Abstract
Cationic liposome-DNA complex (CLDC)-based intravenous gene delivery targets gene expression to vascular endothelial cells, macrophages and tumor cells. We used systemic gene delivery to identify anti-angiogenic gene products effective against metastatic spread in tumor-bearing mice. Specifically, CLDC-based intravenous delivery of the p53 and GM-CSF genes were each as effective as the potent antiangiogenic gene, angiostatin, in reducing both tumor metastasis and tumor angiogenesis. Combined delivery of these genes did not increase anti-tumor activity, further suggesting that each gene appeared to produce its antimetastatic activity through a common antiangiogenic pathway. CLDC-based intravenous delivery of the human wild type p53 gene transfected up to 80% of tumor cells metastatic to lung. Furthermore, it specifically induced the expression of the potent antiangiogenic gene, thrombospondin-1, indicating that p53 gene delivery in vivo may inhibit angiogenesis by inducing endogenous thrombospondin-1 expression. CLDC-based delivery also identified a novel anti-tumor activity for the metastasis suppressor gene CC3. Thus, CLDC-based intravenous gene delivery can produce systemic antiangiogenic gene therapy using a variety of different genes and may be used to assess potential synergy of combined anti-tumor gene delivery and to identify novel activities for existing anti-tumor genes.
Highlights
Cationic liposome-DNA complex (CLDC)-based intravenous gene delivery targets gene expression to vascular endothelial cells, macrophages and tumor cells
We show that these redesigned CLDC are able to transfect large numbers of metastatic tumor cells with the wild type human p53 gene following intravenous injection into C57Bl6 mice bearing metastatic B16-F10 melanoma tumors
Individual mice in groups of eight received 400 nmol of DOTIM: cholesterol multilamellar vesicles (MLV) complexed to 25 g of an HCMV-driven expression plasmid containing the murine angiostatin gene, the murine GM-CSF gene, the human wild type p53 gene, or the CAT reporter gene on day 3 and again on day 10 following tumor inoculation
Summary
Plasmids p4199 —CMV-p53 was constructed by isolating a 1.4-kilobase pair HindIII–SmaI fragment containing the wild type human p53 cDNA Vogelstein) and ligating it into the HindIII–SmaI site of p4136 [11]. P4305—CMV-angiostatin was constructed by isolating a 1.4-kilobase pair HindIII–XbaI fragment containing the murine angiostatin cDNA P4442—CMV-BCL-2 was constructed by ligating the human BCL-2 cDNA Korsmeyer) into the EcoRV site of pVR1223 [12]. P4447—CMV-CC3 was obtained by isolating a 0.8-kilobase pair fragment containing the human CC3 and ligating it into the EcoRV site of pVR1223. Transferase (CAT) [11], and p4241, CMV-luciferase [8], have been described. Plasmids were purified as described previously [11]
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