Modeling and correcting human SCID using patient-derived iPS cells

使用患者来源的 iPS 细胞建模和纠正人类 SCID

基本信息

批准号：
8686738
负责人：
Luigi Daniele Notarangelo
金额：
$ 56.95万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-01 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8686738
关键词：
Affect Animal Model Biopsy Cell Line Cells Coculture Techniques Data Defect Development Disease Disease model Fibroblasts Gene Transfer Generations Genes Genetic Hematopoietic stem cells Hereditary Disease Human Immune Immune system Immunodeficient Mouse Immunologic Deficiency Syndromes In Vitro Infection Injection of therapeutic agent Insertional Mutagenesis Investigation Knowledge Lentivirus Vector Life Ligands Lymphocyte Lymphoid Mediating Modeling Mus Mutation Patients Phenotype Reporting Research Personnel SCID Mice Safety Site Skin Source Staging Stem cells Stromal Cells Subfamily lentivirinae System T cell differentiation T-Cell Development T-Lymphocyte Testing cellular engineering design enzyme replacement therapy gene correction gene therapy hematopoietic cell transplantation human disease improved in vivo induced pluripotent stem cell insight lentiviral-mediated leukemia novel novel strategies pluripotency pre-clinical reconstitution repository research study stem cell biology stemness tool

项目摘要

DESCRIPTION (provided by applicant): Severe combined immune deficiency (SCID) includes a variety of genetic defects characterized by severe numerical and functional defects of T lymphocytes, variably associated with defects of B and NK lymphocytes. Differences exist between mice and humans with mutations in orthologue SCID- associated genes. Moreover, for some forms of SCID no animal models are currently available. Finally, SCID have offered proof of principle that gene therapy can cure human diseases; however, leukemic proliferation due to insertional mutagenesis has been reported in 5 SCID patients treated by gene therapy. Altogether, these notions emphasize the importance of studying human models to gain insights into pathological human T cell development and to develop novel and safer approaches to gene therapy. Co-culture of human hematopoietic stem cells (HSCs) with mouse OP9 stromal cells engineered to express the Notch1 ligands Delta-like 1 (Dll1) or Delta-like 4 (Dll4) has been used to study T cell differentiation. Moreover, severely immunodeficient mice can serve as an in vivo system to study T cell development from human HSCs. However, these approaches can be hardly applied to the study of human SCID, given the rarity of this condition that limits access to freshly isolated HSCs from the patients. Induced pluripotent stem cells (iPSCs) represent a novel tool for disease modeling and correction and may be used to overcome technical and feasibility barriers that have so far hampered the study of T cell development in human SCID. We have assembled a team of investigators with specific and complementary expertise in immunodeficiency, iPSC biology, T cell development and integration site analysis upon gene transfer. We have created a large repository of fibroblast cell lines from patients with various forms of SCID. We now intend to generate iPSCs from these patients. We will characterize the stemness and pluripotency profile and verify genetic integrity of patient-derived iPSCs. To model SCID, we will differentiate iPSCs into T lymphocytes both in vitro and in vivo. We also intend to use patient-derived iPSCs as a platform to explore the efficacy and safety of self-inactivating lentiviral vectors in the correction of various forms of SCID both in vitro and in viv in immunodeficient mice. We anticipate that SCID patients-derived iPSCs will represent a novel and robust platform to study human T cell development and for preclinical assessment of efficacy and safety of gene therapy for these diseases.

描述（由申请人提供）：严重联合免疫缺陷（SCID）包括多种遗传缺陷，其特征是 T 淋巴细胞的严重数量和功能缺陷，与 B 和 NK 淋巴细胞的缺陷不同程度地相关。小鼠和人类之间存在直系同源 SCID 相关基因突变的差异。此外，对于某些形式的 SCID 目前还没有可用的动物模型。最后，SCID 提供了基因疗法可以治愈人类疾病的原理证明；然而，在 5 名接受基因治疗的 SCID 患者中报告了由于插入突变导致的白血病增殖。总而言之，这些概念强调了研究人类模型以深入了解人类 T 细胞病理发育并开发新颖且更安全的基因治疗方法的重要性。人类造血干细胞 (HSC) 与经过工程改造以表达 Notch1 配体 Delta-like 1 (Dll1) 或 Delta-like 4 (Dll4) 的小鼠 OP9 基质细胞共培养已用于研究 T 细胞分化。此外，严重免疫缺陷的小鼠可以作为体内系统来研究人类 HSC 的 T 细胞发育。然而，这些方法很难应用于人类 SCID 的研究，因为这种情况很少见，限制了对人类 SCID 的研究。从患者体内新鲜分离出 HSC。诱导多能干细胞 (iPSC) 代表了一种用于疾病建模和校正的新工具，可用于克服迄今为止阻碍人类 SCID 中 T 细胞发育研究的技术和可行性障碍。我们组建了一支研究人员团队，他们在免疫缺陷、iPSC 生物学、T 细胞发育和基因转移整合位点分析方面拥有特定和互补的专业知识。我们创建了一个来自各种 SCID 患者的成纤维细胞系的大型数据库。我们现在打算从这些患者身上产生 iPSC。我们将表征干性和多能性特征，并验证患者来源的 iPSC 的遗传完整性。为了模拟 SCID，我们将在体外和体内将 iPSC 分化为 T 淋巴细胞。我们还打算使用患者来源的 iPSC 作为平台，探索自失活慢病毒载体在体外和体内纠正免疫缺陷小鼠各种形式 SCID 的功效和安全性。我们预计 SCID 患者衍生的 iPSC 将成为研究人类 T 细胞发育以及对这些疾病基因治疗的有效性和安全性进行临床前评估的新颖而强大的平台。