Targeted Delivery of siRNA to Transferrin Receptor Overexpressing Tumor Cells via Peptide Modified Polyethylenimine

Abstract

Abstract:The use of small interference RNA (siRNA) to target oncogenes is a promising treatment approach for cancer. However, siRNA cancer therapies are hindered by poor delivery of siRNA to cancer cells. Transferrin receptor (TfR) is overexpressed in many types of tumor cells and therefore is a potential target for the selective delivery of siRNA to cancer cells. Here, we used the TfR binding peptide HAIYPRH (HAI peptide) conjugated to cationic polymer branched polyethylenimine (bPEI), optimized the coupling strategy, and the TfR selective delivery of siRNA was evaluated in cells with high (H1299) and low TfR expression (A549 and H460). The HAI-bPEI conjugate exhibited chemico-physical properties in terms of size, zeta-potential, and siRNA condensation efficiency similar to unmodified bPEI. Confocal microscopy and flow cytometry results revealed that HAI-bPEI selectively delivered siRNA to H1299 cells compared with A549 or H460 cells. Moreover, HAI-bPEI achieved more efficient glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene knockdown in H1299 cells compared with bPEI alone. However, despite optimization of the targeting peptide and coupling strategy, HAI-bPEI can only silence reporter gene enhanced green fluorescent protein (eGFP) at the protein level when chloroquine is present, indicating that further optimization of the conjugate is required. In conclusion, the HAI peptide may be useful to target TfR overexpressing tumors in targeted gene and siRNA delivery approaches.

Construction and Biological Evaluation of a Novel Integrin ανβ3-Specific Carrier for Targeted siRNA Delivery In Vitro

“…The peptides of c(RGD) 2 -9R recognize and interact with integrin by RGD sequence, and provide the ability of targeting to a variety of cancer cells where α v β 3 integrin were overexpressed. Conjugating the carrier with a targeted or functional moiety is a useful and popular strategy for siRNA delivery, and has been validated in many preclinical studies [ 22 , 23 , 24 ].…”

Blocking transferrin receptor inhibits the growth of lung adenocarcinoma cells in vitro

“…Our investigation of the functions and properties of TfR in iron transport indicate that TfR plays a greater biological role than previously thought . Xie et al reported that H1299 cells demonstrate significantly higher TfR expression compared to A549 and H460 cells . Therefore, we chose to use A549 and H1299 cell lines to locate a TfR overexpressing cell.…”

“…5,20,21 Xie et al reported that H1299 cells demonstrate significantly higher TfR expression compared to A549 and H460 cells. 22 Therefore, we chose to use A549 and H1299 cell lines to locate a TfR overexpressing cell. In this study, we observed upregulated protein levels of cellular TfR in lung adenocarcinoma H1299 cells (Fig 1a,b).…”

Noninvasive imaging of c(RGD)2‐9R as a potential delivery carrier for transfection of siRNA in malignant tumors

“…The peptide of cyclic (RGD) 2 targeting integrin α ν β 3 was conjugated with 9R, designated as c(RGD) 2 ‐9R. The control peptide of cyclic (GGG) 2 , which did not target any known receptors of tumors, was also conjugated with 9R, designated as c(GGG) 2 ‐9R. All the peptides were synthesized by ChinaPeptides Co, Ltd, China.…”