Tumor Imaging and Interferon-γ–Inducible Protein-10 Gene Transfer Using a Highly Efficient Transferrin-Conjugated Liposome System in Mice

Abstract

Purpose: We have developed a PEGylated transferrin-conjugated liposomes (PTf-Ls) system for the combined tumor imaging and targeted delivery of the IFN-g-inducible protein-10 (IP-10) gene in a single macromolecular construct. Here, we characterize and analyze the use of this system in a mouse model of breast cancer.Experimental Design: The biophysical and cell transfection properties of PTf-Ls were determined through a series of in vitro experiments. A nude mouse/breast cancer cell line xenograft model (mouse xenograft model) was used to image the tumor internalization of fluorescently labeled PTf-Ls. The clinical use of the system was tested by treating tumor-bearing mice with PTf-Ls loaded with IP-10 plasmid DNA or fluorescent lipoplexes.Results: The resulting 165-nm liposomes (zeta potential ¼ À10.6 mV) displayed serum resistance, low cytotoxicity (<5%), and high transfection efficiency ( 82.8%) in cultured cells. Systemic intravenous administration of fluorescent PTf-Ls in the mouse xenograft model resulted in nanoparticle circulation for 72 hours, as well as selective and efficient internalization in tumor cells, according to in vivo fluorescence and bioluminescence analyses. Tumor fluorescence increased gradually up to 26 hours, whereas background fluorescence decreased to near-baseline levels. Treatment of mice with PTf-Ls entrapped pcDNA3.1-IP-10 suppressed tumor growth in mice by 79% on day 50 and increased the mean survival time of mice. Fluorescent pcDNA-IP-10-entrapped PTf-Ls showed good properties for simultaneous tumor-targeted imaging and gene-specific delivery in an animal tumor model.

Reduced temporal sampling effect on accuracy of time-domain fluorescence lifetime Förster resonance energy transfer

“…More precisely, we employed a NIR FRET transferrin (Tfn) assay in 96-well plate settings. Tfn has been used as a carrier for anticancer drugs or other therapeutic agents [44][45][46] to enhance the internalization specificity into neoplastic tissues. The Tfn receptor (TfnR) is homodimeric, i.e., two molecules of Tfn bind within 2 to 10 nm, which allows the use of FRET-based imaging techniques.…”

Efficient targeted tumor imaging and secreted endostatin gene delivery by anti-CD105 immunoliposomes

“…Liposome (Lp) is versatile carrier systems for the delivery of cytotoxic drugs or genes, and provides a platform for enhancing the efficiency of delivery and targeting of therapies to tumor tissues [ 11 – 16 ] PEGylated Lp, in which polyethylene glycol lipids are incorporated into the lipid bilayer, have the advantage of preventing loaded gene degradation and clearance [ 12 ]. Furthermore, when combined with tumor-specific markers, the efficiency of targeted delivery can be significantly enhanced [ 11 , 13 ]. Nanographenes synthesized to target CD105 with anti-CD105 mAb or microbubbles conjugated with anti-vascular endothelial growth factor (VEGF) mAb have been reported to be specifically directed to tumor microvasculature in vivo [ 15 , 16 ].…”

“…Nanographenes synthesized to target CD105 with anti-CD105 mAb or microbubbles conjugated with anti-vascular endothelial growth factor (VEGF) mAb have been reported to be specifically directed to tumor microvasculature in vivo [ 15 , 16 ]. The significantly improved tumor targeting efficiency of a transferrin-conjugated Lp system described in our previous work [ 13 ] may be utilized for tumor neovasculature-targeted imaging and/or therapeutic gene delivery in vivo. To the best of our knowledge, little has been reported to date about targeted CD105 Lp delivery systems incorporating both anticancer genes and fluorescent dyes in different compartments of a single vesicle.…”

Folate-modified Chitosan Nanoparticles Containing the IP-10 Gene Enhance Melanoma-specific Cytotoxic CD8+CD28+ T Lymphocyte Responses

“…IP-10 is thought to negatively regulate angiogenesis through directly inhibiting the expression of angiogenic factors as well as activating of monocytes, fibrocytes, epithelial cells, and keratinocytes 31 , 32 . Moreover, IP-10 plays a role in cell proliferation and apoptosis 33 , 34 .…”