Abstract：
BACKGROUND: No efficient treatment has yet been established for epidermolytic ichthyosis (EI), which is caused by pathogenic variants of KRT1 or KRT10. Patients with ichthyosis with confetti (IWC) have multiple normal-appearing spots, caused by the revertant somatic recombination of pathogenic variants that occurs at each spot independently. Additionally, some patients with EI have large areas of normal skin due to revertant postzygotic mosaicism.
OBJECTIVE: To assess the feasibility of transplanting cultured epidermal autografts (CEAs) produced from revertant epidermal keratinocytes in patients with EI and IWC.
METHODS: We performed a clinical trial of treatment with CEAs produced from each patient\'s own revertant epidermal keratinocytes as a proof-of-concept study. This was a single-arm, open, unmasked, uncontrolled, single-assignment, treatment-purpose study. The primary outcome was the percentage area that lacked recurrence of ichthyosis lesions 4 weeks after the final transplant. The secondary outcome was the percentage area lacking recurrence of ichthyosis lesions 24 weeks after the initial transplantation. The trial was registered with the Japan Registry of Clinical Trials (jRCTb041190097).
RESULTS: We successfully produced CEAs from genetically confirmed revertant skin from two patients with mosaic EI and from one patient with IWC and confirmed by amplicon sequencing and droplet digital polymerase chain reaction analysis that the CEAs mainly consisted of revertant wild-type cells. Single-cell RNA sequencing analysis confirmed the normal proliferation and safety profiling of CEAs. CEAs were transplanted onto desquamated lesional sites in the patients. Four weeks post-transplantation, the percentage area lacking recurrence of ichthyosis lesions in the three patients was 40%, 100% and 100% respectively, although recurrence of ichthyosis lesions was seen at the site of CEA transplantation in all three patients at 24 weeks post-transplantation.
CONCLUSIONS: CEAs from normal skin have the potential to be a safe and local treatment option for EI and IWC.
Epidermolytic ichthyosis is a rare skin condition that causes redness, blistering and thickening of the skin. There is currently no effective treatment for the disease, which is caused by mutations in the genes KRT1 or KRT10. People with a type of the disease called ‘ichthyosis with confetti’ have many normal-appearing spots that are caused by the natural repair of the gene mutations. Some people with epidermolytic ichthyosis have large areas of healthy skin as a result of genetic mutations having been corrected. In this study, we successfully produced skin grafts from the healthy skin of two patients with epidermolytic ichthyosis and one with ‘ichthyosis with confetti’. We confirmed that the skin grafts mainly consisted of repaired skin cells. A technique called ‘single-cell RNA sequencing’ confirmed the skin cells in the skin grafts behaved like healthy skin cells and that the grafts were safe. Overall, our study findings suggest that skin grafts taken from skin consisting of genetically normal keratinocytes that have undergone self-repair have potential to be a safe treatment option for patients with severe epidermolytic ichthyosis and ‘ichthyosis with confetti’.
OBJECTIVE: To assess the feasibility of transplanting cultured epidermal autografts (CEAs) produced from revertant epidermal keratinocytes in patients with EI and IWC.
METHODS: We performed a clinical trial of treatment with CEAs produced from each patient\'s own revertant epidermal keratinocytes as a proof-of-concept study. This was a single-arm, open, unmasked, uncontrolled, single-assignment, treatment-purpose study. The primary outcome was the percentage area that lacked recurrence of ichthyosis lesions 4 weeks after the final transplant. The secondary outcome was the percentage area lacking recurrence of ichthyosis lesions 24 weeks after the initial transplantation. The trial was registered with the Japan Registry of Clinical Trials (jRCTb041190097).
RESULTS: We successfully produced CEAs from genetically confirmed revertant skin from two patients with mosaic EI and from one patient with IWC and confirmed by amplicon sequencing and droplet digital polymerase chain reaction analysis that the CEAs mainly consisted of revertant wild-type cells. Single-cell RNA sequencing analysis confirmed the normal proliferation and safety profiling of CEAs. CEAs were transplanted onto desquamated lesional sites in the patients. Four weeks post-transplantation, the percentage area lacking recurrence of ichthyosis lesions in the three patients was 40%, 100% and 100% respectively, although recurrence of ichthyosis lesions was seen at the site of CEA transplantation in all three patients at 24 weeks post-transplantation.
