Using human ESCs as a genetic model for PNH and other blood diseases

使用人类 ESC 作为 PNH 和其他血液疾病的遗传模型

• 批准号: 7157979
• 负责人: Guibin Chen
• 金额: $ 5.8万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-01-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7157979
• 关键词: CD34 molecule; NOD mouse; SCID mouse; clinical research; gene mutation; genetic models; genetically modified animals; glycosylphosphatidylinositols; hematopoiesis; hematopoietic stem cells; human embryonic stem cell line; human tissue; membrane proteins; mixed tissue /cell culture; model design /development; molecular pathology; nucleic acid structure; paroxysmal nocturnal hemoglobinuria; postdoctoral investigator; protein deficiency; stromal cells; transfection /expression vector

DESCRIPTION (provided by applicant): Paroxysmal Nocturnal Hemoglobinuria (PNH) is a clonal disorder of hematopoietic stem cells (HSCs) acquiring mutations in the PIG-A gene. Clonal PIG-A mutations are found in nearly all PNH patents tested resulting in the lack of all GPI-anchored proteins (GPI-APs) in affected HSCs and all the derived hematopoietic progeny. Despite evidences that the biochemical and molecular mechanisms for PNH have been brilliantly elucidated, mechanisms of PIG-A mutant clonal dominance in PNH patients and the close relationship of PNH to other marrow failure diseases such as aplastic anemia (AA) and myelodysplasia syndrome (MDS) are still unknown. Monitoring the PIG-A mutations in patients is impossible at early stage before the onset of clinical features. Creating a PIG-A mutation in human HSCs from healthy volunteers is not feasible currently and existing Pig-a null mice lacking GPI-APs in. blood cells have not replicated the PNH symptoms seen in patients. The lone term objective of this research is understanding effects of GPI-APs in bone marrow failure syndrome and hematopoesis. In response to PA-05-013, a human ESC-based [NIH-approved, WA01(H1)], prospective experimental system is proposed here to investigate effects of the PIG-A/GPI-AP deficiency in human hematopoietic cells. The NIH-approved human ESC that contain an induced PIG-A mutation and are GPI-APs deficient will be used, and effects of PIG-A/GPI-AP deficiency using human ESC-initiated hematopoiesis systems recently developed will be examined. The successful completion of this project may also provide a novel genetic model to investigate normal and abnormal human hematopoiesis and HSCs. Improved understanding of PIG-A/GPI-AP deficiency in human hematopoiesis and HSCs will in turn help us to improve and develop new treatments for PNH and other related blood diseases such as AA & MDS.

描述（由申请人提供）：阵发性睡眠性血红蛋白尿症（PNH）是一种造血干细胞（HSC）获得 PIG-A 基因突变的克隆性疾病。几乎所有测试的 PNH 专利中都发现了克隆性 PIG-A 突变，导致受影响的 HSC 和所有衍生的造血后代中缺乏所有 GPI 锚定蛋白 (GPI-AP)。尽管有证据表明 PNH 的生化和分子机制已得到很好的阐明，但 PNH 患者中 PIG-A 突变克隆优势的机制以及 PNH 与其他骨髓衰竭疾病如再生障碍性贫血 (AA) 和骨髓增生异常综合征 (MDS) 的密切关系仍然未知。在临床特征出现之前的早期阶段监测患者的 PIG-A 突变是不可能的。目前，在来自健康志愿者的人类 HSC 中产生 PIG-A 突变是不可行的，并且现有的血细胞中缺乏 GPI-AP 的 Pig-a 缺失小鼠并未复制患者中所见的 PNH 症状。本研究的唯一目标是了解 GPI-AP 在骨髓衰竭综合征和造血中的作用。为了响应 PA-05-013（一种基于人类 ESC 的[NIH 批准的，WA01(H1)]），本文提出了前瞻性实验系统，以研究 PIG-A/GPI-AP 缺陷对人类造血细胞的影响。将使用 NIH 批准的含有诱导 PIG-A 突变且 GPI-AP 缺陷的人类 ESC，并使用最近开发的人类 ESC 启动的造血系统检查 PIG-A/GPI-AP 缺陷的影响。该项目的成功完成也可能为研究正常和异常的人类造血和造血干细胞提供一种新的遗传模型。对人类造血和 HSC 中 PIG-A/GPI-AP 缺陷的进一步了解将有助于我们改进和开发 PNH 和其他相关血液疾病（如 AA 和 MDS）的新疗法。