de Wilt J H, Bout A, Eggermont A M, van Tiel S T, de Vries M W, ten Hagen T L, de Roos W K, Valerio D, van der Kaaden M E
Department of Surgical Oncology, University Hospital Rotterdam Dijkzigt/Daniel de Hoed Cancer Center, Rotterdam, The Netherlands.
Hum Gene Ther. 2001 Mar 20;12(5):489-502. doi: 10.1089/104303401300042384.
Cytokine gene transfer using (multiple) intratumoral injections can induce tumor regression in several animal models, but this administration technique limits the use for human gene therapy. In the present studies we describe tumor growth inhibition of established limb sarcomas after a single isolated limb perfusion (ILP) with recombinant adenoviral vectors harboring the rat IL-3 beta gene (IG.Ad.CMV.rIL-3 beta). In contrast, a single intratumoral injection or intravenous administration did not affect tumor growth. Dose-finding studies demonstrated a dose-dependent response with a loss of antitumor effect below 1 x 10(9) IU of IG.Ad.CMV.rIL-3 beta. Perfusions with adenoviral vectors bearing a weaker promoter (MLP promoter) driving the rIL-3 beta gene did not result in antitumor responses, suggesting that the rIL-3 beta-mediated antitumor effect depends on the amount of rIL-3 beta protein expressed by the infected cells. Furthermore, it was shown by direct comparison that ILP with IG.Ad.CMV.rIL-3 beta in the ROS-1 osteosarcoma model is at least as efficient as the established therapy with the combination of TNF-alpha and melphalan. Treatment with IG.Ad.CMV.rIL-3 beta induced a transient dose-dependent leukocytosis accompanied by an increase in peripheral blood levels of histamine. Leukocyte infiltrations were also histopathologically demonstrated in tumors after perfusion. These results demonstrate that ILP with recombinant adenoviral vectors carrying the IL-3 beta transgene inhibits tumor growth in rats and suggest that cytokine gene therapy using this administration technique might be beneficial for clinical cancer treatment.
在多种动物模型中，通过（多次）瘤内注射进行细胞因子基因转移可诱导肿瘤消退，但这种给药技术限制了其在人类基因治疗中的应用。在本研究中，我们描述了用携带大鼠IL-3β基因（IG.Ad.CMV.rIL-3β）的重组腺病毒载体进行单次孤立肢体灌注（ILP）后，已建立的肢体肉瘤的肿瘤生长受到抑制。相比之下，单次瘤内注射或静脉内给药对肿瘤生长没有影响。剂量探索研究表明存在剂量依赖性反应，当IG.Ad.CMV.rIL-3β剂量低于1×10⁹ IU时，抗肿瘤作用丧失。用驱动rIL-3β基因的较弱启动子（MLP启动子）的腺病毒载体进行灌注未产生抗肿瘤反应，这表明rIL-3β介导的抗肿瘤作用取决于感染细胞表达的rIL-3β蛋白量。此外，通过直接比较表明，在ROS-1骨肉瘤模型中用IG.Ad.CMV.rIL-3β进行ILP至少与已确立的TNF-α和美法仑联合治疗一样有效。用IG.Ad.CMV.rIL-3β治疗诱导了短暂的剂量依赖性白细胞增多，并伴有外周血组胺水平升高。灌注后肿瘤组织病理学检查也证实有白细胞浸润。这些结果表明，用携带IL-3β转基因的重组腺病毒载体进行ILP可抑制大鼠肿瘤生长，并表明使用这种给药技术的细胞因子基因治疗可能对临床癌症治疗有益。
