentiation of these iPS cells down various ectodermal, mesodermal and endodermal lineages as assessed by the respective expression of Nestin, Runx1 and GATA4. Successful differentiation of iPS cells towards lineages that are relevant for cardiovascular disease, including endothelial cells, has already been achieved.

iPS技术的发展和应用有望彻底改变对疾病的理解和治疗。特别是，以前极难获得足够量的相关人类细胞类型，包括心肌细胞、内皮细胞和血管平滑肌细胞，以探索心血管疾病的潜在机制。因此，重新编程患者特异性iPS细胞以创建基于细胞的疾病模型的能力代表了我们社区的重大和重要的范式转变。 我们成功地产生了超过30个人iPS细胞系，包括大约10个患者特异性细胞系。迄今为止，这些患者特异性细胞系包括患有乔布斯综合征的个体，以及以循环单核细胞缺乏为特征的额外罕见综合征。我们已经成功地使用慢病毒和逆转录病毒交付的四个必要的多能因子。我们还广泛地表征了所产生的许多人iPS细胞系。我们的iPS细胞系表现出人类胚胎干细胞（ESC）的特征，包括形成胚状体的能力。此外，这些细胞表达高水平的碱性磷酸酶、Oct 4和Nanog，以及人ESC标志物SSEA 4和Tra-1-60。此外，我们已经证实这些细胞表现出正常的核型，并且已经开始开发用于将这些iPS细胞分化为各种外胚层、中胚层和内胚层谱系的方案，如通过巢蛋白、Runx 1和GATA 4的各自表达所评估的。已经实现了iPS细胞向与心血管疾病相关的谱系（包括内皮细胞）的成功分化。