Lipoplexes with alkylphospholipid as new helper lipid for efficient in vitro and in vivo gene transfer in tumor therapy

Abstract

To improve liposomal gene transfer we investigated the influence of membrane-interacting alkylphospholipids (APLs) on gene transfer efficiency in vitro and in vivo using the LacZ reporter gene and the cytosine deaminase (CD) suicide gene. Liposomes were first optimized concerning the kind and amount of APL and the additional liposome components. Thus, an up to 270% increase in the transfer efficiency of the LacZ gene into HCT15 and HCT116 human colon carcinoma cells could be obtained in vitro compared to lipofectin-mediated transfection by using a lipoplex consisting of tetradecylphosphocholine/dimethyldioctadecylamine/ cholesterol/dioleylphosphoethanolamine-liposomes and the pSV40-bGal-plasmid. The in vivo experiments revealed that alkylphospholipid-lipoplexes (APL-LPs) were similarly effective in the transfer of the LacZ gene into colon carcinoma as formulations consisting of lipofectin. Using the CD-gene in combination with APL-LPs resulted in a significantly stronger inhibition of C26 colon carcinoma growth compared to lipofectin-mediated gene transfer following treatment of mice with the prodrug 5-fluorocytosine. The results of this study demonstrate for the first time that the utilization of membrane-active APLs as component of the liposomal part of lipoplexes enhances the efficacy of gene therapy in vitro and in vivo.

Comparison of Transfection Efficiency of Nonviral Gene Transfer Reagents

“…Other tumor cell lines, HeLa cervical cancer, Hep G2 hepatocellular carcinoma, and HCT116 colorectal carcinoma, have been used for transfection studies with PEI [38,39], cationic lipids [40], and cationic liposome [41,42]. In the present study, the highest transfection efficiency in HeLa cells using both luciferase and b-galactosidase genes in the presence of serum was seen with FuGENE HD and in the absence of serum was seen with Arrest-In.…”

Erufosine (ErPC3) Cationic Prodrugs as Dual Gene Delivery Reagents for Combined Antitumor Therapy

“…In 2003, Zeisig et al. described gene transfer with liposomal formulations incorporating APLs as helper lipids, assuming that APLs may promote the transmembrane transport of the plasmid‐lipid complex (lipoplex) into the target cell due to their “detergent‐like properties”. Introduction of 2 % tetradecylphosphocholine (TPC), a lower carbon homologue of miltefosine, in the lipoplexes improved transfer efficiency of the LacZ gene into HCT15 and HCT116 human colon carcinoma cells up to 43 %.…”

“…[12] Furthermore, some of the pathways by which APLs lead to cytotoxicity have been shown to influence radiosensitivity or radioresistance of tumor cells, which resulted in successful clinicalo utcomesf or the combination of APLs with radiotherapy. [13] On the other hand, there are only very few reports on the combinationo fA PLs with cancerg ene therapy.I n2 003, Zeisig et al described gene transfer with liposomal formulations incorporating APLs as helper lipids, [14] assumingt hat APLs may promote the transmembrane transport of the plasmid-lipid complex (lipoplex) into the target cell due to their "detergent-like properties". Introduction of 2% tetradecylphosphocholine (TPC), al ower carbon homologue of miltefosine, in the lipoplexesi mproved transfer efficiency of the LacZ gene into HCT15 and HCT116 human colon carcinoma cells up to 43 %.…”

A glycobiology review: Carbohydrates, lectins and implications in cancer therapeutics

“…Therapeutic agents are typically linked to the DDS via a biodegradable spacer such as the tetrapeptide Gly-Phe-Leu-Gly which is digested by the enzymatic activity of cathepsin B, thus liberating the therapeutic agent (Kopecek et al, 2000; Kopecek et al, 2001; Lu et al, 2002). Alternatively, the entire DDS may be biodegradable within the target cell (Zeisig et al, 2003). An example of this system would be the combination of horseradish peroxidase with indole-3-acetic acid.…”