Lentivirus-mediated Bcl-2 expression in βTC-tet cells improves resistance to hypoxia and cytokine-induced apoptosis while preserving in vitro and in vivo control of insulin secretion
Lentivirus-mediated Bcl-2 expression in βTC-tet cells improves resistance to hypoxia and cytokine-induced apoptosis while preserving in vitro and in vivo control of insulin secretion
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DOI：
10.1038/sj.gt.3300922
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发表时间：
1999-06-01
期刊：
影响因子：
5.1
通讯作者：
Thorens, B
中科院分区：
文献类型：
--
作者：
Dupraz, P;Rinsch, C;Thorens, B
β TC-tet细胞是条件性永生化的胰腺β细胞，当移植到同基因链脲佐菌素糖尿病小鼠中时，其可以长期纠正高血糖症。使用这些细胞来控制人类I型糖尿病将需要将它们封装并移植到相对缺氧和细胞因子可能威胁其存活的非天然部位。在这项研究中，我们使用新的慢病毒载体用抗凋亡基因Bcl-2对β TC-tet细胞进行基因工程改造，并显示其保护该细胞系免受由缺氧、星形孢菌素和细胞因子（IL-1 β、IFN-γ和TNF-α）的混合物诱导的凋亡。我们进一步证明，Bcl-2的表达允许生长在更高的细胞密度和更短的倍增时间。Bcl-2的表达，但是，没有干扰的内在机制的生长停滞存在于β TC-tet细胞或其正常的葡萄糖剂量依赖性胰岛素分泌活性。此外，表达Bcl-2的β TC-tet细胞在轻度缺氧条件下仍保持其分泌胰岛素的能力。最后，将这些细胞移植到链脲佐菌素糖尿病C3 H小鼠的肾包膜下，纠正了几个月的高血糖。这些结果表明，鼠β TC-tet细胞系可以进行遗传修饰，以提高其对不同应激诱导的细胞凋亡的抗性，同时保持其正常的生理功能。这些修饰的细胞代表了I型糖尿病细胞移植治疗的改进来源。
beta TC-tet cells are conditionally immortalized pancreatic beta cells which can confer long-term correction of hyperglycemia mia when transplanted in syngeneic streptozocin diabetic mice. The use of these cells for control of type I diabetes in humans will require their encapsulation and transplantation in non-native sites where relative hypoxia and cytokines may threaten their survival. in this study we genetically engineered beta TC-tet cells with the anti-apoptotic gene Bcl-2 using new lentiviral vectors and showed that it protected this cell line against apoptosis induced by hypoxia, staurosporine and a mixture of cytokines (IL-1 beta, IFN-gamma and TNF-alpha). We further demonstrated that Bcl-2 expression permitted growth at higher cell density and with shorter doubling time. Expression of Bcl-2, however, did not interfere either with the intrinsic mechanism of growth arrest present in the beta TC-tet cells or with their normal glucose dose-dependent insulin secretory activity. Furthermore, Bcl-2 expressing beta TC-tet cells retained their capacity to secrete insulin under mild hypoxia. Finally, transplantation of these cells under the kidney capsule of streptozocin diabetic C3H mice corrected hyperglycemia for several months. These results demonstrate that the murine beta TC-tet cell line can be genetically modified to improve its resistance against different stress-induced apoptosis while preserving its normal physiological function. These modified cells represent an improved source for cell transplantation therapy of type I diabetes.