CONCLUSIONS: CEAs from normal skin have the potential to be a safe and local treatment option for EI and IWC.
Epidermolytic ichthyosis is a rare skin condition that causes redness, blistering and thickening of the skin. There is currently no effective treatment for the disease, which is caused by mutations in the genes KRT1 or KRT10. People with a type of the disease called ‘ichthyosis with confetti’ have many normal-appearing spots that are caused by the natural repair of the gene mutations. Some people with epidermolytic ichthyosis have large areas of healthy skin as a result of genetic mutations having been corrected. In this study, we successfully produced skin grafts from the healthy skin of two patients with epidermolytic ichthyosis and one with ‘ichthyosis with confetti’. We confirmed that the skin grafts mainly consisted of repaired skin cells. A technique called ‘single-cell RNA sequencing’ confirmed the skin cells in the skin grafts behaved like healthy skin cells and that the grafts were safe. Overall, our study findings suggest that skin grafts taken from skin consisting of genetically normal keratinocytes that have undergone self-repair have potential to be a safe treatment option for patients with severe epidermolytic ichthyosis and ‘ichthyosis with confetti’.
摘要：
背景：对于由KRT1或KRT10致病变异引起的表皮性鱼鳞病（EI），尚未建立有效的治疗方法。患有五彩纸屑的鱼鳞病患者（IWC）显示多个正常出现的斑点，由独立发生在每个斑点的致病变体的回复体重组引起的。此外,一些EI患者由于回复性合子后镶嵌性而有大面积的正常皮肤。
目的：评估在EI和IWC患者中移植由逆转表皮角质形成细胞产生的培养的表皮自体移植物（CEA）的可行性。
方法：我们进行了一项临床试验，对每个患者自身的回复体表皮角质形成细胞产生的CEA进行治疗，作为概念验证研究。这是单臂,打开（不使用掩蔽），不受控制,单一分配，治疗目的研究。主要结果是最终移植后4周鱼鳞病病灶无复发区域的发生率（％）。次要结果是初次移植后24周无鱼鳞病病灶复发的面积率（％）。
结果：我们成功地从两名镶嵌EI患者和一名IWC患者的基因证实的回复体皮肤中生产了CEA，并通过扩增子测序和液滴数字PCR分析，遗传证实了CEA主要由回复体野生型细胞组成。单细胞RNA测序分析证实了CEA的正常增殖和安全性。将CEA移植到患者脱皮的病变部位。移植4周后,三例鱼鳞病病灶无复发区域的发生率为39.52%,100.0%,和100.0%，尽管3例患者在移植后24周均在CEA移植部位发现鱼鳞病病灶复发。
结论：来自正常皮肤的CEAs有可能成为EI和IWC的安全和局部治疗选择。
背景：jRCTb041190097。
目的：评估在EI和IWC患者中移植由逆转表皮角质形成细胞产生的培养的表皮自体移植物（CEA）的可行性。
方法：我们进行了一项临床试验，对每个患者自身的回复体表皮角质形成细胞产生的CEA进行治疗，作为概念验证研究。这是单臂,打开（不使用掩蔽），不受控制,单一分配，治疗目的研究。主要结果是最终移植后4周鱼鳞病病灶无复发区域的发生率（％）。次要结果是初次移植后24周无鱼鳞病病灶复发的面积率（％）。
结果：我们成功地从两名镶嵌EI患者和一名IWC患者的基因证实的回复体皮肤中生产了CEA，并通过扩增子测序和液滴数字PCR分析，遗传证实了CEA主要由回复体野生型细胞组成。单细胞RNA测序分析证实了CEA的正常增殖和安全性。将CEA移植到患者脱皮的病变部位。移植4周后,三例鱼鳞病病灶无复发区域的发生率为39.52%,100.0%,和100.0%，尽管3例患者在移植后24周均在CEA移植部位发现鱼鳞病病灶复发。
结论：来自正常皮肤的CEAs有可能成为EI和IWC的安全和局部治疗选择。
背景：jRCTb041190097。
