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.

项目概要 大疱性表皮松解症（EB）是一种罕见的遗传性水疱，目前尚无有效的治疗方法 可能是毁灭性的，在某些情况下甚至是致命的疾病 技术的突破使成人成为可能。 皮肤细胞被重新编程为诱导多能干细胞（iPSC）现在提供了可能性 开发 EB 的永久性纠正疗法。治疗方法依赖于 EB 的产生。 患者特异性 iPSC，然后进行基因校正并分化为适合的皮肤细胞 从同一位需要治疗的患者身上产生 iPSC 不仅有可能。 避免免疫排斥的并发症，同时也提供无限且可扩展的患者来源 对于由母体 R01 资助的初始研究，我们重点关注适合移植的特定细胞。 为一群患有 EB Dowling-Meara 亚型的患者开发基于 iPSC 的疗法 simplex (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 案例 这项 EBS-DM 研究使我们能够解决所有主要问题。 开发临床相关的基于 iPSC 的 EBS 疗法所需的障碍和先决条件。 具体来说，我们开发了一种新颖、有效的方法来生成临床相关的整合 - 我们还使用非整合选择标记基因编辑技术成功地获得了免费的 iPSC。 纠正 EBS 患者产生的 iPSC 的遗传缺陷。 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起来 这一修订申请提议扩展一步组合基因编辑和 重编程方法治疗更严重的 EB，即隐性营养不良 EB (RDEB)，并推进我们的研究 鉴于需要多个小组之间的合作努力才能获得临床试验的结果。 确保将基于 iPSC 的复杂疗法成功转化为临床，我们最近进入 与斯坦福大学和哥伦比亚大学组成联盟，以促进最佳的开发 用于 RDEB 治疗的基因校正 iPSC 衍生细胞产品的制造方案。 该提案的目标 1 是提高基于 iPSC 的安全性并降低制造复杂性 通过将重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 生产符合cGMP标准的试剂，分别用于重编程和基因编辑，以协助 联盟正在生成研究性新药 (IND) - 为 FDA 提供数据。 联盟成功获得 RDEB 临床试验批准将扩大其治疗潜力 iPSC 用于治疗影响其他内脏器官的各种单基因疾病，其中 难以监测基于 iPSC 的疗法的不良反应将使它们不太可能成为首选目标。