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Coating Solid Lipid Nanoparticles with Hyaluronic Acid Enhances Antitumor Activity against Melanoma Stem-like Cells.

Shen H, Shi S, Zhang Z, Gong T, Sun X - Theranostics (2015)

Bottom Line: First, we developed a model system based on melanoma stem-like cells for experiments in vitro and in mouse xenografts, and we showed that cells expressing high levels of CD44 (CD44(+)) displayed a strong CSC phenotype while cells expressing low levels of CD44 (CD44(-)) did not.In the B16F10-CD44(+) lung metastasis model, PTX-loaded HA-SLNs targeted the tumor-bearing lung tissues well and subsequently exhibited significant antitumor effects with a relative low dose of PTX, which provided significant survival benefit without evidence of adverse events.These findings suggest that the HA-SLNs targeting system shows promise for enhancing cancer therapy.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Drug Targeting and Drug Delivery Systems, Ministry of Education, West China School of Pharmacy, Sichuan University, Chengdu 610041, P. R. China.

ABSTRACT
Successful anticancer chemotherapy requires targeting tumors efficiently and further potential to eliminate cancer stem cell (CSC) subpopulations. Since CD44 is present on many types of CSCs, and it binds specially to hyaluronic acid (HA), we tested whether coating solid lipid nanoparticles with hyaluronan (HA-SLNs)would allow targeted delivery of paclitaxel (PTX) to CD44-overexpressing B16F10 melanoma cells. First, we developed a model system based on melanoma stem-like cells for experiments in vitro and in mouse xenografts, and we showed that cells expressing high levels of CD44 (CD44(+)) displayed a strong CSC phenotype while cells expressing low levels of CD44 (CD44(-)) did not. This phenotype included sphere and colony formation, higher proportion of side population cells, expression of CSC-related markers (ALDH, CD133, Oct-4) and tumorigenicity in vivo. Next we showed that administering PTX-loaded HA-SLNs led to efficient intracellular delivery of PTX and induced substantial apoptosis in CD44(+) cells in vitro. In the B16F10-CD44(+) lung metastasis model, PTX-loaded HA-SLNs targeted the tumor-bearing lung tissues well and subsequently exhibited significant antitumor effects with a relative low dose of PTX, which provided significant survival benefit without evidence of adverse events. These findings suggest that the HA-SLNs targeting system shows promise for enhancing cancer therapy.

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A) Flow cytometry analysis to measure the efficiency of cellular uptake of uncoated SLNs/PTX, heparin-coated SLNs/PTX and HA-SLNs/PTX (Coumarin-6 was incorporated in the place of PTX as a fluorescent marker). Each preparation was incubated with B16F10-CD44+ cells for 1 h. B) Efficiency of cell binding by HA-SLNs/PTX in the presence of excess free HA or anti-CD44 antibody (n = 3). C) Cell-binding ability of HA-SLNs/PTX (loading coumarin-6) in CD44 high expression- and low expression-cells. Data shown were mean ± SD (n = 3). D) Confocal laser scanning microscopy of uptake of HA-SLNs/PTX (loading coumarin-6) by B16F10-CD44+ cells at 37°C in the presence of excess free HA or anti-CD44 antibody. Scale bar, 25 μm. *p < 0.05, **p < 0.01, compared with cells treated with HA-SLNs/PTX.
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Figure 3: A) Flow cytometry analysis to measure the efficiency of cellular uptake of uncoated SLNs/PTX, heparin-coated SLNs/PTX and HA-SLNs/PTX (Coumarin-6 was incorporated in the place of PTX as a fluorescent marker). Each preparation was incubated with B16F10-CD44+ cells for 1 h. B) Efficiency of cell binding by HA-SLNs/PTX in the presence of excess free HA or anti-CD44 antibody (n = 3). C) Cell-binding ability of HA-SLNs/PTX (loading coumarin-6) in CD44 high expression- and low expression-cells. Data shown were mean ± SD (n = 3). D) Confocal laser scanning microscopy of uptake of HA-SLNs/PTX (loading coumarin-6) by B16F10-CD44+ cells at 37°C in the presence of excess free HA or anti-CD44 antibody. Scale bar, 25 μm. *p < 0.05, **p < 0.01, compared with cells treated with HA-SLNs/PTX.

Mentions: B16F10-CD44+ cells took up HA-SLNs/PTX to a much greater extent than the nanoparticles lacking HA or the same nanoparticles coated with the similar mucopolysaccharide heparin, even though the heparin-coated vesicles showed a similar zeta potential (32.3 ± 1.56 mV) and particle size (183.3 ± 1.2192 nm). This suggested that uptake occurred through a specific, HA-mediated mechanism. Experiments in Fig. 3A and Supplementary Material: Fig. S1 suggested that the high affinity of HA-SLNs/PTX to the B16F10-CD44+ cells was not only attributed to nonspecific electrostatic attraction between the nanoparticles and the cell surface, but also because of specific HA binding to CD44.


