Human Pluripotent Stem Cell and Progenitor Models of Cardiac and Blood Diseases

描述（由申请人提供）： 我们的团队在开发用于产生多能干细胞的改进技术，指导其分化为相关组织，以及在NHLBI的两个主要系统（心血管系统和血液）中的疾病建模方面有共同的兴趣。虽然致病性遗传病变已被确定为许多条件，某些先天性和后天性血液疾病继续造成显着的发病率和死亡率。动物和体外模型的局限性与造血系统特别相关，在造血系统中，将基因缺陷工程化到小鼠品系中未能表现出范可尼贫血和唐氏综合征等疾病的主要特征。人类模型将为研究这些疾病和开发治疗方法提供相关系统。我们开创了体细胞重编程的方法，以产生小鼠和人类诱导多能干细胞（IPS），并为胚胎干细胞（ES）/ IPS细胞定向分化为造血谱系带来了相当多的经验。我们希望利用这些新的“人源化”研究工具来补充我们在斑马鱼和小鼠模型中的传统专业知识，以研究造血发育和疾病病理生理学。在这项提案中，我们计划创建和研究人类IPS细胞的遗传性血液疾病，破坏基因组的稳定性（范科尼贫血和先天性角化不良），指定异常核仁或核糖体蛋白（Shwachman-Bodian-Diamond综合征和Diamond-Blackfan贫血），并代表一个宪法'三体与突出的血液和心脏异常（唐氏综合征）。有了这些IPS细胞，我们将探索疾病表型，寻求基因修复策略，并寻找可能改善这些疾病的新疗法。该提案是与马萨诸塞州总医院的心血管研究人员Ken Chien和Kit帕克博士以及哈佛大学的干细胞研究人员Doug Melton合作的R 03申请的一部分，并有三个具体目标： 目的#1：从患有造血系统遗传性和获得性疾病的患者中产生人类诱导多能干细胞。 目的#2：探索疾病特异性IPS细胞的造血表型。 目标3：研究基因修复的方法，并进行化学和遗传筛选，以确定新的小分子和遗传途径，以改善体外疾病表型。

DESCRIPTION (provided by applicant): Our groups have common interests in the development of improved techniques for generating pluripotent stem cells, directing their differentiation into relevant tissues, and in disease modeling in two major systems of central interest to the NHLBI-the cardiovascular system and the blood. While the causative genetic lesion has been identified for many conditions, certain inborn and acquired hematologic disorders continue to cause significant morbidity and mortality. The limitations of animal and in vitro models is particularly relevant to the hematopoietic system, where engineering gene defects into mouse strains has failed to phenocopy cardinal features of diseases like Fanconi anemia and Down Syndrome. Human models would offer a relevant system to study these diseases and to develop therapeutics. We have pioneered methods for somatic cell reprogramming to generate mouse and human induced pluripotent stem cells (IPS) and bring considerable experience to the directed differentiation of embryonic stem (ES)/ IPS cells into hematopoietic lineages. We wish to exploit these new "humanized" research tools to complement our traditional expertise in zebrafish and murine models to study hematopoietic development and disease pathophysiology. In this proposal we plan to create and study human IPS cells for genetic blood diseases that: disrupt genomic stability (Fanconi's anemia and Dyskeratosis congenita), specify aberrant nucleolar or ribosomal proteins (Shwachman-Bodian-Diamond Syndrome and Diamond-Blackfan Anemia), and represent a constitutional' trisomy with prominent hematologic and cardiac anomalies (Down Syndrome). With these IPS cells, we will explore disease phenotypes, pursue strategies for gene repair, and search for novel therapeutics that might ameliorate these conditions. This proposal is part of a collaborative R03 application with Drs Ken Chien and Kit Parker, cardiovascular researchers at the Massachusetts General Hospital, and Doug Melton, a stem cell researcher at Harvard University, and has three specific aims: Aim #1: Generate human induced pluripotent stem cells from patients with genetic and acquired disorders of the hematopoietic system. Aim #2: Explore the hematopoietic phenotypes of disease-specific IPS cells. Aim #3: Investigate methods for gene repair, and pursue chemical and genetic screening to identify novel small molecules and genetic pathways to ameliorate the disease phenotypes in vitro.