Transgene-Free Disease-Specific Induced Pluripotent Stem Cells from Patients with Type 1 and Type 2 Diabetes
Transgene-Free Disease-Specific Induced Pluripotent Stem Cells from Patients with Type 1 and Type 2 Diabetes
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DOI：
10.5966/sctm.2011-0044
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发表时间：
2012-06-01
影响因子：
6
通讯作者：
Ikeda, Yasuhiro
中科院分区：
文献类型：
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作者：
Kudva, Yogish C.;Ohmine, Seiga;Ikeda, Yasuhiro
诱导多能干细胞（iPSC）技术使得能够在不使用胚胎细胞来源的情况下从成体细胞衍生患者特异性多能干细胞。将来自糖尿病患者的iPSC再分化为胰岛将允许患者特异性疾病建模和用于衰竭胰岛的自体细胞替代疗法。迄今为止，已使用整合逆转录病毒载体从1型糖尿病患者中产生了糖尿病特异性iPSC。然而，载体整合到宿主基因组中可能会损害衍生的iPSC的生物安全性和分化倾向。尽管已经描述了各种无整合重编程系统，但它们重编程患者体细胞的效用在很大程度上仍未确定。在这里，我们使用非整合仙台病毒载体重新编程1型和2型糖尿病（T2 D）患者的细胞。仙台载体感染导致从糖尿病患者（包括一名85岁的T2 D患者）中可重复地产生无基因组修饰的iPSC（SV-iPSC）。SV-iPSC在8-12代内失去仙台病毒基因组和抗原，同时保持多能性。SV-iPSCs的全基因组转录组分析显示，内源性多能性基因的诱导和参与氧化应激反应和INK 4/ARF途径的基因的下调，包括p16（INK 4a），p15（INK 4 b）和p21（CIP 1）。SV-iPSC和用整合慢病毒载体制备的iPSC在全局基因表达谱中表现出显著的相似性。因此，仙台载体系统有助于将患者细胞可靠地重编程为无转基因的iPSC，为糖尿病和糖尿病相关并发症的个性化诊断和治疗方法提供多能平台。干细胞转化医学2012;1：451-461
The induced pluripotent stem cell (iPSC) technology enables derivation of patient-specific pluripotent stem cells from adult somatic cells without using an embryonic cell source. Redifferentiation of iPSCs from diabetic patients into pancreatic islets will allow patient-specific disease modeling and autologous cell replacement therapy for failing islets. To date, diabetes-specific iPSCs have been generated from patients with type 1 diabetes using integrating retroviral vectors. However, vector integration into the host genome could compromise the biosafety and differentiation propensities of derived iPSCs. Although various integration-free reprogramming systems have been described, their utility to reprogram somatic cells from patients remains largely undetermined. Here, we used nonintegrating Sendai viral vectors to reprogram cells from patients with type 1 and type 2 diabetes (T2D). Sendai vector infection led to reproducible generation of genomic modification-free iPSCs (SV-iPSCs) from patients with diabetes, including an 85-year-old individual with T2D. SV-iPSCs lost the Sendai viral genome and antigens within 8-12 passages while maintaining pluripotency. Genome-wide transcriptome analysis of SV-iPSCs revealed induction of endogenous pluripotency genes and downregulation of genes involved in the oxidative stress response and the INK4/ARF pathways, including p16(INK4a), p15(INK4b), and p21(CIP1). SV-iPSCs and iPSCs made with integrating lentiviral vectors demonstrated remarkable similarities in global gene expression profiles. Thus, the Sendai vector system facilitates reliable reprogramming of patient cells into transgene-free iPSCs, providing a pluripotent platform for personalized diagnostic and therapeutic approaches for diabetes and diabetes-associated complications. STEM CELLS TRANSLATIONAL MEDICINE 2012;1:451-461