摘要
将尸体捐赠者的胰岛移植至病人体内可成功地使血糖正常化,这一概念证明1型糖尿病是可治愈的。尽管如此,但要说明的是,这种细胞疗法广泛使用的主要问题为移植后的胰岛细胞缺乏活性及功能不足。基因治疗可在移植前有针对性的提高活性和性能,并提供一个可行的方法来规避上述问题,以维持体内β细胞群功能持久。然而,将核酸安全有效地转移至胰岛仍然是一项具有挑战性的任务。在这里,我们描述一个简单易用的慢病毒载体介导方案,可介导大约80%的老鼠和人胰岛细胞,同时保留胰岛结构,代谢功能和葡萄糖依赖性刺激胰岛素的分泌。我们的方案将促进完全确定整个胰岛细胞异种移植过程中的潜在治疗靶点及基因表达调控。
关键词: 糖尿病,转基因,传染,慢病毒,胰岛,介导
Current Gene Therapy
Title:A Simple High Efficiency Intra-Islet Transduction Protocol Using Lentiviral Vectors
Volume: 15 Issue: 4
Author(s): Carmen Maria Jimenez-Moreno, Irene de Gracia Herrera-Gomez, Livia Lopez-Noriega, Petra Isabel Lorenzo, Nadia Cobo-Vuilleumier, Esther Fuente-Martin, Jose Manuel Mellado-Gil, Geraldine Parnaud, Domenico Bosco and Benoit Raymond Gauthier and Alejandro Martin-Montalvo
Affiliation:
关键词: 糖尿病,转基因,传染,慢病毒,胰岛,介导
摘要: Successful normalization of blood glucose in patients transplanted with pancreatic islets isolated from cadaveric donors established the proof-of-concept that Type 1 Diabetes Mellitus is a curable disease. Nonetheless, major caveats to the widespread use of this cell therapy approach have been the shortage of islets combined with the low viability and functional rates subsequent to transplantation. Gene therapy targeted to enhance survival and performance prior to transplantation could offer a feasible approach to circumvent these issues and sustain a durable functional β-cell mass in vivo. However, efficient and safe delivery of nucleic acids to intact islet remains a challenging task. Here we describe a simple and easy-to-use lentiviral transduction protocol that allows the transduction of approximately 80 % of mouse and human islet cells while preserving islet architecture, metabolic function and glucose-dependent stimulation of insulin secretion. Our protocol will facilitate to fully determine the potential of gene expression modulation of therapeutically promising targets in entire pancreatic islets for xenotransplantation purposes.
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Carmen Maria Jimenez-Moreno, Irene de Gracia Herrera-Gomez, Livia Lopez-Noriega, Petra Isabel Lorenzo , Nadia Cobo-Vuilleumier, Esther Fuente-Martin, Jose Manuel Mellado-Gil, Geraldine Parnaud, Domenico Bosco and Benoit Raymond Gauthier and Alejandro Martin-Montalvo , A Simple High Efficiency Intra-Islet Transduction Protocol Using Lentiviral Vectors, Current Gene Therapy 2015; 15 (4) . https://dx.doi.org/10.2174/1566523215666150630121557
DOI https://dx.doi.org/10.2174/1566523215666150630121557 |
Print ISSN 1566-5232 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5631 |
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