Reversible expansion of pancreatic islet progenitors derived from human induced pluripotent stem cells
Reversible expansion of pancreatic islet progenitors derived from human induced pluripotent stem cells
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DOI：
10.1111/gtc.12759
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发表时间：
2020-05-01
期刊：
影响因子：
2.1
通讯作者：
Miyajima, Atsushi
中科院分区：
文献类型：
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作者：
Tanaka, Anna;Watanabe, Ami;Miyajima, Atsushi
胰岛移植是治疗重症1型糖尿病的有效方法。由于供体短缺是这种疗法的主要问题，因此已经尝试从人多能干细胞（hPSC）产生大量胰岛。然而，由于hPSC向胰岛的分化需要使用各种昂贵的细胞因子的多个和冗长的过程，因此该过程是可变的、低效率的和昂贵的。因此，如果能够扩增胰岛祖细胞将是有益的。来源于hPSC的表达神经生成素3（NGN 3）的胰腺内分泌祖细胞（EP）表现出分化成胰岛的能力，同时其细胞周期被阻滞。通过使用慢病毒载体，我们将几个生长促进基因引入表达NGN 3的EP细胞中。我们发现SV 40 LT的表达诱导了EP细胞的增殖，但降低了内分泌谱系承诺因子NGN 3、NEUROD 1和NKX2.2的表达，导致胰岛分化的抑制。通过使用Cre-loxP系统，我们在扩增后去除了SV 40 LT，导致内分泌谱系定型基因的重新表达和分化成功能性胰岛。因此，我们的研究结果将为通过从hPSC扩增EP细胞来产生大量功能性胰岛铺平道路。
Transplantation of pancreatic islets is an effective therapy for severe type 1 diabetes. As donor shortage is a major problem for this therapy, attempts have been made to produce a large number of pancreatic islets from human pluripotent stem cells (hPSCs). However, as the differentiation of hPSCs to pancreatic islets requires multiple and lengthy processes using various expensive cytokines, the process is variable, low efficiency and costly. Therefore, it would be beneficial if islet progenitors could be expanded. Neurogenin3 (NGN3)-expressing pancreatic endocrine progenitor (EP) cells derived from hPSCs exhibited the ability to differentiate into pancreatic islets while their cell cycle was arrested. By using a lentivirus vector, we introduced several growth-promoting genes into NGN3-expressing EP cells. We found that SV40LT expression induced proliferation of the EP cells but reduced the expression of endocrine lineage-commitment factors, NGN3, NEUROD1 and NKX2.2, resulting in the suppression of islet differentiation. By using the Cre-loxP system, we removed SV40LT after the expansion, leading to re-expression of endocrine-lineage commitment genes and differentiation into functional pancreatic islets. Thus, our findings will pave a way to generate a large quantity of functional pancreatic islets through the expansion of EP cells from hPSCs.