Enhancing Anticancer Effect of Gefitinib across the Blood–Brain Barrier Model Using Liposomes Modified with One α-Helical Cell-Penetrating Peptide or Glutathione and Tween 80

Abstract

Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKI), such as gefitinib, have been demonstrated to effectively treat the patients of extracranial non-small cell lung cancer (NSCLC). However, these patients often develop brain metastasis (BM) during their disease course. The major obstacle to treat BM is the limited penetration of anticancer drugs across the blood–brain barrier (BBB). In the present study, we utilized gefitinib-loaded liposomes with different modifications to improve gefitinib delivery across the in vitro BBB model of bEnd.3 cells. Gefitinib was encapsulated in small unilamellar liposomes modified with glutathione (GSH) and Tween 80 (SUV-G+T; one ligand plus one surfactant) or RF (SUV-RF; one α-helical cell-penetrating peptide). GSH, Tween 80, and RF were tested by the sulforhodamine B (SRB) assay to find their non-cytotoxic concentrations on bEnd.3 cells. The enhancement on gefitinib across the BBB was evaluated by cytotoxicity assay on human lung adenocarcinoma PC9 cells under the bEnd.3 cells grown on the transwell inserts. Our findings showed that gefitinib incorporated in SUV-G+T or SUV-RF across the bEnd.3 cells significantly reduced the viability of PC9 cells more than that of free gefitinib. Furthermore, SUV-RF showed no cytotoxicity on bEnd.3 cells and did not affect the transendothelial electrical resistance (TEER) and transendothelial permeability of sodium fluorescein across the BBB model. Moreover, flow cytometry and confocal laser scanning microscopy were employed to evaluate the endocytosis pathways of SUV-RF. The results indicated that the uptake into bEnd.3 cells was mainly through adsorptive-mediated mechanism via electrostatic interaction and partially through clathrin-mediated endocytosis. In conclusion, cell penetrating peptide-conjugated SUV-RF shed light on improving drug transport across the BBB via modulating the transcytosis pathway(s).

Stealth and Cationic Nanoliposomes as Drug Delivery Systems to Increase Andrographolide BBB Permeability

“…Liposomes could be internalized into cells by different mechanisms, according to the type of cell, composition, surface charge, and size of the liposome [ 45 , 46 , 47 ]. In our study, the cellular association of the liposomes was significantly influenced by indomethacin, with a reduction in cellular association of about 44% and 63% for LPs-6C and CLPs-6C, respectively ( Figure 7 ).…”

Brain Metastasis Treatment: The Place of Tyrosine Kinase Inhibitors and How to Facilitate Their Diffusion across the Blood–Brain Barrier

“…One formulation combined glutathione and polysorbate 80, known to be two ligands of physiological receptors in the blood–brain barrier, and another formulation used liposomes functionalized with RF, a cell-penetrating peptide. Both formulations showed an enhanced uptake across bEnd.3 cells, partially through clathrin-mediated endocytosis for RF-liposomes [ 115 ]. None of these molecules has been approved by the FDA to date.…”

A journey from the endothelium to the tumor tissue: distinct behavior between PEO-PCL micelles and polymersomes nanocarriers

“…( 2008 ) observed translocation of solid lipid nanoparticles through endothelial cells from bovine aortas. Zhao and Lin worked on endothelial layers from blood-brain barrier, for Zhao, there was a possible translocation of the nanoparticles, or for Lin, at least of the nanocarriers charge (Lin et al., 2016 ; Zhao et al., 2016 ). The increase in DiO’s passage induced by a cytokine or cancer cells was more expected.…”

“…For the model of translocation across endothelium, an adaptation of transwell assays (on Boyden chambers) was used. Well-documented for chemotaxis, transwell assays only start to be used to study the passage of nanomaterials across biological barriers such as mucus, aortic, brain, and intestinal barriers (Broughton-Head et al., 2007 ; Jayagopal et al., 2008 ; Cohen et al., 2014 ; Shah et al., 2015 ; Kasper et al., 2016 ; Lin et al., 2016 ; Peuschel et al., 2016 ; Zhao et al., 2016 ; Boya et al., 2017 ; Hsiao et al., 2017 ). So far, this promising and flexible technique had not been used with polymeric nanocarriers or to mimic the endothelium.…”