Division of Immunology, Children's Hospital Boston, Harvard Medical School, Boston, MA, USA.
J Allergy Clin Immunol. 2011 Jun;127(6):1400-7.e4. doi: 10.1016/j.jaci.2010.11.008. Epub 2010 Dec 24.
The novel ability to epigenetically reprogram somatic cells into induced pluripotent stem cells (iPSCs) through the exogenous expression of transcription promises to revolutionize the study of human diseases.
Here we report on the generation of 25 iPSC lines from 6 patients with various forms of primary immunodeficiencies (PIDs) affecting adaptive immunity, innate immunity, or both.
Patients' dermal fibroblasts were reprogrammed by expression of 4 transcription factors, octamer-binding transcription factor 4 (OCT4), sex determining region Y-box 2 (SOX2), Krueppel-like factor 4 (KLF4), and cellular myelomonocytosis proto-oncogene (cMYC), by using a single excisable polycistronic lentiviral vector.
iPSCs derived from patients with PIDs show a stemness profile that is comparable with that observed in human embryonic stem cells. After in vitro differentiation into embryoid bodies, pluripotency of the patient-derived iPSC lines was demonstrated by expression of genes characteristic of each of the 3 embryonic layers. We have confirmed the patient-specific origin of the iPSC lines and ascertained maintenance of karyotypic integrity.
By providing a limitless source of diseased stem cells that can be differentiated into various cell types in vitro, the repository of iPSC lines from patients with PIDs represents a unique resource to investigate the pathophysiology of hematopoietic and extrahematopoietic manifestations of these diseases and might assist in the development of novel therapeutic approaches based on gene correction.
通过外源性表达转录因子，体细胞可以被重编程为诱导多能干细胞（iPSCs），这种新的能力有望彻底改变人类疾病的研究。
本研究报告了从 6 名患有各种原发性免疫缺陷（PID）的患者中生成 25 条 iPSC 系，这些 PID 影响适应性免疫、固有免疫或两者兼有。
通过表达 4 种转录因子（八聚体结合转录因子 4（OCT4）、性别决定区 Y 框 2（SOX2）、Krüppel 样因子 4（KLF4）和细胞髓系原癌基因（cMYC）），使用单一可切除的多顺反子慢病毒载体，将患者的真皮成纤维细胞重编程为 iPSC。
从 PID 患者中获得的 iPSC 显示出与人类胚胎干细胞相似的干性特征。在体外分化为类胚体后，通过表达各 3 个胚层特征基因，证明了患者来源的 iPSC 系的多能性。我们已经证实了 iPSC 系的患者特异性起源，并确定了染色体组型完整性得以维持。
通过提供无限来源的患病干细胞，这些细胞可以在体外分化为各种细胞类型，来自 PID 患者的 iPSC 系库代表了一个独特的资源，可以用于研究这些疾病的造血和非造血表现的病理生理学，并且可能有助于基于基因纠正的新型治疗方法的开发。
