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 没有有效的治疗方法可用于表皮溶解球状（EB），这是一组罕见的遗传泡沫 可能是毁灭性的疾病，在某些情况下是致命的。允许成人的技术突破 皮肤细胞要重新编程为诱导的多能干细胞（IPSC），现在提供了 为EB开发永久性纠正疗法。治疗方法依赖于产生 患者特异性的IPSC，然后经过遗传校正和分化为皮肤细胞 移植。需要治疗的同一患者的IPSC不仅可能有潜在地 避免免疫排斥的并发症，但也提供了无限且可扩展的患者来源 适合移植的特定细胞。对于由父r01资助的最初研究，我们专注于 开发基于IPSC的疗法，用于患有EB的Dowling-Meara亚型的患者队列 单纯形（EBS-DM），他在角蛋白14（KRT14）蛋白中共享相同的R125C“热点突变” 负责70％的EBS-DM病例。这项EBS-DM研究使我们能够解决所有专业 开发基于IPSC的EBS疗法所需的障碍和先决条件。 具体而言，我们已经开发了一种新型，有效的方法来生成临床相关的整合 - 免费的IPSC。我们还使用了非整合选择的无标记基因编辑技术来成功 纠正从EBS患者产生的IPSC中的遗传缺陷。此外，我们的高效 重编程最近使我们能够将EBS成纤维细胞的基因编辑和重编程结合到一个 一步过程。此修订应用程序提议扩展一步组合基因编辑和 重新编程方法更严重的EB，隐性营养不良EB（RDEB），并促进我们的 临床试验的发现。鉴于需要几个小组之间的协作努力 确保将复杂的基于IPSC的疗法成功翻译为诊所，我们最近进入了 与斯坦福大学和哥伦比亚大学一起进入一个财团，以促进最佳的发展 用于遗传校正的IPSC衍生细胞产品的制造方案用于RDEB治疗。所以， 该提案的目标1是提高安全性并降低基于IPSC的制造复杂性 通过将重编程和基因编辑结合到一步程序中，用于RDEB的治疗；目标2和3是 制造符合CGMP的试剂，分别用于重编程和基因编辑以帮助 产生研究新药（IND）的联盟 - 启用FDA的数据。 EB IPS单元格 财团在获得RDEB临床试验批准方面的成功将扩大治疗潜力 IPSC的治疗，以治疗影响其他内部器官的多种单基因疾病 在监测基于IPSC的治疗的不良影响方面很难使它们不太可能成为第一个目标。