在用人类胰岛素原基因（p3MTChins）电穿孔的原代小鼠肝细胞移植后，非肥胖糖尿病小鼠和链脲佐菌素诱导的糖尿病小鼠未能实现正常的代谢反应。

Failure to achieve normal metabolic response in non-obese diabetic mice and streptozotocin-induced diabetic mice after transplantation of primary murine hepatocytes electroporated with the human proinsulin gene (p3MTChins).

作者信息

Lee R H, Roll G, Nguyen V, Willenbring H, Tang Q, Kang S-M, Stock P G

机构信息

Department of Surgery, University of California-San Francisco, San Francisco, California, USA.

出版信息

Transplant Proc. 2014 Jul-Aug;46(6):2002-6. doi: 10.1016/j.transproceed.2014.05.068.

DOI:10.1016/j.transproceed.2014.05.068

PMID:25131094

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5262438/

Abstract

BACKGROUND

A recent study by Chen et al described a therapy for diabetes that involved electroporation of primary hepatocytes with human proinsulin cDNA, p3MTChins. Intrahepatic transplantation of treated hepatocytes into streptozotocin (STZ) murine and porcine models led to euglycemia, weight maintenance, and normal insulin production. We tested the repeatability of their basic experiments and transplantation technique and expanded the study to include an autoimmune model.

METHODS

Hepatocytes were isolated from B6 mice, electroporated with p3MTChins, and glucose-challenged or were injected into hepatic or spleen parenchyma of STZ-diabetic B6 and non-obese diabetic mice. Outcomes included survival, serum glucose levels, insulin, and c-peptide release. Untransfected primary hepatocytes and mice transplanted with these cells served as controls.

RESULTS

p3MTChins-hepatocytes secreted insulin during glucose challenge, but glucose levels did not change with increasing glucose concentrations. Direct hepatic injection led to high mortality rates. Mice that underwent intrasplenic injection survived for >50 days (control = 4 days) and had a mild but stable improvement in hyperglycemia. C-peptide in both mouse models was detectable but eventually declined to baseline in the non-obese diabetic mice.

CONCLUSIONS

Hepatocytes can be transfected with p3MTChins to produce human insulin but may lack the proper glucose-sensing or complex storage and secretion capabilities that allow for a finely tuned dynamic insulin response. Treatment is subtherapeutic, and p3MTChins-hepatocyte function may not endure in an autoimmune model. Without successful preliminary findings, cell therapy involving electroporation of p3MTChins does not appear to be practical as a therapy for diabetes and may not be a strategy to pursue at this time.

摘要

背景

陈等人最近的一项研究描述了一种糖尿病治疗方法，该方法涉及用人胰岛素原cDNA（p3MTChins）对原代肝细胞进行电穿孔。将经处理的肝细胞肝内移植到链脲佐菌素（STZ）诱导的小鼠和猪模型中，可导致血糖正常、体重维持以及胰岛素正常分泌。我们测试了他们基础实验和移植技术的可重复性，并将研究扩展至包括一个自身免疫模型。

方法

从B6小鼠中分离肝细胞，用p3MTChins进行电穿孔，然后进行葡萄糖激发试验，或将其注射到STZ诱导的糖尿病B6小鼠和非肥胖糖尿病小鼠的肝实质或脾实质中。观察指标包括生存率、血清葡萄糖水平、胰岛素和C肽释放。未转染的原代肝细胞以及移植了这些细胞的小鼠作为对照。

结果

p3MTChins转染的肝细胞在葡萄糖激发试验期间分泌胰岛素，但血糖水平并未随葡萄糖浓度升高而变化。直接肝内注射导致高死亡率。接受脾内注射的小鼠存活超过50天（对照组=4天），高血糖有轻度但稳定的改善。两种小鼠模型中的C肽均可检测到，但在非肥胖糖尿病小鼠中最终降至基线水平。

结论

肝细胞可用p3MTChins转染以产生人胰岛素，但可能缺乏适当的葡萄糖感应或复杂的储存及分泌能力，从而无法实现精细调节的动态胰岛素反应。该治疗效果欠佳，且p3MTChins转染的肝细胞功能在自身免疫模型中可能无法持久。由于没有成功的初步结果，涉及p3MTChins电穿孔的细胞疗法作为糖尿病治疗方法似乎并不实用，目前可能不是一个值得 pursue 的策略。（注：pursue在这里根据语境推测可能是“继续、推行”等意思，但原文此处似乎表述不太完整准确）