| 학회 | 한국고분자학회 |
| 학술대회 | 2005년 가을 (10/13 ~ 10/14, 제주 ICC) |
| 권호 | 30권 2호 |
| 발표분야 | 고분자를 이용한 유전자 전달 |
| 제목 | Polymer mediated gene delivery for type 2 diabetes gene therapy |
| 초록 | Glucagon-like peptide 1 (GLP-1) is a insulinotropic protein, which increases the insulin secretion from islet β-cells at a high glucose concentration. It was reported that the continuous infusion of GLP-1 normalized the blood glucose level in type 2 diabetes animal model1. However, the extremely short half-life of GLP-1 has limited its application2 and prompted the gene therapy with the GLP-1 expression plasmid3. pβ-GLP-1, a GLP-1 expression plasmid, was transfected to cells using polyethylenimine (PEI) as a gene carrier. The in vitro results showed a dose dependent expression of GLP-1. Co-culture assay of the GLP-1 plasmid transfected cells with isolated rat islet cells demonstrated that GLP-1 increased insulin secretion, compared to controls during a hyperglycemic challenge. A single injection of PEI/pβ-GLP-1 complex into ZDF rats resulted in increasing insulin secretion and decreasing blood glucose level that was maintained for 2 weeks. However, the glucose level was not completely normalized, suggesting that the expression level of GLP-1 was not enough to normalize the blood glucose level. More efficient GLP-1 expression systems were developed using two step transcription amplification (TSTA) or nuclear factor-κb (NF-κb) mediated nuclear transport. To apply the TSTA system to the GLP-1 expression plasmid, pβ-Gal4-p65 and pUAS-GLP-1 were constructed and co-transfected to 293 cells using PEI as a gene carrier. The pβ-Gal4-p65/pUAS-GLP-1 showed higher mRNA expression than pβ-GLP-1 in RT-PCR. In addition, the GLP-1 protein level by pβ-Gal4-p65/pUAS-GLP-1 was 4 times higher than pβ-GLP-1. The highest transgene expression was obtained at a 2:1 pUAS-GLP-1:pβ-Gal4-p65 ratio. The ratio between the two plasmids had significant effect of the trasngene expression. In the second strategy, NF-κb binding sites were introduced into the GLP-1 expression plasmid. The in vitro results showed higher expression of GLP-1 compared to the control plasmid. A single systemic administration of PEI/pSi-GLP-1-NF-κb complex into DIO mice resulted in increasing insulin secretion and decresing blood glucose levels, compared to the control GLP-1 plasmid without NF-κb binding sites. This GLP-1 gene delivery system may provide an effective treatment modality for type 2 diabetes. 참고문헌 1. M. A. Nauck, A. Sauerwald, R. Ritzel, J. J. Holst, and W. Schmiegel, Diabetes Care, 21, 1925 (1998). 2. J. J. Holst, Gastroenterology, 107, 1848 (1994). 3. S. Oh, M. Lee, K.S. Ko, S. Choi and S.W. Kim, Mol. Ther., 7, 478 (2003). |
| 저자 | 이민형1, 오승준2, 고경수3, 최선아4, 안철희5, 김성완4 |
| 소속 | 1한양대, 2경희대, 3인제대, 4유타대, 5서울대 |
| 키워드 | glucagon like peptide-1; polyethylenimine; gene therapy; type 2 diabetes |
