Jeschke M G, Klein D
Plastic and Hand Surgery, University of Erlangen, University Hospital, Krankenhausstrasse 12, 91054 Erlangen, Germany.
Gene Ther. 2004 May;11(10):847-55. doi: 10.1038/sj.gt.3302229.
Liposomal gene transfer is an effective therapeutic approach for the treatment of several pathophysiologic states. The purpose of the present study was to define whether gene transfer of multiple genes is a feasible approach and whether this approach would be more effective than the single transfer of cDNA. Rats were inflicted an acute wound and divided into four groups to receive weekly subcutaneous injections of liposomes plus the Lac-Z gene (0.22 microg, vehicle), or liposomes plus the insulin like-growth factor-I (IGF-I)cDNA (2.2 microg) and Lac Z gene (0.22 microg), or liposomes plus the keratinocyte growth factor (KGF) cDNA (2.2 microg) and Lac Z gene (0.22 microg), or liposomes plus the IGF-I/KGF cDNA (2.2 microg) and Lac Z gene (0.22 microg). Planimetry, immunological assays, histological and immunohistochemical techniques were used to determine molecular mechanisms after gene transfer, protein expression, dermal and epidermal regeneration. IGF-I/KGF cDNA transfer increased IGF-I and KGF protein concentration and caused concomitant cellular responses, for example,by increasing IGFBP-3, P<0.05. IGF-I/KGF cDNA gene transfer improved epidermal regeneration by exhibiting the most rapid area and linear wound re-epithelization by almost 250% compared to control and each growth factor given individually, P<0.001, which was probably due to promitogenic and antiapoptotic effects on basal keratinocytes when compared to controls, P<0.001. Dermal regeneration was improved in IGF-I/KGF cDNA-treated animals by an increased collagen deposition and morphology when compared with vehicle, IGF-I and KGF, P<0.001. IGF-I/KGF cDNA increased VEGF concentrations and thus neovascularization when compared with vehicle, IGF-I and KGF, P<0.001. In the present study, we showed that exogenous gene transfer of multiple cDNA sequences have an additive effect on intracellular and biological responses when compared to the same gene administered as a single cDNA sequence. Our findings demonstrate that gene therapy with multiple genes is feasible, and that the gene transfer of multiple genes can enhance and accelerate physiologic and biological effects.
脂质体基因转移是治疗多种病理生理状态的一种有效治疗方法。本研究的目的是确定多基因转移是否是一种可行的方法，以及这种方法是否比单独转移互补DNA（cDNA）更有效。给大鼠造成急性伤口，并将其分为四组，每周皮下注射脂质体加Lac-Z基因（0.22微克，赋形剂），或脂质体加胰岛素样生长因子-I（IGF-I）cDNA（2.2微克）和Lac Z基因（0.22微克），或脂质体加角质形成细胞生长因子（KGF）cDNA（2.2微克）和Lac Z基因（0.22微克），或脂质体加IGF-I/KGF cDNA（2.2微克）和Lac Z基因（0.22微克）。采用平面测量、免疫测定、组织学和免疫组织化学技术来确定基因转移后的分子机制、蛋白质表达、真皮和表皮再生情况。IGF-I/KGF cDNA转移增加了IGF-I和KGF蛋白浓度，并引发了相应的细胞反应，例如，通过增加IGFBP-3，P<0.05。与对照组以及单独给予的每种生长因子相比，IGF-I/KGF cDNA基因转移通过展现出最快的面积和线性伤口再上皮化，使表皮再生提高了近250%，P<0.001，这可能是由于与对照组相比，其对基底角质形成细胞具有促有丝分裂和抗凋亡作用，P<0.001。与赋形剂、IGF-I和KGF相比，IGF-I/KGF cDNA处理的动物真皮再生通过增加胶原蛋白沉积和改善形态而得到改善，P<0.001。与赋形剂、IGF-I和KGF相比，IGF-I/KGF cDNA增加了血管内皮生长因子（VEGF）浓度，从而促进了新血管形成，P<0.001。在本研究中，我们表明，与作为单一cDNA序列给予的相同基因相比，多个cDNA序列的外源基因转移对细胞内和生物学反应具有累加效应。我们的研究结果表明，多基因治疗是可行的，并且多基因转移可以增强和加速生理和生物学效应。
