Department of Molecular Medicine, College of Medicine, Mayo Clinic, Rochester, Minnesota 55905, USA.
Mol Ther. 2013 Jan;21(1):228-39. doi: 10.1038/mt.2012.245. Epub 2012 Nov 27.
Nuclear reprogramming of adult somatic tissue enables embryo-independent generation of autologous, patient-specific induced pluripotent stem (iPS) cells. Exploiting this emergent regenerative platform for individualized medicine applications requires the establishment of bioequivalence criteria across derived pluripotent lines and lineage-specified derivatives. Here, from individual patients with type 1 diabetes (T1D) multiple human iPS clones were produced and prospectively screened using a battery of developmental markers to assess respective differentiation propensity and proficiency in yielding functional insulin (INS)-producing progeny. Global gene expression profiles, pluripotency expression patterns, and the capacity to differentiate into SOX17- and FOXA2-positive definitive endoderm (DE)-like cells were comparable among individual iPS clones. However, notable intrapatient variation was evident upon further guided differentiation into HNF4α- and HNF1β-expressing primitive gut tube, and INS- and glucagon (GCG)-expressing islet-like cells. Differential dynamics of pluripotency-associated genes and pancreatic lineage-specifying genes underlined clonal variance. Successful generation of glucose-responsive INS-producing cells required silencing of stemness programs as well as the induction of stage-specific pancreatic transcription factors. Thus, comprehensive fingerprinting of individual clones is mandatory to secure homogenous pools amenable for diagnostic and therapeutic applications of iPS cells from patients with T1D.
成体体细胞的核重编程使胚胎独立产生自体、患者特异性诱导多能干细胞（iPS 细胞）成为可能。为了将这一新兴的再生平台应用于个体化医学，需要在衍生的多能系和谱系特异性衍生物之间建立生物等效性标准。在这里，从 1 型糖尿病（T1D）的个体患者中，产生了多个人类 iPS 克隆，并使用一系列发育标志物进行前瞻性筛选，以评估各自的分化倾向和产生功能性胰岛素（INS）产生后代的能力。个体 iPS 克隆之间的全球基因表达谱、多能性表达模式以及分化为 SOX17 和 FOXA2 阳性确定性内胚层（DE）样细胞的能力具有可比性。然而，在进一步指导分化为表达 HNF4α 和 HNF1β 的原始肠管以及表达 INS 和胰高血糖素（GCG）的胰岛样细胞时，明显存在个体内变异。多能性相关基因和胰腺谱系特异性基因的差异动力学突出了克隆变异性。成功生成葡萄糖反应性 INS 产生细胞需要沉默干性程序以及诱导阶段特异性胰腺转录因子。因此，对个体克隆进行全面指纹分析对于确保同质池可用于来自 T1D 患者的 iPS 细胞的诊断和治疗应用是必要的。
