Transfection of NFκB-decoy oligodeoxynucleotides using efficient ultrasound-mediated gene transfer into donor kidneys prolonged survival of rat renal allografts

Abstract

Nuclear factor kB (NFkB) plays a pivotal role in the coordinated transactivation of a series of genes of cytokines and adhesion molecules that are highly involved in the onset of acute rejection in organ transplantation. We previously developed decoy cis-elements oligo deoxyribonucleic acid against NFkB (NFkB-decoy) that effectively inhibited the activation of major inflammatory mediators in vitro and in vivo. Accordingly, we hypothesized that transfection of NFkB-decoy into the donor kidney would prevent acute rejection and prolong graft survival, and thus provide effective therapy for renal acute rejection. To transfect NFkB-decoy, we employed a novel approach using ultrasound exposure with an echocardiographic contrast agent, Optison, and clearly demonstrated successful transfection of NFkB-decoy into renal tissue. The therapeutic effect of NFkB-decoy on renal allografts was then evaluated in a rat renal allograft model (Wistar-Lewis). In the control group, graft function significantly deteriorated with marked destruction of renal tissue, accompanied by increased production of major inflammatory mediators, and all animals died of renal failure by 9 days. In contrast, graft function (serum creatinine on day 2, NFkB-treated: 0.9770.16 versus control: 1.8470.23 mg/dl, Po0.01) and histological structure were well preserved with significantly decreased expression of NFkB-regulated cytokines and adhesion molecules, including IL-1, iNOS, MCP-1, TNF-a, and ICAM-1, in allografts transfected with NFkB-decoy. As a result, animal survival was significantly prolonged in this group as compared to controls (14.275.2 versus 7.171.2 days, Po0.01). Thus, we established a novel ultrasound-Optison-mediated gene transfection approach and demonstrated the significant prolongation of graft survival by the successful transfection of NFkB-decoy into the donor kidney in a rat renal allograft model. Gene Therapy (2003) 10, 415-425.

Gene therapy progress and prospects: Ultrasound for gene transfer

“…Most in vivo studies have concentrated on organs and tissues that are readily imaged by diagnostic ultrasound machines, including skeletal muscle, heart and kidneys. 35,36,[48][49][50][51][52][53][54][55][56][57][58][59][60][61][62] Table 1 summarizes the results of some recent studies in these tissues using reporter genes, 35,36,[48][49][50][51][52][53][54]58,60,61 and broadly similar data have begun to emerge from studies in the vascular, 4,63,64 neurology 65-67 and oncology 18,37,68 fields, the pancreas 69 and in hematology. 70 On first inspection the relatively modest enhancements in plasmid-mediated transfection after USE in vivo may appear disappointing compared with the results obtained in vitro.…”

Key factors that affect sonoporation efficiency in in vitro settings: The importance of standing wave in sonoporation

“…Although there are many reports on sonoporation [21][22][23][24][25][26][27][28][29][30][31][32][33][34][35][36][37][38], each experimental setting differs from one another and the reported sonoporation efficiency, which is one of the most significant information in sonoporation, varies. In order to understand the effect of the experimental settings themselves on sonoporation experiments, we first investigated the impact of the conditions of the cultured cells on sonoporation efficiency.…”

“…(Current address: Department of Neurosurgery, Osaka University Medical School, 2-2 Yamadaoka, Suita, Oaska, 565-0871, Japan. Phone: +81-6-6879-3652: FAX: +81-6-6879-3659; e-mail : m-kinoshita@nsurg.med.osaka-u.ac.jp) hyperthermia, such as coagulation of tissues [1][2][3][4][5][6][7][8][9][10][11][12], mechnical tissue destruction [13,14], disruption of tight junctions of the vasculature for enhanced local drug delivery [15][16][17][18][19][20], and sonoporation, a transient enhanced permeability of the cell membrane caused by the combination of ultrasound and microbubbles [21][22][23][24][25][26][27][28][29][30][31][32][33][34][35][36][37][38]. As all of these bio-effects can be created non-invasively in vivo, ultrasound is gathering a great attention as a next generation surgical or drug delivery system.…”

Pharmacology in Health Foods: Improvement of Vascular Endothelial Function by French Maritime Pine Bark Extract (Flavangenol)

“…It has been well known that the transcriptional factor nuclear factor-kappa B (NF-κB) is mainly responsible for the regulation of these gene expressions (40). We and others have demonstrated that the suppression of NF-κB could become a novel therapeutic strategy for ischemia/ reperfusion-induced tissue damage (41,42). There is accumulating evidence that PBE has an inhibitory effect on NF-κB activation in macrophages and vascular endothelial cells (43,44).…”