Globin Gene Expression in Sickle Cell Genotype-Specific iPS cells

镰状细胞基因型特异性 iPS 细胞中的球蛋白基因表达

基本信息

批准号：
8501667
负责人：
DAVID H K CHUI
金额：
$ 220.01万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-05 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8501667
关键词：
Accounting Address Adult Affect African American Animals Blood Blood Cells Cell Lineage Cells Clinical Development Disease Embryo Erythroid Erythroid Cells Erythroid Progenitor Cells Erythropoiesis FDA approved Fetal Development Fetal Hemoglobin Gene Expression Gene Expression Profile Generations Genes Genetic Genetic Variation Genotype Globin Goals Growth Hemoglobin Hereditary Disease Histone Deacetylase Inhibitor Human In Vitro Learning Messenger RNA Modeling Molecular Profiling Morbidity - disease rate Other Genetics Patients Pharmaceutical Preparations Phase Preclinical Drug Evaluation Production Quantitative Trait Loci Rare Diseases Regulation Residual state Sickle Cell Sickle Cell Anemia Sickle Hemoglobin Staging Stem cells System Technology Transgenes Translating Work base cytotoxicity erythroid differentiation fetal genome-wide high throughput screening hydroxyurea induced pluripotent stem cell mortality novel novel therapeutics polymerization pre-clinical response sickle erythroid sickling small molecule stem vector

项目摘要

DESCRIPTION (provided by applicant): Sickle cell anemia, an orphan disease of African Americans, is noted for its extensive morbidity and high mortality. Only one FDA approved drug is available for its pathophysiologically-based treatment. This agent, hydroxyurea, works through its ability to induce fetal hemoglobin (HbF) expression, which thwarts sickle hemoglobin polymerization. Not all patients respond to this treatment, so additional HbF inducing drugs are needed. Our goal is to create the technology for developing high throughput sickle cell anemia-specific induced pluripotent stem cells (iPS) and characterize their directly differentiated progeny. Using a novel excisable reprogramming vector we will generate 'clinical grade' human iPS cells free of any residual reprogramming transgenes. These directly differentiated sickle iPS cells will be used to produce an unlimited supply of erythroid-lineage cells to better understand HbF genetic regulation and perform pre-clinical small molecule drug screens. Specifically, we hypothesize: 1) that 'clinical grade' disease-specific iPS cells can be efficiently and reproducibly derived from peripheral blood cells and induced to normal erythroid differentiation, which includes the expected the spectrum of p-like globin gene expression; 2) major HbF quantitative trait loci impact globin gene expression during embryonic, fetal, and adult erythropoiesis; 3) patients with markedly elevated HbF levels serve as natural models to identify and characterize genetic variations affecting HbF production; 4) high-potency inducers of HbF, either singly or in combination can be studied in iPS-derived erythroid precursor cells. Our aims are: 1) implement an efficient and 'scalable' system for the production of sickle cell anemia-specific iPS cells and use this to recapitulate erythroid-lineage ontogeny in vitro; 2) identify developmental gene expression profile differences between erythroid precursors that produce primarily HbF and those that produce primarily HbA or HbS; 3) determine the effects of the known major HbF quantitative trait loci on globin gene expression in iPS cells; 4) search for novel HbF genetic modifiers associated with HbF levels by examining gene expression in iPS derived erythroid cells; 5) determine which novel therapeutics discovered in high throughput screens can enhance and maintain high level HBG expression in iPS-derived sickle erythroid cells. Ultimately, we hope to translate these findings into clinically efficacious treatments.

描述（由申请人提供）：镰状细胞贫血是非裔美国人的孤儿疾病，以其广泛的发病率和高死亡率而闻名。仅使用一种FDA批准的药物可用于基于病理生理的治疗。该药物羟基脲通过诱导胎儿血红蛋白（HBF）表达的能力来抑制镰状血红蛋白聚合。并非所有患者都对这种治疗做出反应，因此需要额外的HBF诱导药物。我们的目标是创建用于开发高吞吐量镰状细胞贫血特异性诱导多能干细胞（IP）的技术，并表征其直接分化的后代。使用新型的可分解重编程矢量，我们将生成“临床级”人IPS细胞，而没有任何残留的重编程转基因。这些直接分化的镰状IPS细胞将用于产生无限的红细胞系细胞供应，以更好地了解HBF遗传调节并进行临床前小分子药物筛查。具体而言，我们假设：1）“临床级”特异性IPS细胞可以有效地从外周血细胞中得出，并诱导正常的红细胞分化，其中包括预期的Pliel Glopin Glopin基因表达的谱。 2）主要的HBF定量性状基因座影响胚胎，胎儿和成人红细胞生成期间的球蛋白基因表达； 3）HBF水平明显升高的患者是识别和表征影响HBF产生的遗传变异的天然模型； 4）在IPS衍生的红细胞前体细胞中，可以研究HBF的高功率诱导剂。我们的目的是：1）实施一个有效且可扩展的系统，以生产镰状细胞特异性IPS细胞，并使用它来概括体外的红斑和肌。 2）确定主要产生HBF和主要产生HBA或HB的红细胞前体之间的发育基因表达谱差异； 3）确定已知的主要HBF定量性状基因座对IPS细胞中球蛋白基因表达的影响； 4）通过检查IPS衍生的红细胞细胞中的基因表达来搜索与HBF水平相关的新型HBF遗传修饰剂； 5）确定在高吞吐量筛选中发现的哪种新型治疗剂可以增强和维持IPS衍生的镰状细胞细胞中高水平的HBG表达。最终，我们希望将这些发现转化为临床有效的治疗方法。