Testing the Therapeutic Potential of iPS Cells for Inherited Skin Diseases

测试 iPS 细胞对遗传性皮肤病的治疗潜力

• 批准号: 9516699
• 负责人: Dennis Roop
• 金额: $ 42.89万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-17 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9516699
• 关键词: Address; Adopted; Adult; Adverse effects; Affect; Award; Back; Biomanufacturing; Biopsy; Bulla; CRISPR/Cas technology; Cell Therapy; Cells; Cessation of life; Cicatrix; Clinic; Clinical; Clinical Data; Clinical Trials; Collagen Type VII; Colorado; Communication; Complex; Complication; Data; Development; Disease; Ensure; Epidermolysis Bullosa; Epidermolysis Bullosa Dystrophica; Epidermolysis Bullosa Simplex; Fibroblasts; Funding; Future; Generations; Genes; Genetic; Goals; Hereditary Disease; Hot Spot; Human Resources; Immune; Impaired wound healing; Incidence; Inherited; Investigational Drugs; Investigational New Drug Application; Keratin; Mendelian disorder; Messenger RNA; Methods; Monitor; Mutation; One-Step dentin bonding system; Organ; Other Genetics; Parents; Patients; Phase I Clinical Trials; Procedures; Process; Production; Proteins; Protocols documentation; Quality Control; RNA; Reagent; Research; Safety; Severities; Skin; Source; Stem cells; Techniques; Technology; Testing; Therapeutic; Translations; Transplantation; Universities; Work; base; clinical development; clinical translation; clinically relevant; cohort; cost; effective therapy; gene correction; improved; induced pluripotent stem cell; keratinocyte; member; novel; parent grant; pre-clinical; pre-clinical research; restoration; risk benefit ratio; skin disorder; success; therapeutic evaluation; therapy development

Project Summary No effective treatments are available for epidermolysis bullosa (EB), a group of rare inherited blistering disorders that can be devastating and in some cases lethal. The technological breakthrough that allows adult skin cells to be reprogrammed into induced pluripotent stem cells (iPSCs) now offers the possibility of developing a permanent corrective therapy for EB. The therapeutic approach relies on the generation of patient-specific iPSCs, which then undergo genetic correction and differentiation into skin cells suitable for transplantation. The generation of iPSCs from the same patient in need of treatment would not only potentially avoid the complication of immune rejection, but also provide an unlimited and scalable source of patient- specific cells suitable for transplantation. For the initial studies funded by the parent R01, we focused on developing an iPSC-based therapy for a cohort of patients suffering from the Dowling-Meara subtype of EB simplex (EBS-DM), who share the same R125C “hot spot mutation” in the keratin 14 (Krt14) protein that is responsible for 70% of the EBS-DM cases. This EBS-DM study has allowed us to address all of the major obstacles and pre-requisites required for the development of a clinically relevant iPSC-based therapy for EBS. Specifically, we have developed a novel, efficient method for the generation of clinically relevant integration- free iPSCs. We have also used non-integrating selection marker-free gene editing technology to successfully correct the genetic defect in iPSCs generated from an EBS patient. In addition, our high-efficiency of reprogramming has recently allowed us to combine gene editing and reprogramming of EBS fibroblasts into a one-step procedure. This revision application is proposing to expand the one-step combined gene editing and reprogramming approach to a more severe form of EB, recessive dystrophic EB (RDEB) and to advance our findings toward a clinical trial. Given that the collaborative efforts among several groups will be required to ensure the successful translation of complex iPSC-based therapies into the clinic, we have recently entered into a consortium with Stanford and Columbia Universities to facilitate the development of an optimal manufacturing protocol for genetically corrected iPSC-derived cell products for RDEB treatment. Therefore, Aim 1 of this proposal is to improve the safety and reduce the manufacturing complexity of an iPSC-based therapy for RDEB by combining reprogramming and gene editing into a one-step procedure; Aims 2 and 3 are to manufacture cGMP-compliant reagents for reprogramming and gene editing respectively to assist the Consortium in generating Investigational New Drug (IND) - enabling data for the FDA. The EB iPS Cell Consortium’s success in obtaining approval for a clinical trial for RDEB would expand the therapeutic potential of iPSCs for the treatment of a variety of monogenic diseases affecting other internal organs, where the difficulty in monitoring adverse effects of an iPSC-based therapy would make them unlikely first targets.

项目摘要 大疱性表皮松解症(EB)是一种罕见的遗传性水泡，目前尚无有效的治疗方法 疾病可能是毁灭性的，在某些情况下是致命的。让成年人能够 皮肤细胞被重新编程为诱导多能干细胞(IPSCs)现在提供了一种可能性 开发一种针对EB的永久性矫正疗法。治疗方法依赖于产生 特定于患者的IPSCs，然后经过基因校正和分化成适合于 移植。从同一个需要治疗的患者身上产生ipscs不仅有可能 避免了免疫排斥的并发症，但也提供了无限和可扩展的患者来源- 适合移植的特定细胞。对于由母公司R01资助的初步研究，我们专注于 为一组患有Dowling-Meara亚型EB的患者开发基于IPSC的疗法 单纯型(EBS-DM)，在角蛋白14(Krt14)蛋白中具有相同的R125C“热点突变”，即 负责70%的EBS-DM病例。这项EBS-DM研究使我们能够解决所有主要的 开发临床相关的以IPSC为基础的治疗EBS所需的障碍和先决条件。 具体地说，我们已经开发了一种新的、高效的方法来生成临床相关的整合- 免费的iPSCs。我们还利用非整合选择无标记基因编辑技术成功地 纠正一名EBS患者产生的IPSCs的基因缺陷。另外，我们的高效率的 重新编程最近允许我们将基因编辑和EBS成纤维细胞的重新编程结合在一起 一步法。这一修订申请建议扩大一步结合基因编辑和 对一种更严重的EB，隐性营养不良性EB(RDEB)的重新编程方法，并推进我们的 临床试验的结果。鉴于几个小组之间的合作努力将需要 确保基于IPSC的复杂疗法成功转化为临床，我们最近进入了 与斯坦福大学和哥伦比亚大学结成联盟，以促进开发最佳 用于RDEB治疗的转基因IPSC衍生细胞产品的制造方案。因此， 该提案的目标1是提高基于IPSC的IPSC的安全性和制造复杂性 通过将重新编程和基因编辑结合到一步程序中来治疗RDEB；目标2和3是 制造符合cGMP的试剂，分别用于重新编程和基因编辑，以帮助 为FDA生成研究新药(IND)使能数据的联盟。EB iPS细胞 该财团成功获得RDEB临床试验的批准，将扩大治疗潜力 用于治疗各种影响其他内脏器官的单基因疾病，其中 很难监测基于ipsc的疗法的不良反应，这将使他们不太可能成为第一个目标。