PDF(Pubmed)
Abstract：
We present Dystrophic Epidermolysis Bullosa Cell Therapy (DEBCT), a scalable platform producing autologous organotypic iPS cell-derived induced skin composite (iSC) grafts for definitive treatment. Clinical-grade manufacturing integrates CRISPR-mediated genetic correction with reprogramming into one step, accelerating derivation of COL7A1-edited iPS cells from patients. Differentiation into epidermal, dermal and melanocyte progenitors is followed by CD49f-enrichment, minimizing maturation heterogeneity. Mouse xenografting of iSCs from four patients with different mutations demonstrates disease modifying activity at 1 month. Next-generation sequencing, biodistribution and tumorigenicity assays establish a favorable safety profile at 1-9 months. Single cell transcriptomics reveals that iSCs are composed of the major skin cell lineages and include prominent holoclone stem cell-like signatures of keratinocytes, and the recently described Gibbin-dependent signature of fibroblasts. The latter correlates with enhanced graftability of iSCs. In conclusion, DEBCT overcomes manufacturing and safety roadblocks and establishes a reproducible, safe, and cGMP-compatible therapeutic approach to heal lesions of DEB patients.
摘要：
我们介绍了营养不良性大疱性表皮松解症细胞疗法（DEBCT），一种可扩展的平台，可产生自体器官型iPS细胞衍生的诱导皮肤复合（iSC）移植物，用于最终治疗。临床级制造将CRISPR介导的基因校正与重编程整合到一个步骤中，加速从患者中衍生COL7A1编辑的iPS细胞。分化为表皮，真皮和黑素细胞祖细胞随后是CD49f富集，将成熟异质性降至最低。来自4名具有不同突变的患者的iSC的小鼠异种移植在1个月时显示疾病修饰活性。下一代测序，生物分布和致瘤性测定在1-9个月时建立了良好的安全性。单细胞转录组学显示，iSC由主要的皮肤细胞谱系组成，并包括角质形成细胞的突出的全克隆干细胞样特征，以及最近描述的成纤维细胞的Gibbin依赖性特征。后者与iSC的增强的可嫁接性相关。总之,DEBCT克服了制造和安全障碍，并建立了可重复的，安全,与cGMP相容的治疗方法可以治愈DEB患者的病变。
