Development of iPS Cells for Treatment of Hemoglobinopathies

描述（由申请人提供）：拟议的POI是一项多机构赠款，将开发用于治疗镰状细胞病（SCD）和地中海贫血（<$地中海贫血）的干细胞疗法。- 塔尔）以及其他血红蛋白病，使用患者来源的体细胞并将它们重编程为诱导多能干（IPS）细胞，所述诱导多能干（IPS）细胞将使它们的突变得到校正并最终分化为造血干细胞（HSC）以重建患者的造血系统。开发用于治疗血红蛋白病（世界范围内最常见的遗传性疾病）的有效细胞疗法将显著改善患有SCD和B-地中海贫血的个体的生活质量，所述SCD和B-地中海贫血在非洲、地中海、中东、非洲和非洲的人民中是常见的。这一提议将检验一种有效的细胞和基因疗法可以实现的假设，在PPG的背景下，通过患者来源的iPS细胞的产生、修饰和造血分化。这将通过以下项目完成：项目1将涉及使用phiC 31整合酶介导的质粒的序列特异性整合将患者的体细胞转化为IPS细胞，所述质粒携带2A肽连接的Oct 4、Sox 2、Klf 4和cMyc重编程cDNA，或通过使用小激活双链RNA（saRNA）瞬时增强这些重编程基因的表达。项目2将涉及在存在或不存在靶向锌指核酸酶（ZFN）或其他大范围核酸酶的情况下，通过使用经典同源重组（HR）或通过基于寡核苷酸/多核苷酸的小片段同源置换（SFHR）的序列特异性修饰来校正体细胞和iPS细胞中的致病突变。项目3将涉及将未校正和校正的iPS细胞暴露于引导造血分化的条件下，以产生具有移植和重建造血系统能力的HSC。在本PPG过程中，所有项目将开发无异源系统以优化安全性。项目中的科学将通过一个行政核心（核心A）和两个科学核心（核心B：细胞和分子生物学，以及核心C：细胞移植和分析）来增强。 公共卫生相关性：该计划项目将侧重于开发治疗镰状细胞性贫血和B型地中海贫血的方法，这是世界上最常见的遗传病。它的目的是通过遗传校正患者细胞来产生用于移植的干细胞，从而避免由于组织不相容性而导致的排斥反应，从而设计治疗这些疾病的新方法。纠正这些疾病将显着提高患者的生活质量，并减少他们对医疗保健系统造成的社会和经济负担。

DESCRIPTION (provided by applicant): The proposed POI is a multi-institutional grant that will develop a stem cell based therapy for the treatment of sickle cell disease (SCD) and ¿-thalassemia (¿-thal) as well as other hemoglobinopathies, using patient derived somatic cells and reprogramming them into induced pluripotent stem (IPS) cells that will have their mutations corrected and ultimately differentiated into hematopoietic stem cells (HSCs) to reconstitute the patient's hematopoietic system. Development of an effective cellular therapy for the treatment of hemoglobinopathies, the most common inherited diseases worldwide, would significantly improve the quality of life of individuals afflicted with SCD and B-thalassemia that are common among the peoples of Africa, the Mediterranean, the Middle East, and Asia as well as their descendents in the U.S. This proposal will test the hypothesis that an effective cellular and genetic therapy for these diseases can be achieved in the context of this PPG through the generation, modification, and the hematopoietic differentiation of patient derived iPS cells. This will be accomplished through the following Projects: Project 1 will involve the conversion of a patient's somatic cells into IPS cells using phiC31 Integrase-mediated, sequence-specific integration of a plasmid carrying 2A peptide linked Oct4, Sox2, Klf4, and cMyc reprogramming cDNAs or by using small activating double stranded RNA (saRNA) to transiently enhance the expression of these reprogramming genes. Project 2 will involve correction of the disease causing mutations in the somatic cells and the iPS cells by sequence specific modification using either classical homologous recombination (HR) or by oligo/polynucleotide-based small fragment homologous replacement (SFHR) in the presence or absence of targeted zinc finger nucleases (ZFNs) or other meganucleases. Project 3 will involve exposure of uncorrected and corrected iPS cells to conditions to direct hematopoietic differentiation to generate HSCs which have the capacity to engraft and reconstitute the hematopoietic system. In the course of this PPG, all Projects will develop xeno-free systems to optimize safety. The science in the Projects will be augmented by an administrative (Core A) and 2 scientific Cores: Core B: Cell and Molecular Biology, and Core C: Cell Transplantation and Analysis. PUBLIC HEALTH RELEVANCE: This Program Project will focus on the development of a therapy for sickle cell anemia and B-thalassemia the most common genetic diseases worldwide. It aims to devise new methods of treating these diseases by genetically correcting patient cells to generate stems cells for transplantation, thus avoiding rejection due to histo-incompatibility. Correcting these diseases would significantly improve the quality of life among afflicted individuals and decrease the social and economic burden that they impose on the healthcare system.