Ohmine Seiga, Squillace Karen A, Hartjes Katherine A, Deeds Michael C, Armstrong Adam S, Thatava Tayaramma, Sakuma Toshie, Terzic Andre, Kudva Yogish, Ikeda Yasuhiro
Department of Molecular Medicine, Mayo Clinic, Rochester, MN 55905, USA.
Aging (Albany NY). 2012 Jan;4(1):60-73. doi: 10.18632/aging.100428.
Nuclear reprogramming enables patient-specific derivation of induced pluripotent stem (iPS) cells from adult tissue. Yet, iPS generation from patients with type 2 diabetes (T2D) has not been demonstrated. Here, we report reproducible iPS derivation of epidermal keratinocytes (HK) from elderly T2D patients. Transduced with human OCT4, SOX2, KLF4 and c-MYC stemness factors under serum-free and feeder-free conditions, reprogrammed cells underwent dedifferentiation with mitochondrial restructuring, induction of endogenous pluripotency genes - including NANOG, LIN28, and TERT, and down-regulation of cytoskeletal, MHC class I- and apoptosis-related genes. Notably, derived iPS clones acquired a rejuvenated state, characterized by elongated telomeres and suppressed senescence-related p15INK4b/p16INK4a gene expression and oxidative stress signaling. Stepwise guidance with lineage-specifying factors, including Indolactam V and GLP-1, redifferentiated HK-derived iPS clones into insulin-producing islet-like progeny. Thus, in elderly T2D patients, reprogramming of keratinocytes ensures a senescence-privileged status yielding iPS cells proficient for regenerative applications.
核重编程能够从成体组织中获得患者特异性的诱导多能干细胞（iPS细胞）。然而，尚未证实能够从2型糖尿病（T2D）患者中生成iPS细胞。在此，我们报告了从老年T2D患者的表皮角质形成细胞（HK）中可重复地获得iPS细胞。在无血清和无饲养层条件下，用人OCT4、SOX2、KLF4和c-MYC干性因子进行转导，重编程细胞经历去分化，伴有线粒体重塑、内源性多能性基因（包括NANOG、LIN28和TERT）的诱导，以及细胞骨架、MHC I类和凋亡相关基因的下调。值得注意的是，所获得的iPS克隆进入了年轻化状态，其特征为端粒延长、衰老相关的p15INK4b/p16INK4a基因表达受到抑制以及氧化应激信号传导被抑制。使用包括吲哚内酰胺V和胰高血糖素样肽-1在内的谱系特异性因子进行逐步引导，可将源自HK的iPS克隆重新分化为产生胰岛素的胰岛样后代。因此，在老年T2D患者中，角质形成细胞的重编程确保了一种衰老特权状态，从而产生适用于再生应用的iPS细胞。
