Exploring Alternative iPS Cell Therapies for Recessive Dystrophic Epidermolysis Bullosa

项目摘要 没有有效的治疗方法可用于大疱性表皮病（EB），一组罕见的遗传性皮肤 这些疾病可能是毁灭性的，在某些情况下是致命的。重新编程成人皮肤细胞的能力 诱导多能干细胞（iPSC）现在提供了开发永久性矫正 治疗EB。我们在开发更安全，更有效的重编程，基因编辑和iPSC方面的进展 分化方案使我们能够建立“EB iPS细胞联盟”， 科罗拉多（Roop、Bilousova、Kogut和Bruckner博士）、斯坦福大学（由Oro博士领导）和哥伦比亚 大学（由Christiano博士领导）。该联盟目前正在开发一种基于iPSC的治疗方法， EB的严重隐性营养不良形式（RDEB），由基因校正的表皮片组成， 复合皮肤移植。目前的申请将探索两种新的iPSC疗法用于RDEB，以及非iPSC疗法。 目前赞助联盟、EB研究伙伴关系、EB医疗基金会的利润基金会 研究基金会和治愈EB慈善机构已同意提供所需的1对1匹配资金， 这份RFA申请。虽然皮肤移植可能是证明安全性和有效性的最快途径 对于RDEB患者，基于iPSC的治疗，从基因产生这些移植物所需的时间 校正的iPSC是冗长的，因此是昂贵的。因此，我们建议评估一种“喷雾- Avita Medical开发的“皮肤”递送系统，用于递送从基因编辑的 RDEB iPSC是皮肤移植的更直接的替代品。如果成功的话，“皮肤喷雾” 系统将减少患者应用的时间，而不是表皮生长所需的时间和成本， 复合移植物，并可能产生上级结果为EB患者由于较低的风险， 炎症和疤痕。此外，虽然基因校正的iPSC衍生的角质形成细胞可以校正 由于RDEB患者的皮肤表型，这些细胞将不能有效治疗严重的胃肠道疾病， 与RDEB相关的临床表现，通常需要使用饲管。因此，我们也将 评估基因校正的iPSC衍生的间充质干细胞（MSC）的全身递送是否可以促进 内部上皮和皮肤的伤口愈合。如果有效，MSC的全身递送不仅 彻底改变了我们治疗EB患者的方式，但也可能代表了一种治疗许多系统性疾病的新方法。 影响内部器官和组织的多灶性疾病和损伤。提出了以下目标： 将在符合cGMP的条件下产生基因校正的iPSC衍生的成纤维细胞和MSC。目标2将 探索使用“皮肤喷雾”装置递送基因校正的iPSC衍生的角质形成细胞的可行性 和成纤维细胞。目的3将评估全身递送的MSC改善创伤愈合的能力。 皮肤和内部上皮。目标4将为FDA提供初步的安全性和有效性数据。

Project Summary No effective treatments are available for epidermolysis bullosa (EB), a group of rare inherited skin blistering disorders that can be devastating, and in some cases lethal. The ability to reprogram adult skin cells into induced pluripotent stem cells (iPSC) now offers the possibility of developing a permanent corrective therapy for EB. Our advances in developing safer and more efficient reprogramming, gene editing and iPSC differentiation protocols allowed us to establish the “EB iPS Cell Consortium” comprised of the University of Colorado (Drs. Roop, Bilousova, Kogut and Bruckner), Stanford University (led by Dr. Oro) and Columbia University (led by Dr. Christiano). The Consortium is currently developing an iPSC-based therapy for the severe recessive dystrophic form of EB (RDEB) that is comprised of gene-corrected epidermal sheets and composite skin grafts. The current application will explore two novel iPSC therapies for RDEB, and the non- profit foundations that currently sponsor the Consortium, the EB Research Partnership, the EB Medical Research Foundation and the Cure EB Charity, have agreed to provide the required 1 to 1 matching funds for this RFA submission. Although skin grafts may be the fastest path forward to demonstrate safety and efficacy of an iPSC-based therapy for RDEB patients, the time required to generate these grafts from genetically corrected iPSCs is lengthy and consequently expensive. Therefore, we are proposing to evaluate a “spray-on- skin” delivery system developed by Avita Medical, for delivering skin cells differentiated from gene-edited RDEB iPSCs as a more straightforward alternative to skin grafts. If successful, the “spray-on-skin” delivery system would decrease the time to patient application vs. the time and cost it takes to grow epidermal and composite grafts and would potentially produce superior outcomes for EB patients due to the lower risk of inflammation and scarring. In addition, while gene-corrected iPSC-derived keratinocytes may correct the cutaneous phenotype in RDEB patients, these cells will not be effective in treating the severe gastrointestinal manifestations associated with RDEB, which often require the use of feeding tubes. Therefore, we will also assess if the systemic delivery of gene-corrected-iPSC-derived mesenchymal stem cells (MSCs) can facilitate wound healing in both internal epithelia and the skin. If effective, the systemic delivery of MSCs would not only revolutionize how we treat EB patients, but also potentially represent a novel approach to treat many systemic multifocal diseases and injuries that affect internal organs and tissues. The following aims are proposed: Aim 1 will generate gene-corrected iPSC-derived fibroblasts and MSCs under cGMP-compliant conditions. Aim 2 will explore the feasibility of using the “spray on skin” device to deliver gene-corrected iPSC-derived keratinocytes and fibroblasts. Aim 3 will assess the ability of systemically delivered MSCs to improve wound healing in the skin and internal epithelia. Aim 4 will generate preliminary safety and efficacy data for the FDA.