Modeling chromosome 7 loss in Myelodysplasia-iPSCs

骨髓增生异常-iPSC 中 7 号染色体缺失的建模

• 批准号: 8788296
• 负责人: Eirini Papapetrou
• 金额: $ 41.74万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8788296
• 关键词: Acute leukemia; Affect; Animal Model; Apoptosis; Biological; Biological Assay; Biological Markers; Bone Marrow; Bone Marrow Cells; Candidate Disease Gene; Cell Cycle; Cell Line; Cell Proliferation; Cell physiology; Cell surface; Cells; Cellular biology; Characteristics; Chromosome abnormality; Chromosomes; Chromosomes, Human, Pair 7; DNA Damage; Data; Development; Disease; Dysmyelopoietic Syndromes; Elements; Engineering; Etiology; Gene Targeting; Genes; Genetic; Genetic Engineering; Genomics; Goals; Health; Hematopoiesis; Hematopoietic; Heterogeneity; Human; Human Genome; Incidence; Ineffective Hematopoiesis; Investigation; Lead; Libraries; Mediating; Methodology; Methods; Modeling; Molecular; Molecular Genetics; Molecular and Cellular Biology; Open Reading Frames; Pancytopenia; Pathogenesis; Pathway interactions; Patients; Phenotype; Pluripotent Stem Cells; Preleukemia; Recurrence; Resources; Role; Somatic Cell; Stem Cell Research; Stratification; Technology; Time; Tissue Sample; Undifferentiated; Work; arm; base; bone marrow hyperplasia; cellular engineering; chromosome 7 loss; chromosome 7q loss; clinical decision-making; disease mechanisms study; embryonic stem cell; induced pluripotent stem cell; insight; leukemia; new technology; novel; outcome forecast; palliative; response; reverse genetics; targeted treatment; tool

DESCRIPTION (provided by applicant): Myelodysplastic syndromes (MDS) are clonal hematologic disorders characterized by ineffective hematopoiesis and a propensity for progression to bone marrow (BM) failure or acute leukemia. Despite their relatively high incidence, very little is known about their pathogenesis and their treatment remains mainly palliative. This is largely due to the unavailability of good animal models and the challenges of the ex vivo culture of primary MDS cells. Loss of the entire or part of chromosome 7 [del(7/7q)] is a recurrent cytogenetic abnormality in MDS, associated with unfavorable prognosis, strongly suggesting that one or more critical genes - that so far remain elusive - reside in chromosome 7q. With recent breakthroughs in human pluripotent stem cell (hPSC) research - direct reprogramming and new genetic engineering technologies - reverse genetics in an isogenic setting by precise disruption of genomic elements into their cognate genomic and cellular context - hitherto unthinkable for the human genome - are now a realistic prospect. Our goal is to harness cutting-edge reprogramming and genetic engineering technologies that we and others have developed to establish a novel hPSC-based model to study the cellular, molecular and genetic pathogenesis of myelodysplasia. In preliminary studies, we have derived MDS-iPSC lines with chromosome 7q deletions from patient BM cells and found that they recapitulate potential disease- associated phenotypes: impaired cell proliferation and hematopoietic differentiation. In the proposed study we plan to derive additional del(7q)- as well as isogenic karyotypically normal iPSCs, through reprogramming and chromosome engineering, and characterize their phenotype in the undifferentiated state and following hematopoietic differentiation. To identify genes with a role in MDS pathogenesis, we will perform a screen of candidate genes residing in chromosome 7q for rescue of proliferation in our del(7q)- hPSCs. These studies, using powerful new technologies, will provide a novel valuable resource for the study of myelodysplasia, generate insights into the cellular processes and molecular pathways affected and potentially identify critical genes in the pathogenesis of MDS, bone marrow failure and preleukemia, in general.

描述（由申请人提供）：骨髓增生异常综合征（MDS）是一种克隆性血液疾病，其特征是无效的造血功能和进展为骨髓（BM）衰竭或急性白血病的倾向。尽管其发病率相对较高，但对其发病机制知之甚少，其治疗仍主要是姑息治疗。这主要是由于缺乏良好的动物模型以及原代MDS细胞离体培养的挑战。整个或部分 7 号染色体缺失 [del(7/7q)] 是 MDS 中一种反复出现的细胞遗传学异常，与不良预后相关，强烈表明一个或多个关键基因（迄今为止仍难以捉摸）位于 7q 号染色体上。随着人类多能干细胞 (hPSC) 研究的最新突破 - 直接重编程和新的基因工程技术 - 通过将基因组元件精确破坏到其同源基因组和细胞背景中，在等基因环境中逆转遗传学 - 迄今为止对人类基因组来说是不可想象的 - 现在已成为现实。我们的目标是利用我们和其他人开发的尖端重编程和基因工程技术来建立一种基于 hPSC 的新型模型，以研究骨髓增生异常的细胞、分子和遗传发病机制。在初步研究中，我们从患者 BM 细胞中衍生出具有 7q 染色体缺失的 MDS-iPSC 系，并发现它们重现了潜在的疾病相关表型：细胞增殖和造血分化受损。在拟议的研究中，我们计划通过重编程和染色体工程获得额外的 del(7q) 以及等基因核型正常 iPSC，并表征其未分化状态和造血分化后的表型。为了鉴定在 MDS 发病机制中起作用的基因，我们将筛选位于 7q 染色体上的候选基因，以挽救 del(7q)- hPSC 的增殖。这些研究使用强大的新技术，将为骨髓增生异常的研究提供一种新颖的宝贵资源，深入了解受影响的细胞过程和分子途径，并有可能识别MDS、骨髓衰竭和白血病前期发病机制中的关键基因。