Transduction of human and mouse pancreatic islet cells using a bicistronic recombinant adeno-associated viral vector

被引:28
作者
Kapturczak, M
Zolotukhin, S
Cross, J
Pileggi, A
Molano, RD
Jorgensen, M
Byrne, B
Flotte, TR
Ellis, T
Inverardi, L
Ricordi, C
Nick, H
Atkinson, M
Agarwal, A [1 ]
机构
[1] Univ Florida, Dept Med, Gainesville, FL 32610 USA
[2] Univ Florida, Powell Gene Therapy Ctr, Gainesville, FL 32610 USA
[3] Univ Florida, Dept Pathol, Gainesville, FL 32610 USA
[4] Univ Florida, Dept Pediat, Gainesville, FL 32610 USA
[5] Univ Florida, Dept Neurosci, Gainesville, FL 32610 USA
[6] Univ Miami, Diabet Res Inst, Miami, FL 33136 USA
关键词
islet transplantation; gene therapy; diabetes; adeno-associated virus; bicistronic vector; transplant rejection; autoimmunity;
D O I
10.1006/mthe.2002.0522
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Recent reports indicate successful transduction of pancreatic islets using recombinant adeno-associated viral (rAAV) vectors. This advance offers new possibilities in rendering islets resistant to rejection and recurrence of autoimmune destruction in the setting of islet transplantation as treatment of type 1 diabetes. Most gene delivery approaches using islets have thus far involved transduction with a single gene. However, the concomitant delivery of more than one gene encoding cytoprotective and/or immunoregulatory molecules may offer superior clinical utility. Here, we have generated a bicistronic rAAV (serotype 2) vector incorporating a viral internal ribosome entry site (IRES), derived from polio virus type 1, to allow for translation of two coupled cDNAs from a single mRNA transcript. Our study demonstrates the ability of this vector to produce significant expression of two reporter proteins in human and mouse islets in vitro. This expression did not interfere with beta-cell function. Transduction was maintained in vivo following transplantation of mouse islets. These data are the first report of efficient islet cell transduction with two genes using a single bicistronic rAAV vector and have direct implications for strategies aimed at enhancing islet transplant survival.
引用
收藏
页码:154 / 160
页数:7
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