Phondeechareon Tanapol, Wattanapanitch Methichit, U-Pratya Yaowalak, Damkham Chanapa, Klincumhom Nuttha, Lorthongpanich Chanchao, Kheolamai Pakpoom, Laowtammathron Chuti, Issaragrisil Surapol
Siriraj Center of Excellence for Stem Cell Research, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Ann Hematol. 2016 Oct;95(10):1617-25. doi: 10.1007/s00277-016-2756-1. Epub 2016 Jul 28.
Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired hemolytic anemia caused by lack of CD55 and CD59 on blood cell membrane leading to increased sensitivity of blood cells to complement. Hematopoietic stem cell transplantation (HSCT) is the only curative therapy for PNH, however, lack of HLA-matched donors and post-transplant complications are major concerns. Induced pluripotent stem cells (iPSCs) derived from patients are an attractive source for generating autologous HSCs to avoid adverse effects resulting from allogeneic HSCT. The disease involves only HSCs and their progeny; therefore, other tissues are not affected by the mutation and may be used to produce disease-free autologous HSCs. This study aimed to derive PNH patient-specific iPSCs from human dermal fibroblasts (HDFs), characterize and differentiate to hematopoietic cells using a feeder-free protocol. Analysis of CD55 and CD59 expression was performed before and after reprogramming, and hematopoietic differentiation. Patients' dermal fibroblasts expressed CD55 and CD59 at normal levels and the normal expression remained after reprogramming. The iPSCs derived from PNH patients had typical pluripotent properties and differentiation capacities with normal karyotype. After hematopoietic differentiation, the differentiated cells expressed early hematopoietic markers (CD34 and CD43) with normal CD59 expression. The iPSCs derived from HDFs of PNH patients have normal levels of CD55 and CD59 expression and hold promise as a potential source of HSCs for autologous transplantation to cure PNH patients.
阵发性睡眠性血红蛋白尿症（PNH）是一种获得性溶血性贫血，由血细胞细胞膜上缺乏CD55和CD59导致血细胞对补体的敏感性增加引起。造血干细胞移植（HSCT）是PNH的唯一治愈性疗法，然而，缺乏HLA匹配的供体和移植后并发症是主要问题。源自患者的诱导多能干细胞（iPSC）是产生自体造血干细胞的有吸引力的来源，以避免异基因HSCT产生的不良反应。该疾病仅涉及造血干细胞及其后代；因此，其他组织不受突变影响，可用于产生无病的自体造血干细胞。本研究旨在从人皮肤成纤维细胞（HDF）中获得PNH患者特异性iPSC，使用无饲养层方案进行表征并分化为造血细胞。在重编程前后以及造血分化前后进行CD55和CD59表达分析。患者的皮肤成纤维细胞正常表达CD55和CD59，重编程后仍保持正常表达。源自PNH患者的iPSC具有典型的多能性特性和分化能力，核型正常。造血分化后，分化细胞表达早期造血标志物（CD34和CD43），CD59表达正常。源自PNH患者HDF的iPSC具有正常水平的CD55和CD59表达，有望作为自体移植治疗PNH患者的潜在造血干细胞来源。
