Enhanced protection of Ins-1 β cells from apoptosis under hypoxia by delivery of DNA encoding secretion signal peptide-linked exendin-4 Exendin-4

Hyun Ah Kim, Suyeon Lee, Jeong Hyun Park, Sanghyun Lee, Byung Wan Lee, Sung Hee Ihm, Tae Il Kim, Sung Wan Kim, Kyung Soo Ko, Minhyung Lee

Research output: Contribution to journalArticlepeer-review

18 Citations (Scopus)

Abstract

In this study, we developed an expression system of exendin-4, a glucagon-like peptide (GLP-1) analog, using a secretion signal peptide (SP) to facilitate exendin-4 secretion. For delivery of the exendin-4 expression system, high-molecular-weight polyethylenimine (25 kDa, PEI25k), low-molecular-weight polyethylenimine (2 kDa, PEI2k), and polyamidoamine (PAMAM) dendrimers were evaluated as gene carriers to Ins-1 β cells. As a result, PEI25k showed the highest transfection efficiency. For the construction of the exendin-4 expression vector, DNA coding the SP sequence was inserted upstream of the exendin-4 cDNA, resulting in the construction of pβ-SP-Ex-4. Transfection assay showed that the secretion level of exendin-4 increased in the pβ-SP-Ex-4 transfected cells, compared with the pβ-Ex-4 transfected cells. To identify the β-cell protection effect of pβ-SP-Ex-4 delivery, the Ins-1 β cells were transfected with pβ-SP-Ex-4 or pβ-Ex-4 and incubated under normoxia or hypoxia. An MTT assay showed that the pβ-SP-Ex-4 transfected cells had higher β-cell viability than the pβ-Ex-4 transfected cells under hypoxia. In addition, the pβ-SP-Ex-4 transfected cells exhibited lower caspase-3 activity than the pβ-Ex-4 transfected cells. Therefore, PEI25k/pβ-SP-Ex-4 complex may be useful to protect isolated β cells from apoptosis during transplantation.

Original languageEnglish
Pages (from-to)242-248
Number of pages7
JournalJournal of drug targeting
Volume17
Issue number3
DOIs
Publication statusPublished - 2009 Apr

Bibliographical note

Funding Information:
This work was supported by the Research Funds from Korean Diabetes Association, the Innovative Research Institute for Cell Therapy, Republic of Korea (A062260), and NIH grant (DK077703).

All Science Journal Classification (ASJC) codes

  • Pharmaceutical Science

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