Engineering a glucose-responsive human insulin-secreting cell line from islets of langerhans isolated from a patient with persistent hyperinsulinemic hypoglycemia of infancy
Engineering a glucose-responsive human insulin-secreting cell line from islets of langerhans isolated from a patient with persistent hyperinsulinemic hypoglycemia of infancy
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DOI：
10.1074/jbc.274.48.34059
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发表时间：
1999-11-26
影响因子：
4.8
通讯作者：
Dunne, MJ
中科院分区：
文献类型：
--
作者：
MacFarlane, WM;Chapman, JC;Dunne, MJ
婴儿持续性高胰岛素性低血糖 (PHHI) 是一种新生儿疾病，其特征是胰岛素分泌失调并伴有严重低血糖。我们发现，从 PHHI 患者的胰腺中分离的胰岛细胞在培养物中增殖，同时保持 β 细胞样表型。PHHI 衍生细胞系 (NES2Y) 表现出源自这些患者的胰岛细胞典型的胰岛素分泌特征，即，它们没有K-ATP 通道活性，因此在没有葡萄糖的情况下分泌高水平的胰岛素。此外，它们表现出同源结构域转录因子 PDX1 的表达受损，PDX1 是连接营养代谢与胰岛素基因表达调节的信号通路的关键组成部分。为了修复这些缺陷，NES2Y 细胞用编码 K-ATP 通道的两种成分（SUR1 和 Kir6.2）和 PDX1 的 cDNA 三重转染，一种选定的克隆细胞系 (NISK9) 具有正常的 K-ATP 通道活性，并且由于细胞内 Ca2+ 稳态 ([Ca2+](i)) 的变化，在葡萄糖浓度的生理范围内分泌胰岛素。这种改造 PHHI 衍生胰岛细胞的方法可用于 PHHI 的基因治疗和用于施用胰岛素治疗糖尿病的细胞工程技术。
Persistent hyperinsulinemic hypoglycemia of infancy (PHHI) is a neonatal disease characterized by dysregulation of insulin secretion accompanied by profound hypoglycemia, We have discovered that islet cells, isolated from the pancreas of a PHHI patient, proliferate in culture while maintaining a beta cell-like phenotype, The PHHI-derived cell line (NES2Y) exhibits insulin secretory characteristics typical of islet cells derived from these patients, i,e, they have no K-ATP channel activity and as a consequence secrete insulin at constitutively high levels in the absence of glucose, In addition, they exhibit impaired expression of the homeodomain transcription factor PDX1, which is a key component of the signaling pathway linking nutrient metabolism to the regulation of insulin gene expression. To repair these defects NES2Y cells were triple-transfected with cDNAs encoding the two components of the K-ATP channel (SUR1 and Kir6.2) and PDX1, One selected clonal cell line (NISK9) had normal K-ATP channel activity, and as a result of changes in intracellular Ca2+ homeostasis ([Ca2+](i)) secreted insulin within the physiological range of glucose concentrations. This approach to engineering PHHI-derived islet cells may be of use in gene therapy for PHHI and in cell engineering techniques for administering insulin for the treatment of diabetes mellitus.