Sterically stabilized BGTC-based lipoplexes: structural features and gene transfection into the mouse airwaysin vivo

Pitard, Bruno; Oudrhiri, Noufϊssa; Lambert, Olivier; Vivien, Eric; Masson, Christophe; Wetzer, Barbara; Hauchecorne, M.; Scherman, Daniel; Rigaud, Jean‐Louis; Vigneron, Jean‐Pierre; Lehn, Jean‐Maríe; Lehn, Pierre

doi:10.1002/jgm.211

Cited by 61 publications

(79 citation statements)

References 32 publications

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“…24 In addition, these vectors conjuguated to poly(ethyleneglycol) could be sterically stabilized for in vivo use. 30 Further studies are needed to explore these alternative gene therapy strategies for HCC.…”

Section: Discussionmentioning

confidence: 99%

“…This procedure should be better sterically stabilized by the addition of modified poly(ethyleneglycol) (Chol-PEG), as soon as we can have access to this patented compound. 30 Macroscopic analyses of tumors treated with either TIMP-2 or TIMP-3 showed a disintegrated white appearance of the tumors (not shown). However, a hemorrhaged and hypervascularized aspect was noted in all tumors treated with pSK.CMVnlslacZ (not shown).…”

Section: Treatment Of Established Huh7 Tumors By Repeated Injections mentioning

confidence: 98%

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Gene therapy for hepatocellular carcinoma using non-viral vectors composed of bis guanidinium-tren-cholesterol and plasmids encoding the tissue inhibitors of metalloproteinases TIMP-2 and TIMP-3

et al. 2003

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Metalloproteinases (MMPs) and their natural inhibitors (TIMPs) contribute to the regulation of tumor microenvironment. Their expressions are deregulated in almost all human cancers. We report a novel approach to gene therapy of hepatocellular carcinoma (HCC), using repeated injections of DNA plasmids encoding the tissue inhibitors of metalloproteinases (TIMPs) TIMP-2 or TIMP-3, and a novel competent formulation of gene transfer based on nontoxic cationic cholesterol derivatives. The new gene delivery system was efficient in demonstrating the antitumor efficiency of TIMP-2 or TIMP-3 in inhibiting tumor growth of human HuH7 HCC cells xenografted into nude mice. We show, for the first time, an in vivo effect of TIMP-3 in delaying HCC tumor growth. No treatment-related toxicity was noted. An inhibition of angiogenesis and tumor necrosis accompanied the inhibitory effects of TIMP-2 or TIMP-3 on tumor expansion and invasion. We also report a bystander effect produced by transfected HuH7 tumor cells mixed with untransfected cells in 1:1 ratio in culture that resulted in killing 98% of cells within 96 h. In addition, the soluble forms of TIMP-2 and TIMP-3 expressed by transfected cells exerted a cytotoxic effect on untransfected HuH7 cell cultures. Taken together, these results demonstrate the potential efficacy of repeated treatment of secreted TIMP-2 and TIMP-3 for the design of nonviral gene therapy for hepatocarcinoma.

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Section: Discussionmentioning

confidence: 99%

Section: Treatment Of Established Huh7 Tumors By Repeated Injections mentioning

confidence: 98%

Gene therapy for hepatocellular carcinoma using non-viral vectors composed of bis guanidinium-tren-cholesterol and plasmids encoding the tissue inhibitors of metalloproteinases TIMP-2 and TIMP-3

et al. 2003

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“…Ever since Felgner et al 3 reported their pioneering development of cationic lipid-mediated gene delivery protocol in 1987, many reports aimed at developing more efficient cationic transfection lipids have appeared in the literature. [4][5][6][7][8][9][10] Recently, we have also developed efficient nonglycerol-based cationic transfection lipids. [11][12][13][14][15] In sharp contrast, details of cationic lipid-mediated transfection pathways still remain elusive.…”

Section: Introductionmentioning

confidence: 99%

Single histidine residue in head-group region is sufficient to impart remarkable gene transfection properties to cationic lipids: evidence for histidine-mediated membrane fusion at acidic pH

et al. 2003

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Presence of endosome-disrupting multiple histidine functionalities in the molecular architecture of cationic polymers, such as polylysine, has previously been demonstrated to significantly enhance their in vitro gene delivery efficiencies. Towards harnessing improved transfection property through covalent grafting of endosome-disrupting single histidine functionality in the molecular structure of cationic lipids, herein, we report on the design, the synthesis and the transfection efficiency of two novel nonglycerol-based histidylated cationic amphiphiles. We found that L-histidine-(N,N-di-n-hexadecylamine)ethylamide (lipid 1) and L-histidine-(N,N-di-n-hexadecylamine,-N-methyl)ethylamide (lipid 2) in combination with cholesterol gave efficient transfections into various cell lines. The transfection efficiency of Chol/lipid 1 lipoplexes into HepG2 cells was two order of magnitude higher than that of FuGENE TM 6 and DC-Chol lipoplexes, whereas it was similar into A549, 293T7 and HeLa cells. A better efficiency was obtained with Chol/lipid 2 lipoplexes when using the cytosolic luciferase expression vector (pT7Luc) under the control of the bacterial T7 promoter. Membrane fusion activity measurements using fluorescence resonance energy transfer (FRET) technique showed that the histidine head-groups of Chol/lipid 1 liposomes mediated membrane fusion in the pH range 5-7. In addition, the transgene expression results using the T7Luc expression vector convincingly support the endosome-disrupting role of the presently described mono-histidylated cationic transfection lipids and the release of DNA into the cytosol. We conclude that covalent grafting of a single histidine amino acid residue to suitable twin-chain hydrophobic compounds is able to impart remarkable transfection properties on the resulting mono-histidylated cationic amphiphile, presumably via the endosome-disrupting characteristics of the histidine functionalities.

