Targeted combination therapy for glioblastoma by co-delivery of doxorubicin, YAP-siRNA and gold nanorods

doi:10.1016/j.jmst.2020.03.009

Abstract

Abstract:

The combination of brain targeting drug delivery systems and multi-modal intervention pose a promising therapeutic approach for glioblastoma therapy. In this study, we developed an angiopep-2 peptide modified cationic liposome loaded with doxorubicin, YAP-siRNA and gold nanorods (D/R@Ang2-Lip + Au) simultaneously, which has high encapsulating efficiency for doxorubicin (95.4 %) and effective binding of siRNA at N/P ratio of 20:1. The fluorescence imaging and flow cytometry analysis revealed high cellular uptake of D/R@Ang2-Lip + Au. Real-time quantitative polymerase chain reaction and western blot analysis indicated that D/R@Ang2-Lip + Au could effectively inhibit the expression of YAP protein. In vitro and in vivo studies showed that D/R@Ang2-Lip + Au had the ability to target glioblastoma cells, and achieved better anti-proliferative effects compared with non-targeted D/R@Lip + Au. Moreover, in vivo experiment demonstrated that D/R@Ang2-Lip + Au was able to cross the blood-brain barrier, and combination therapy could significantly inhibit tumor growth. Therefore, the multifunctional D/R@Ang2-Lip + Au might provide a novel approach for effectively delivery of DOX, YAP-siRNA and AuNRs into the glioblastoma cells simultaneously and exerting synergistic therapeutic effects.

Key words: Cationic liposomes, Angiopep-2, Doxorubicin, YAP siRNA, AuNRs, Glioblastoma treatment

Lihuang Li, Qiuyan Guo, Yanxiu Liu, Mindan Lu, Jun Yang, Yunlong Ge, Qiang Zhang, Benqiang Sun, Xiumin Wang, Liang-cheng Li, Lei Ren. Targeted combination therapy for glioblastoma by co-delivery of doxorubicin, YAP-siRNA and gold nanorods[J]. J. Mater. Sci. Technol., 2021, 63: 81-90.

Figures/Tables 7

Table 1 Nomenclature and feature of liposome formulations.

Nomenclature	Feature
Ang2-Lip	liposomes with angiopep-2
DOX@Lip	liposomes loaded with DOX
RNA@Lip	liposomes loaded with YAP-siRNA
D/R@Lip	liposomes loaded with DOX and YAP-siRNA
Lip + Au	liposomes complexed with gold nanorods

Fig. 1. Schematic of the preparation process of D/R@Ang2-Lip + Au, and the anti- glioblastoma mechanisms.

Fig. 2. (A) Typical TEM images of D/R@Ang2-Lip + Au. (B) The size and zeta potential of different liposome formulations. (C) The UV-vis absorbance spectra of Lip, AuNRs, Ang2-Lip + Au, and D/R@Ang2-Lip + Au. (D) In vitro release profiles of free DOX and D/R@Ang2-Lip + Au with or without 808 nm NIR laser irradiation. (E) Agarose gel electrophoresis retardation assay for D/R@Ang2-Lip + Au with 1 % DSPE-PEG2000-Ang2. (F) Temperature change curves of D/R@Ang2-Lip + Au solutions with different concentration of AuNRs under 808 nm NIR laser irradiation (1 W/cm2, 5 min).

Fig. 3. CLSM images of U87MG cells incubated with PBS, XsTR/siRNA, free DOX, D/R@Lip + Au, and D/R@Ang2-Lip + Au.

Fig. 4. Flow cytometry of cell uptake of (A) DOX and (B) siRNA in U87MG cells. (C) mRNA levels of YAP in U87MG cells infected with various formulations were examined via qPCR. (D) Western blotting of YAP levels in U87 MG cells infected with various formulations. Data are expressed as mean ± SD (n = 3). * p < 0.05; ** p < 0.01.

Fig. 5. (A) Relative cell viability of U87MG cells incubated with targeting liposomes formulations. (B) Quantitative analysis of siRNA distribution in various excised organs. (C) The fluorescence images treated with different formulations. (D) Body weight change of mice treated with different formulations. Data are expressed as mean ± SD (n = 3). * p < 0.05; ** p < 0.01.

Fig. 6. (A) H&E stained images of heart, liver, spleen, lung, kidney and brain collected from mice of all the experimental groups. (B) Histological images of the glioblastoma tissues using TUNEL assay. Magnification: 200 × .

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