pubmed.ncbi.nlm.nih.gov

Nanostructured lipid carriers as multifunctional nanomedicine platform for pulmonary co-delivery of anticancer drugs and siRNA - PubMed

  • ️Tue Jan 01 2013

Nanostructured lipid carriers as multifunctional nanomedicine platform for pulmonary co-delivery of anticancer drugs and siRNA

Oleh Taratula et al. J Control Release. 2013.

Abstract

We developed, synthesized, and tested a multifunctional nanostructured lipid nanocarrier-based system (NLCS) for efficient delivery of an anticancer drug and siRNA directly into the lungs by inhalation. The system contains: (1) nanostructured lipid carriers (NLC); (2) anticancer drug (doxorubicin or paclitaxel); (3) siRNA targeted to MRP1 mRNA as a suppressor of pump drug resistance; (4) siRNA targeted to BCL2 mRNA as a suppressor of nonpump cellular resistance and (5) a modified synthetic analog of luteinizing hormone-releasing hormone (LHRH) as a targeting moiety specific to the receptors that are overexpressed in the plasma membrane of lung cancer cells. The NLCS was tested in vitro using human lung cancer cells and in vivo utilizing mouse orthotopic model of human lung cancer. After inhalation, the proposed NLCS effectively delivered its payload into lung cancer cells leaving healthy lung tissues intact and also significantly decreasing the exposure of healthy organs when compared with intravenous injection. The NLCS showed enhanced antitumor activity when compared with intravenous treatment. The data obtained demonstrated high efficiency of proposed NLCS for tumor-targeted local delivery by inhalation of anticancer drugs and mixture of siRNAs specifically to lung cancer cells and, as a result, efficient suppression of tumor growth and prevention of adverse side effects on healthy organs.

Keywords: Drug resistance; Imaging; Inhalation; Luteinizing hormone-releasing hormone (LHRH); Nanostructured lipid carrier; Orthotopic lung cancer model.

Copyright © 2013 Elsevier B.V. All rights reserved.

PubMed Disclaimer

Figures

Figure 1
Figure 1

A schematic representation of Nanostructured Lipid Carrier (NLC)-based drug delivery system for pulmonary co-delivery of an anticancer drug, siRNA and targeting peptide.

Figure 2
Figure 2

Characterization of LHRH-targeted paclitaxel (TAX)-loaded Nanostructured Lipid Carriers (LHRH-NLC-TAX-siRNA). (A) Size distribution of LHRH-NLC-TAX-siRNA measured by dynamic light scattering before and after nebulization. (B) Atomic Force Microscope image of LHRH-NLC-TAX-siRNA.

Figure 3
Figure 3

Complexation of siRNA with NLC. (A) A representative agarose gel electrophoresis image illustrating siRNA complexation efficiency by LHRH-NLC-TAX at the following w/w ratio (weight NLC/weight siRNA): (1) 0:1 (naked siRNA, control); (2) 10:1; (3) 50:1; (4) 100:1; and (5) 117:1. Complexation of siRNA prevented staining of siRNA by ethidium bromide and led to the disappearance of the siRNA band. Therefore, the fluorescent intensity of the siRNA band on the gel (well 5) disappeared when siRNA was complexed with NLC at w/w ratio = 117:1. (B) siRNA complexation efficiency of LHRH-NLC-TAX evaluated by the ethidium bromide dye displacement assay. The arrow on the curve highlights the w/w ratio corresponding to the apparent end of siRNA complexation with NLC. Means ± SD are shown.

Figure 4
Figure 4

Viability of multidrug-resistant H69AR human lung cancer cells incubated for 48 hours with the indicated formulations. A, cytotoxicity of formulations that do not contain DTAX; B, cytotoxicity of formulations that contain TAX. Means ± SD are shown. *P < 0.05 when compared with control.

Figure 5
Figure 5

Accumulation of LHRH-NLC in the lung tissues after the inhalation delivery (A) and intracellular localization of the drug and siRNA released from NLC (B-E). Representative transmission electron microscopy image of LHRH-targeted NLC labeled with osmium tetroxide in lung tissues (A). Representative images of A549 human lung cancer cells incubated 3 h with LHRH-NLC-DOX-siRNA: (B) visible light; (C) fluorescence images of nuclei stained with DAPI; (D) and (E) cellular localization of DOX (red fluorescence) and fluorescently labeled siRNA (green fluorescence), respectively.

Figure 6
Figure 6

Expression of MRP1 (A), BCL2 (B) and β2-m (C and D, internal standard) genes. Representative images of RT-PCR products and densitometric analysis of bands in A549 lung cancer cells incubated with the following formulations: (1) Control (medium); (2) LHRH-NLC-TAX; and (3) LHRH-NLC-TAX-siRNAs (MRP1 and BCL2). Gene expression in control was set to 100%. Means ± SD are shown. *P < 0.05 when compared with control.

Figure 7
Figure 7

Evaluation of orthotopic lung cancer model by imaging. (A) Bioluminescence optical imaging of control mouse and mice with lung tumors of different size. (B-D) Magnetic resonance imaging of control mouse (B) and mice with lung tumors of different size (C, D). Lung tumor (blue) and healthy lung tissues (red) are shown (D). (E) Optical imaging of excised organs. (F-G) Computed tomography images of control mouse (F) and mouse with lung tumors (G). (H) Visualization of lung tumor by ultrasound imaging system.

Figure 8
Figure 8

Accumulation of NLC in the lungs and other organs. (A) Organ distribution of fluorescently labeled (Cy5.5) NLC in mice after i.v. (left) and inhalation (right) administration. (B) Distribution of fluorescently labeled (Cy5.5) non-targeted and LHRH-tumor targeted NLC in mouse lungs bearing human lung cancer. (C) Distribution of fluorescently labeled (Cy5.5) LHRH-tumor - targeted (NLC-LHRH) in mouse lungs bearing human lung tumor cells (tumor and non-tumor tissues; bright field and fluorescence microscope images; red color represents distribution of NLC-LHRH in tumor and non-tumor lung tissues).

Figure 9
Figure 9

Changes in lung tumor volume after beginning of treatment. Mice were treated on days 0, 3, 7, 11, 14, 17, 21, and 24. 1 – Untreated mice; 2 – LHRH-NLC (inhalation); 3 – TAX (i.v.); 4 – LHRH-NLC-TAX (inhalation); 5 – LHRH-NLC-TAX-siRNAs targeted to MRP1 and BCL2 mRNAs (inhalation). Means ± SD are shown.

Similar articles

Cited by

References

    1. Carbone DP, Felip E. Adjuvant therapy in non-small cell lung cancer: future treatment prospects and paradigms. Clinical lung cancer. 2011;12:261–71. - PubMed
    1. Francis H, Solomon B. The current status of targeted therapy for non-small cell lung cancer. Internal medicine journal. 2010;40:611–8. - PubMed
    1. Higgins MJ, Ettinger DS. Chemotherapy for lung cancer: the state of the art in 2009. Expert review of anticancer therapy. 2009;9:1365–78. - PubMed
    1. Katzel JA, Fanucchi MP, Li Z. Recent advances of novel targeted therapy in non-small cell lung cancer. Journal of hematology & oncology. 2009;2:2. - PMC - PubMed
    1. Wagner TD, Yang GY. The role of chemotherapy and radiation in the treatment of locally advanced non-small cell lung cancer (NSCLC). Current drug targets. 2010;11:67–73. - PubMed

Publication types

MeSH terms

Substances