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“…It should be stressed here that we have already reported that incorporation of appropriate lipophilic PEG derivatives allowed to prepare highly concentrated solutions of colloidally stable BGTC/DOPE lipoplexes that permitted enhanced transfection of the mouse airways in vivo via intranasal instillation. 34 In addition, we have recently observed that incorporation of such lipophilic PEG derivatives allows efficient gene transfection into the mouse respiratory epithelium when using aminoglycosidederived cationic lipids. 39 Our results also indicate that the various transgenes (HSV-TK, EGFP, LacZ) were preferentially expressed in the peritoneal tumor tissues.…”

Section: Discussionmentioning

confidence: 99%

“…30 We also found that BGTC/DOPE liposomes mediate efficient gene transfection into the mouse respiratory epithelium in vivo, the lipoplexes being instilled either directly into the trachea or into the nasal cavity. 33,34 The aim of the present work was thus to assess the therapeutic potential of a novel nonviral gene therapy approach consisting of BGTC-mediated in vivo lipofection of a combined suicide gene system for the treatment of pancreatic peritoneal carcinomatosis. I.p.…”

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confidence: 99%

Combined suicide gene therapy for pancreatic peritoneal carcinomatosis using BGTC liposomes

et al. 2003

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Peritoneal dissemination is a common end-stage complication of pancreatic cancer for which novel therapeutic modalities are actively investigated, as there is no current effective therapy. Thus, we evaluated, in a mouse model of pancreatic peritoneal carcinomatosis, the therapeutic potential of a novel nonviral gene therapy approach consisting of bis-guanidinium-tren-cholesterol (BGTC)-mediated lipofection of a combined suicide gene system. Human BxPC-3 pancreatic cells secreting the carcinoembryonic antigen (CEA) tumor marker were injected into the peritoneal cavity of nude mice. After 8 days, intraperitoneal (i.p.) lipofection was performed using BGTC/DOPE cationic liposomes complexed with plasmids encoding the two prodrug-activating enzymes Herpes Simplex Virus thymidine kinase and Escherichia coli cytosine deaminase, the latter being expressed from a bicistronic cassette also encoding E. coli uracil phosphoribosyltransferase. Administration of the lipoplexes was followed by treatment with the corresponding prodrugs ganciclovir and 5-fluorocytosine. The results presented herein demonstrate that BGTC/DOPE liposomes can efficiently mediate gene transfection into peritoneal tumor nodules. Indeed, HSV-TK mRNA was detected in tumor nodule tissues by semiquantitative reverse transcription-polymerase chain reaction analysis. In addition, green fluorescent protein (GFP) fluorescence and X-gal staining were observed in the peritoneal tumor foci following lipofection of the corresponding EGFP and LacZ reporter genes. These expression analyses also showed that transgene expression lasted for about 2 weeks and was preferential for the tumor nodules, this tumor preference being in good agreement with the absence of obvious treatment-related toxicity. Most importantly, mice receiving the full treatment scheme (BGTC liposomes, suicide genes and prodrugs) had significantly lower serum CEA levels than those of the various control groups, a finding indicating that peritoneal carcinomatosis progression was strongly reduced in these mice. In conclusion, our results demonstrate the therapeutic efficiency of BGTC-mediated i.p. lipofection of a combined suicide gene system in a mouse peritoneal carcinomatosis model and suggest that BGTC-based prodrug-activating gene therapy approaches may constitute a potential treatment modality for patients with peritoneal carcinomatosis and minimal residual disease.

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Sterically stabilized BGTC-based lipoplexes: structural features and gene transfection into the mouse airwaysin vivo

Abstract: Our results represent an important step towards the design of multimodular BGTC-based systems for improved in vivo gene transfection.

Cited by 61 publications

References 32 publications

Gene therapy for hepatocellular carcinoma using non-viral vectors composed of bis guanidinium-tren-cholesterol and plasmids encoding the tissue inhibitors of metalloproteinases TIMP-2 and TIMP-3

Gene therapy for hepatocellular carcinoma using non-viral vectors composed of bis guanidinium-tren-cholesterol and plasmids encoding the tissue inhibitors of metalloproteinases TIMP-2 and TIMP-3

Single histidine residue in head-group region is sufficient to impart remarkable gene transfection properties to cationic lipids: evidence for histidine-mediated membrane fusion at acidic pH

Combined suicide gene therapy for pancreatic peritoneal carcinomatosis using BGTC liposomes

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