Gene Replacement Therapy in Induced Pluripotent Stem (iPS) Cells for Treatment of

描述（由申请人提供）：UAB干细胞研究所，UAB生物基质工程和再生医学（BERM）中心和UAB造血干细胞处理设施将在一个项目中合作开发利用诱导多能干细胞（iPS）治疗镰状细胞病的基因替代疗法。该方案由两个项目和两个核心组成。在项目1（用于镰状细胞基因替代的人类iPS细胞）中，Townes博士建议将最近在人源化镰状小鼠模型中纠正镰状细胞病的成功转化为人类细胞。该项目的具体目标是：(1)从镰状细胞病患者的皮肤活检样本中产生人类iPS细胞(2)纠正来自患者的iPS细胞中的镰状突变(3)并将纠正后的iPS细胞分化为可移植的造血干细胞，从而产生正常的红细胞。在项目2（犬丙酮酸激酶基因替代的iPS细胞）中，Clint Lothrop博士提出通过犬iPS细胞的基因替代来纠正遗传性贫血（犬丙酮酸激酶缺乏症）的大型动物模型。该项目的具体目的是：(1)从遗传性丙酮酸激酶（PK）缺乏症犬的皮肤活检样本中制备犬的iPS细胞(2)纠正来自PK动物的iPS细胞中的PK突变(3)并将纠正后的iPS细胞分化为移植后纠正疾病的造血干细胞。这些大型动物研究对于证明基于iPS的基因治疗遗传性贫血的有效性和安全性至关重要。两个核心将促进上述研究。Timothy Wick博士将领导一个生物反应器核心，用于大规模培养和将iPS细胞分化为造血祖细胞，Larry Lamb博士将领导一个造血细胞处理核心，用于从校正后的iPS细胞中纯化造血祖细胞。这两个核心对于将这项工作转化为人类临床试验至关重要。

DESCRIPTION (provided by applicant): The UAB Stem Cell Institute, the UAB BioMatrix Engineering and Regenerative Medicine (BERM) Center, and the UAB Hematopoietic Stem Cell Processing Facility will collaborate in a program to develop gene replacement therapy using induced pluripotent stem (iPS) cells for treatment of sickle cell disease. The program consists of two projects and two cores. In Project 1 (Human iPS Cells for Sickle Gene Replacement), Dr. Townes proposes to translate recent success in correcting sickle cell disease in a humanized sickle mouse model into human cells. The specific aims of the project are to: (1) produce human iPS cells from skin biopsy samples of patients with sickle cell disease (2) to correct the sickle mutation in iPS cells derived from patients (3) and to differentiate corrected iPS cells into transplantable hematopoietic stem cells that produce normal erythroid cells. In Project 2 (Canine iPS Cells For Pyruvate Kinase Gene Replacement), Dr. Clint Lothrop proposes to correct a large animal model of hereditary anemia (pyruvate kinase deficiency in dogs) by gene replacement in canine iPS cells. The specific aims of this project are: (1) to produce canine iPS cells from skin biopsy samples of dogs with hereditary pyruvate kinase (PK) deficiency (2) to correct the PK mutation in iPS cells derived from PK animals (3) and to differentiate corrected iPS cells into hematopoietic stem cells that correct the disease after transplantation. These large animal studies will be crucial to prove the efficacy and safety of iPS based gene therapy for hereditary anemias. Two cores will facilitate the studies described above. Dr. Timothy Wick will direct a Bioreactor Core for the large-scale culture and differentiation of iPS cells into hematopoietic progenitors, and Dr. Larry Lamb will direct a Hematopoietic Cell Processing Core for the purification of hematopoietic progenitors from corrected iPS cells. These two cores will be crucial to the translation of this work to human clinical trials.