Patient-specific iPS cells to study myeloproliferative disease.

用于研究骨髓增殖性疾病的患者特异性 iPS 细胞。

• 批准号: 7813728
• 负责人: RUDOLF JAENISCH
• 金额: $ 97.94万
• 依托单位: WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-27 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7813728
• 关键词: Abnormal Myeloid Cell; Address; Applications Grants; Area; Biology; Biomedical Research; Biopsy; Cells; Communities; Complex; Derivation procedure; Development; Disease; Engraftment; Genetic; Genetic Predisposition to Disease; Growth; Hematopoietic; Hematopoietic Neoplasms; Hematopoietic System; Hematopoietic stem cells; Hereditary Disease; Human; Human Genetics; In Vitro; Investigation; JAK2 gene; Methods; Mutate; Mutation; Myelogenous; Myeloid Cells; Myeloproliferative disease; NOD/SCID mouse; Oncogenes; Oncogenic; Parkinson Disease; Pathogenesis; Pathology; Patients; Predisposing Factor; Protocols documentation; Signal Pathway; Skin; Somatic Cell; Stem cells; System; Technology; Time; Transplantation; human disease; improved; in vivo Model; induced pluripotent stem cell; mouse model; neoplastic; peripheral blood; preconditioning; public health relevance; research study; stem; success; vector

DESCRIPTION (provided by applicant): The recent success in reprogramming somatic cells into induced Pluripotent Stem (iPS) cells by defined factors has opened exciting possibilities not only for the investigation of complex human diseases in the Petri dish but also for the ultimate application in transplantation therapy. Direct reprogramming provides for the first time the opportunity to generate in vitro and in vivo models of complex disorders such as familial and sporadic Parkinson's disease using patient-specific cells. The experiments proposed in this application are aimed at using the iPS technology to set up an experimental system to study a human genetic disease of the hematopoietic system. The proposal has three aims: 1. Derivation of iPS cells from peripheral blood: We will establish protocols to isolate iPS cells from myeloid cells of human peripheral blood using vectors that can be deleted by transient Cre expression. 2. Characterization of iPS cell-derived hematopoietic cells (HSCs): We will establish robust protocols allowing in vitro differentiation of iPS cell-derived HSCs and assess their potential to engraft in NOD/SCID mice. To improve engraftment efficiency we will precondition the cells by transient expression of reprogramming factors or oncogenes. 3. Biology and genetics if myeloproliferative neoplasm (MPN): We will establish an experimental paradigm to study the biology and genetic predisposition of myeloproliferative neoplastic disorder (MPN), a disorder characterized by a specific JAK2 mutation. The methods developed in aims 1 and 2 will be used to investigate the pathology of MPN such as assessing whether the JAK2 mutation provides a selective advantage to the growth of hematopoietic cells and whether hematopoietic stem cells derived from skin biopsies of the same patient acquire the mutation when differentiated in vitro or when transplanted into NOD/SCID mice.

描述（由申请人提供）：最近通过定义的因素将重编程为诱导的多能茎（IPS）细胞的成功成功，不仅为研究培养皿中复杂人类疾病的研究开辟了令人兴奋的可能性，而且对于在移植治疗中的最终应用。直接重编程为首次提供了使用患者特异性细胞等复杂疾病（例如家族性和零星帕金森氏病）的体外和体内模型的机会。本应用程序中提出的实验旨在使用IPS技术建立实验系统来研究造血系统的人类遗传疾病。该提案有三个目标： 1。iPS细胞从外周血液中推导：我们将使用可以通过瞬态CRE表达来删除的载体将IPS细胞从人外周血的髓样细胞中分离出来的方案。 2。表征IPS细胞衍生的造血细胞（HSC）：我们将建立允许体外分化IPS细胞衍生的HSC的强大方案 点头/SCID小鼠。为了提高植入效率，我们将通过瞬时表达重编程因子或致癌基因来预处细胞。 3。生物学和遗传学如果骨髓增生性肿瘤（MPN）：我们将建立一个实验范式来研究骨髓增生性肿瘤的生物学和遗传易感性 （MPN），一种以特定JAK2突变为特征的疾病。 AIM 1和2中开发的方法将用于研究MPN的病理学，例如评估JAK2突变是否为造血细胞的生长提供了选择性优势，以及在分化在体外或移植到NOD/Scid/Scid/Scid小鼠中时，造血性干细胞是否衍生自同一患者的皮肤活检获得了突变。