Isolated Limb Perfusion for Local Gene Delivery (original) (raw)
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Intra-arterial adenoviral mediated tumor transfection in a novel model of cancer gene therapy
Molecular cancer, 2006
The aim of the present study was to develop and characterize a novel in vivo cancer gene therapy model in which intra-arterial adenoviral gene delivery can be characterized. In this model, the rat cremaster muscle serves as the site for tumor growth and provides convenient and isolated access to the tumor parenchyma with discrete control of arterial and venous access for delivery of agents. Utilizing adenovirus encoding the green fluorescent protein we demonstrated broad tumor transfection. We also observed a dose dependent increment in luciferase activity at the tumor site using an adenovirus encoding the luciferase reporter gene. Finally, we tested the intra-arterial adenovirus dwelling time required to achieve optimal tumor transfection and observed a minimum time of 30 minutes. We conclude that adenovirus mediated tumor transfection grown in the cremaster muscle of athymic nude rats via an intra-arterial route could be achieved. This model allows definition of the variables that...
Gene Therapy, 1997
Targeted gene delivery is essential for gene therapy involv-Perfusion with chelating agents (1 mM EGTA, or 2 mM ing in vivo gene transfer. In the present study we analyzed EDTA) prior to the administration of the vector increased the efficiency and tissue-specificity of gene transfer into the the efficiency to at least 40%. Similar efficiencies were liver with recombinant adenoviruses. Adenovirus vectors obtained in experiments with liver lobes of Rhesus monkcarrying the E. coli lacZ gene (Ad.RSV.-gal) and the firefly eys. In vivo administration of AdCMV-luc via ILP resulted luciferase gene (AdCMV-luc) as reporters were adminis-in a significantly more efficient (P = 0.028) and also more tered to the liver of adult Wistar rats, either via infusion into reproducible gene transfer when compared to IPI. Although the portal vein (intraportal infusion; IPI) or via perfusion of detectable in both groups, extrahepatic luciferase the vascularly isolated liver (isolated liver perfusion; ILP). expression was considerably reduced in the ILP group. Our Ex vivo liver perfusion experiments with Ad.RSV.-gal data demonstrate that IPL can be used for efficient and were used to optimize the conditions for hepatic gene reproducible liver-specific gene delivery. Therefore, we transfer. Ex vivo perfusion of rat livers with 2 × 10 9 plaque think that the perfusion of vascularly isolated organs is useforming units (p.f.u.) Ad.RSV.-gal was sufficient to ful as a modality for the tissue-specific administration of infect about 20% of the liver parenchymal cells. recombinant adenovirus vectors.
Faseb Journal, 2000
Expression of transgene other than in the target tissue may cause side effects and safety problems in gene therapy. We analyzed biodistribution of transgene expression after intravascular and periadventitial gene delivery methods using the first generation nuclear-targeted lacZ adenovirus. RT-PCR and X-Gal stainings were used to study transgene expression 14 days after the gene transfer. After intravascular catheter-mediated gene transfer to rabbit aorta mimicking angioplasty procedure, the target vessel showed 1.1% ؎ 0.5 gene transfer efficiency. Other tissues showed varying lacZ gene expression indicating a systemic leakage of the vector with the highest transfection efficiency in hepatocytes (0.7% ؎ 0.5). X-Gal staining of blood cells 24 h after the intravascular gene transfer indicated that a significant portion (1.8% ؎ 0.8) of circulating monocytes was transfected. X-Gal-positive cells were also found in testis. After periadventitial gene transfer using a closed silicon capsule placed around the artery, 0.1% ؎ 0.1 lacZ-positive cells were detected in the artery wall. Positive cells were also found in the liver and testis (<0.01%), indicating that the virus escapes even from the periadventitial space, although less extensively than during the intravascular application. We conclude that catheter-mediated intravascular and, to a lesser extent, periadventitial gene transfer lead to leakage of adenovirus to systemic circulation, followed by expression of the transgene in several tissues. Possible consequences of the ectopic expression of the transgene should be evaluated in gene therapy trials even if local gene delivery methods are used.-Hiltunen, M. O., Turunen, M. P., Turunen, A.-M., Rissanen, T. T., Laitinen, M., Kosma, V.-M., Ylä-Herttuala, S. Biodistribution of adenoviral vector to nontarget tissues after local in vivo gene transfer to arterial wall using intravascular and periadventitial gene delivery methods. FASEB J. 14, 2230 -2236 (2000)
Gene Therapy, 2008