Coating Solid Lipid Nanoparticles with Hyaluronic Acid Enhances Antitumor Activity against Melanoma Stem-like Cells.

Shen H, Shi S, Zhang Z, Gong T, Sun X - Theranostics (2015)

A) Flow cytometry analysis to measure the efficiency of cellular uptake of uncoated SLNs/PTX, heparin-coated SLNs/PTX and HA-SLNs/PTX (Coumarin-6 was incorporated in the place of PTX as a fluorescent marker). Each preparation was incubated with B16F10-CD44+ cells for 1 h. B) Efficiency of cell binding by HA-SLNs/PTX in the presence of excess free HA or anti-CD44 antibody (n = 3). C) Cell-binding ability of HA-SLNs/PTX (loading coumarin-6) in CD44 high expression- and low expression-cells. Data shown were mean ± SD (n = 3). D) Confocal laser scanning microscopy of uptake of HA-SLNs/PTX (loading coumarin-6) by B16F10-CD44+ cells at 37°C in the presence of excess free HA or anti-CD44 antibody. Scale bar, 25 μm. *p < 0.05, **p < 0.01, compared with cells treated with HA-SLNs/PTX.
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Figure 3: A) Flow cytometry analysis to measure the efficiency of cellular uptake of uncoated SLNs/PTX, heparin-coated SLNs/PTX and HA-SLNs/PTX (Coumarin-6 was incorporated in the place of PTX as a fluorescent marker). Each preparation was incubated with B16F10-CD44+ cells for 1 h. B) Efficiency of cell binding by HA-SLNs/PTX in the presence of excess free HA or anti-CD44 antibody (n = 3). C) Cell-binding ability of HA-SLNs/PTX (loading coumarin-6) in CD44 high expression- and low expression-cells. Data shown were mean ± SD (n = 3). D) Confocal laser scanning microscopy of uptake of HA-SLNs/PTX (loading coumarin-6) by B16F10-CD44+ cells at 37°C in the presence of excess free HA or anti-CD44 antibody. Scale bar, 25 μm. *p < 0.05, **p < 0.01, compared with cells treated with HA-SLNs/PTX.
Mentions: B16F10-CD44+ cells took up HA-SLNs/PTX to a much greater extent than the nanoparticles lacking HA or the same nanoparticles coated with the similar mucopolysaccharide heparin, even though the heparin-coated vesicles showed a similar zeta potential (32.3 ± 1.56 mV) and particle size (183.3 ± 1.2192 nm). This suggested that uptake occurred through a specific, HA-mediated mechanism. Experiments in Fig. 3A and Supplementary Material: Fig. S1 suggested that the high affinity of HA-SLNs/PTX to the B16F10-CD44+ cells was not only attributed to nonspecific electrostatic attraction between the nanoparticles and the cell surface, but also because of specific HA binding to CD44.

Bottom Line: First, we developed a model system based on melanoma stem-like cells for experiments in vitro and in mouse xenografts, and we showed that cells expressing high levels of CD44 (CD44(+)) displayed a strong CSC phenotype while cells expressing low levels of CD44 (CD44(-)) did not.In the B16F10-CD44(+) lung metastasis model, PTX-loaded HA-SLNs targeted the tumor-bearing lung tissues well and subsequently exhibited significant antitumor effects with a relative low dose of PTX, which provided significant survival benefit without evidence of adverse events.These findings suggest that the HA-SLNs targeting system shows promise for enhancing cancer therapy.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Drug Targeting and Drug Delivery Systems, Ministry of Education, West China School of Pharmacy, Sichuan University, Chengdu 610041, P. R. China.

ABSTRACT
Successful anticancer chemotherapy requires targeting tumors efficiently and further potential to eliminate cancer stem cell (CSC) subpopulations. Since CD44 is present on many types of CSCs, and it binds specially to hyaluronic acid (HA), we tested whether coating solid lipid nanoparticles with hyaluronan (HA-SLNs)would allow targeted delivery of paclitaxel (PTX) to CD44-overexpressing B16F10 melanoma cells. First, we developed a model system based on melanoma stem-like cells for experiments in vitro and in mouse xenografts, and we showed that cells expressing high levels of CD44 (CD44(+)) displayed a strong CSC phenotype while cells expressing low levels of CD44 (CD44(-)) did not. This phenotype included sphere and colony formation, higher proportion of side population cells, expression of CSC-related markers (ALDH, CD133, Oct-4) and tumorigenicity in vivo. Next we showed that administering PTX-loaded HA-SLNs led to efficient intracellular delivery of PTX and induced substantial apoptosis in CD44(+) cells in vitro. In the B16F10-CD44(+) lung metastasis model, PTX-loaded HA-SLNs targeted the tumor-bearing lung tissues well and subsequently exhibited significant antitumor effects with a relative low dose of PTX, which provided significant survival benefit without evidence of adverse events. These findings suggest that the HA-SLNs targeting system shows promise for enhancing cancer therapy.

Show MeSH
Related in: MedlinePlus