Division of Endocrinology, Mayo Clinic, Rochester, MN 55905, USA.
Stem Cells Transl Med. 2012 Jun;1(6):451-61. doi: 10.5966/sctm.2011-0044. Epub 2012 May 30.
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.
诱导多能干细胞（iPSC）技术使人们能够从成人体细胞中获得患者特异性多能干细胞，而无需使用胚胎细胞来源。将糖尿病患者的 iPSC 重新分化为胰岛，将允许对患者特异性疾病进行建模，并对衰竭的胰岛进行自体细胞替代治疗。迄今为止，已经使用整合型逆转录病毒载体从 1 型糖尿病患者中生成了糖尿病特异性 iPSC。然而，载体整合到宿主基因组中可能会损害衍生 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，但它们在将体细胞从患者中重编程的实用性在很大程度上仍未确定。在这里，我们使用非整合性的 Sendai 病毒载体来重编程 1 型和 2 型糖尿病（T2D）患者的细胞。Sendai 病毒载体感染可从糖尿病患者中产生可重复的无基因组修饰的 iPSC（SV-iPSC），包括一名患有 T2D 的 85 岁个体。SV-iPSC 在 8-12 个传代过程中失去了 Sendai 病毒基因组和抗原，同时保持多能性。SV-iPSC 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 INK4/ARF 途径的基因下调，包括 p16(INK4a)、p15(INK4b)和 p21(CIP1)。SV-iPSC 和使用整合性慢病毒载体生成的 iPSC 在全基因组表达谱方面表现出显著的相似性。因此，Sendai 病毒载体系统可促进将患者细胞可靠地重编程为无转基因的 iPSC，为糖尿病及其相关并发症的个性化诊断和治疗方法提供了一个多能性平台。
