Hematopoiesis from Normal and Patient-Derived Induced Pluripotent Stem Cells

描述（由申请人提供）：该项目涉及NHLBI RC 2 GO申请，标题为“表征通过重编程人类胚胎和诱导多能干细胞衍生的分化心脏、肺和血细胞”。“通过重新编程人类体细胞来产生诱导多能干细胞（iPSC）的新兴技术有望彻底改变生物医学研究和临床医学。通过体外培养方法，iPSC可以分化成来自所有三个胚层的多种细胞类型。这就提出了一种可能性，即患者来源的iPSC可用于为许多人类疾病的研究创造相关组织。此外，iPSC可以提供起始材料来制造用于输血和再生疗法的可移植细胞。然而，这一领域还处于起步阶段，为了实现这些令人兴奋的长期前景，必须解决许多核心问题。该提案旨在促进iPSC在正常和病理性造血研究中的应用。多位在造血、胚胎干细胞/iPSC生物学、染色质生物学、临床血液学、生物信息学、细胞库和生物伦理学/法规事务等领域具有广泛专业知识的研究人员将共同致力于以下全球性问题：1）造血发育潜力在不同正常iPSC克隆之间可能存在差异的机制; 2）iPSC衍生的造血前体在细胞表型、基因表达和表观遗传特征方面与正常造血前体相似的程度; 3）造血疾病表型是否可以通过体外操作患者来源的iPSC来重现。我们将使用新的方法来创建和培养iPSCs，并使用最先进的工具来分析和操纵它们的常驻基因组。对这些问题的追求将作为一个框架，在这个框架中，我们的大型儿科机构的研究人员可以创建，分析，储存和分配来自任何感兴趣患者的iPSC。如果成功，这项工作将有助于加速iPSCs在人类疾病研究和治疗中的实际应用。这项工作将在费城儿童医院进行，并分包给宾夕法尼亚州立大学（州立学院，PA）和Coriell医学研究所（Camden，NJ）。该项目将创造6个新的就业机会，从而刺激美国东北部三个不同地区的经济。 公共卫生相关性：努力更好地了解来自患者的诱导多能干细胞（iPSC）的血液生产将增强我们对血液疾病的理解并产生新的治疗方法。此外，这项工作可以为研究许多正常组织及其相关疾病的起源创造新的一般范式。

DESCRIPTION (provided by applicant): This project addresses the NHLBI RC2 GO application entitled "Characterizing Differentiated Heart, Lung, and Blood Cells Derived by Reprogramming Human Embryonic and Induced Pluripotent Stem Cells." Emerging technologies to generate induced pluripotent stem cells (iPSCs) by reprogramming human somatic cells promises to revolutionize biomedical research and clinical medicine. Through in vitro culture methods, iPSCs can be differentiated into numerous cells types derived from all three germ layers. This raises the possibility that patient-derived iPSCs can be used to create relevant tissues for the study of many human disorders. In addition, iPSCs may provide starting material to manufacture transplantable cells for transfusion and regenerative therapies. However, the field is in its infancy and many core questions must be solved in order to realize these exciting long-term prospects. This proposal seeks to advance the use of iPSCs for the study of normal and pathological hematopoiesis. Multiple investigators with broad areas of expertise in hematopoiesis, embryonic stem cell/iPSC biology, chromatin biology, clinical hematology, bioinformatics, cell banking and bioethics/regulatory affairs will work together to pursue the following global issues 1) Mechanisms by which hematopoietic developmental potential might vary between different normal iPSC clones; 2) The extent to which iPSC-derived hematopoietic precursors resemble normal ones with respect to cellular phenotypes, gene expression and epigenetic signatures; 3) Whether hematopoietic disease phenotypes can be recapitulated by in vitro manipulation of patient-derived iPSCs. We will execute these studies using novel methods to create and culture iPSCs and state-of-the art tools to analyze and manipulate their resident genomes. Pursuit of these problems will serve as a framework in which to develop a facile infrastructure where investigators at our large pediatric institution can create, analyze, bank and distribute iPSCs from any patient of interest. If successful, this work will help to accelerate practical applications of iPSCs for the study and treatment of human diseases. This work will be based at Children's Hospital of Philadelphia with subcontracts to The Pennsylvania State University (State College, PA) and The Coriell Institute for Medical Research (Camden, NJ). The project will create 6 new jobs, thereby stimulating the economy in three different regions of the Northeastern United States. PUBLIC HEALTH RELEVANCE: Efforts to better understand blood production from patient-derived induced pluripotent stem cells (iPSCs) will enhance our understanding of blood disorders and generate new therapeutic approaches. Additionally, this work could create new general paradigms for studying the genesis of many normal tissues and their associated diseases.