Regulated insulin delivery from human epidermal cells reverses hyperglycemia.
Regulated insulin delivery from human epidermal cells reverses hyperglycemia.
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DOI：
10.1038/mt.2008.79
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发表时间：
2008-06
期刊：
影响因子：
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通讯作者：
Jun Tian;P. Lei;S. G. Laychock;S. Andreadis
中科院分区：
文献类型：
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作者：
Jun Tian;P. Lei;S. G. Laychock;S. Andreadis
正在寻找替代胰岛素疗法，为糖尿病患者提供正常血糖控制。表皮是一种自我更新的组织，易于获取，可以提供大量自体细胞，可用于产生分泌胰岛素的皮肤替代品。慢病毒载体已被改造，以产生弗林蛋白酶可切割的胰岛素原和FK 506结合蛋白的自身二聚化突变体之间的融合蛋白，从而在角质形成细胞中产生生物活性胰岛素;这种胰岛素作为对外源性施用小有机分子雷帕霉素的响应而释放。工程化的角质形成细胞保持正常的形态，并以类似于慢病毒处理的对照细胞的方式生长。培养物和分层生物工程表皮中的表皮角质形成细胞在加入雷帕霉素后30分钟内释放胰岛素，并且在去除诱导剂后2-3小时内分泌减缓或停止。当将细胞植入用链脲佐菌素（STZ）致糖尿病的无胸腺小鼠中时，在加入雷帕霉素后1小时内在血浆中检测到胰岛素。同时，即使在具有严重高血糖症的糖尿病动物中，血清葡萄糖也降至正常水平。重复雷帕霉素给药产生了类似的结果。这些实验提供了概念验证，即以可调节的方式从皮肤释放的胰岛素可以逆转高血糖症。
Alternative insulin therapies are being sought that will provide euglycemic control for people with diabetes mellitus. The epidermis is a self-renewing tissue that is easily accessible and can provide large numbers of autologous cells that can be used for generating insulin-secreting skin substitutes. Lentiviral vectors have been engineered to produce a fusion protein between the furin-cleavable proinsulin and the self-dimerization mutant of FK506-binding protein to yield bioactive insulin in keratinocytes; this insulin is released as a response to exogenous administration of a small organic molecule, rapamycin. The engineered keratinocytes retained normal morphology and grew in a manner similar to lentiviral-treated control cells. Epidermal keratinocytes in culture and in stratified bioengineered epidermis released insulin within 30 minutes after addition of rapamycin, and secretion slowed or stopped within 2–3 hours after removal of the inducing agent. When the cells were implanted into athymic mice that had been rendered diabetic with streptozotocin (STZ), insulin was detected in the plasma within 1 hour after addition of rapamycin. Concomitantly, serum glucose decreased to normal levels even in diabetic animals with severe hyperglycemia. Repeated rapamycin administration yielded similar results. These experiments provide proof-of-concept that insulin released from the skin in a regulatable manner can reverse hyperglycemia.