Transfer of healthy autologous tissue as a microvascular free flap facilitates reconstruction during ablative cancer surgery. In addition to filling surgical defects, free flaps might concentrate viral vectors at the tumour bed and mediate local therapeutic effects. We evaluated the magnitude, topography and duration of luciferase gene expression after plasmid and adenoviral delivery in rat superficial inferior epigastric (SIE) flaps. For plasmid delivery, luciferase expression was significantly increased by all transduction routes (topical, intraflap injection, intravascular) (Po0.01) at day 1, but not at day 7. The spread of luciferase expression was significantly different between the 4 groups at 1 day (P ¼ 0.026) and was greatest for flaps transduced by intravascular injection. For adenoviral transduction, total radiance was significantly different between the transduced groups at 1, 14 and 28 days (Po0.05 for all comparisons). The highest levels of radiance were seen in the intravascular group. There was a statistically significant difference in the spread of light emission between the 3 groups at 1 (P ¼ 0.009) and 14 (P ¼ 0.013) days, but this was no longer evident at 28 days. Intravascular adenoviral delivery yields high-level, diffuse and durable gene expression in rat SIE flaps and is suitable for examination in therapeutic models.
Cancer Gene Therapy, 2003
The immune response is modulated by genetic adjuvants using plasmid vectors expressing cytokines. Skeletal muscle can express a foreign gene intramuscularly administered via a needle injection, and the potential of muscle as a target tissue for somatic gene therapy in treating cancer has been explored. In the present study, we investigated the efficacy of particle-mediated intramuscular transfection modified with a local anesthetic agent, bupivacaine, on luciferase and green fluorescent protein. The results indicate that these proteins are more efficiently expressed and persist longer in muscle modified in this way compared with the needleinjection method. Using an established rat sarcoma model, particle-mediated intramuscular gene-gun therapy with a combination of IL-12 and IL-18 cDNA was conducted. Growth of the distant sarcoma was significantly inhibited by particle-mediated intramuscular combination gene therapy, and the survival rate was also improved. Furthermore, the combination gene-gun therapy maintained significant levels of interferon-g and induced a high activity of tumor-specific cytotoxic T lymphocytes. These results suggest that the sustained local delivery of IL-12 and IL-18 cDNA using intramuscular gene-gun therapy modified with bupivacaine can induce long-term antitumor immunity, and can provide the great advantage of inhibiting the disseminated tumor.
Locoregional delivery of adenoviral vectors
Journal of Nuclear …, 2006
The overall median survival of patients with a malignant glioma is ,1 y. Because malignant gliomas rarely metastasize outside the skull, locoregional treatment strategies, such as gene therapy, are under investigation. Recently, convection-enhanced delivery (CED) has been presented as a method to improve delivery of large molecules. The goal of this study was to evaluate whether CED improves intratumoral delivery of adenoviral vectors and compare it with single injection (SI) and multiple injection (4•, MI). Methods: A replication-deficient adenoviral vector encoding the herpes simplex virus thymidine kinase (HSV-tk) and the human somatostatin receptor subtype 2 (sst 2) was administered into nude mice bearing subcutaneous U87 xenografts. Tumors were injected with 1.5 • 10 9 plaque-forming units of Ad5.tk.sstr by CED, SI, or MI. Three days later, [ 99m Tc-N 4 0-1 ,Asp 0 ,Tyr 3 ] octreotate (99m Tc-Demotate 2) was injected intravenously to monitor the virus-induced sst 2 expression. g-Camera imaging was performed for in vivo imaging, and the tumor uptake of 99m Tc-Demotate 2 was determined by g-counter. Furthermore, the tumor was sectioned and ex vivo autoradiography was performed. After decay of radioactivity, adjacent sections were submitted to in vitro autoradiography with 125 I-DOTA-Tyr 3-octreotate, which was used to calculate the transduced areas. Results: Transfected xenograft tissues showed high sst 2 expression and were clearly visualized with a g-camera. Accumulation of radioactivity was 2-fold higher in the tumors that were injected with MI compared with CED and SI (P 5 0.01). CED and SI resulted in equal uptake of radioactivity in the tumors. The measured areas of transduction in ex vivo and in vitro autoradiographs showed a high concordance (r 2 5 0.89, P , 0.0001). The maximum area of transfection was significantly larger after MI than after CED (P , 0.05) or SI (P 5 0.05). Also, the measured volume of distribution was twice as high after administration of Ad5.tk.sstr by MI (56.6 mm 3) compared with SI (25.3 mm 3) or CED (26.4 mm 3). Conclusion: CED does not increase adenoviral vector distribution in a glioma xenograft model compared with SI. Therefore, in the clinic MI is probably the most effective delivery method for the large adenoviral particle (70 nm) in malignant gliomas.
