我们推出营养不良性大疱性表皮松解症细胞疗法 (DEBCT)，这是一个可扩展的平台，可生产用于根治性治疗的自体器官型 iPS 细胞衍生的诱导皮肤复合材料 (iSC) 移植物。临床级制造将 CRISPR 介导的基因校正与重编程整合为一步，加速从患者体内衍生出经过 COL7A1 编辑的 iPS 细胞。分化为表皮、真皮和黑素细胞祖细胞后，CD49f 富集，最大限度地减少成熟异质性。来自四名具有不同突变的患者的 iSC 的小鼠异种移植在 1 个月时表现出疾病缓解活性。新一代测序、生物分布和致瘤性测定在 1-9 个月内建立了良好的安全性。单细胞转录组学揭示 iSC 由主要皮肤细胞谱系组成，包括突出的全克隆干细胞样角质形成细胞特征，以及最近描述的成纤维细胞的 Gibbin 依赖性特征。后者与 iSC 的可移植性增强相关。总之，DEBCT 克服了制造和安全障碍，并建立了一种可重复、安全且符合 cGMP 的治疗方法来治愈 DEB 患者的病变。© 2024。作者。

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.© 2024. The Author(s).