Adenovirus-mediated delivery of antiangiogenic genes as an antitumor approach
Cancer Gene Therapy, 2000
Based on the observation that the growth of solid tumors is dependent on the formation of new blood vessels, therapeutic strategies aimed at inhibiting angiogenesis have been proposed. A number of proteins with angiostatic activity have been described, but their development as therapeutic agents has been hampered by difficulties in their production and their poor pharmacokinetics. These limitations may be resolved using a gene therapy approach whereby the genes are delivered and expressed in vivo. Here we compared adenoviral delivery of endostatin, proliferin -related protein ( PRP ) , and interferon -inducible protein 10 ( IP10 ) genes. Recombinant adenoviruses carrying the three angiostatic genes express biologically active gene products as determined in vitro in endothelial cell proliferation and migration assays, and in vivo by inhibition of neoangiogenesis in rat chambers. Eradication of established tumors in vivo, in the murine B16F10 melanoma model in immunocompetent mice, was not achieved by intratumoral injection of the different vectors. However, the combination of intravenous plus intratumoral injections allowed rejection of tumors. Ad -PRP or Ad -IP10 were significantly more efficient than Ad -endostatin, leading to complete tumor rejection and prolonged survival in a high proportion of treated animals. These data support the use of in vivo gene delivery approaches to produce highcirculating and local levels of antiangiogenic agents for the therapy of local and metastatic human tumors. Cancer Gene Therapy ( 2001 ) 8, 45 ± 54
Gene therapy prospects--intranasal delivery of therapeutic genes
Advances in clinical and experimental medicine : official organ Wroclaw Medical University
Gene therapy is recognized to be a novel method for the treatment of various disorders. Gene therapy strategies involve gene manipulation on broad biological processes responsible for the spreading of diseases. Cancer, monogenic diseases, vascular and infectious diseases are the main targets of gene therapy. In order to obtain valuable experimental and clinical results, sufficient gene transfer methods are required. Therapeutic genes can be administered into target tissues via gene carriers commonly defined as vectors. The retroviral, adenoviral and adeno-associated virus based vectors are most frequently used in the clinic. So far, gene preparations may be administered directly into target organs or by intravenous, intramuscular, intratumor or intranasal injections. It is common knowledge that the number of gene therapy clinical trials has rapidly increased. However, some limitations such as transfection efficiency and stable and long-term gene expression are still not resolved. Co...
Surgery, 2001
Background. Many gene therapy strategies would benefit from efficient, regional organ delivery of therapeutic genes. Methods. Regional perfusions of lung, liver, or bladder were performed to determine if rapid and efficient gene transfer can be accomplished in vivo, and to determine if in vivo gene transfer can be limited to the organ of interest. In addition, herpes simplex virus tumor necrosis factor (HSVtnf), carrying the human tumor necrosis factor α gene was used as a treatment for methylcholanthrene sarcoma in a syngeneic lung metastases model in Fisher rats.