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Polymer nanoparticles mediated codelivery of antimiR-10b and antimiR-21 for achieving triple negative breast cancer therapy.

Devulapally R, Sekar NM, Sekar TV, Foygel K, Massoud TF, Willmann JK, Paulmurugan R - ACS Nano (2015)

Bottom Line: The current study shows the therapeutic outcome achieved in triple negative breast cancer (TNBC) by simultaneously antagonizing miR-21-induced antiapoptosis and miR-10b-induced metastasis, using antisense-miR-21-PS and antisense-miR-10b-PS delivered by polymer nanoparticles (NPs).We synthesized the antisense-miR-21 and antisense-miR-10b loaded PLGA-b-PEG polymer NPs and evaluated their cellular uptake, serum stability, release profile, and the subsequent synchronous blocking of endogenous miR-21 and miR-10b function in TNBC cells in culture, and tumor xenografts in living animals using molecular imaging.Targeted delivery of antisense-miR-21 and antisense-miR-10b coloaded urokinase plasminogen activator receptor (uPAR) targeted polymer NPs treated mice showed substantial reduction in tumor growth at very low dose of 0.15 mg/kg, compared to the control NPs treated mice and 40% reduction in tumor growth compared to scramble peptide conjugated NPs treated mice, thus demonstrating a potential new therapeutic option for TNBC.

View Article: PubMed Central - PubMed

Affiliation: Molecular Imaging Program at Stanford, Bio-X Program, Department of Radiology, Stanford University School of Medicine, Stanford University, 3155 Porter Drive, Palo Alto, California 94304, United States.

ABSTRACT
The current study shows the therapeutic outcome achieved in triple negative breast cancer (TNBC) by simultaneously antagonizing miR-21-induced antiapoptosis and miR-10b-induced metastasis, using antisense-miR-21-PS and antisense-miR-10b-PS delivered by polymer nanoparticles (NPs). We synthesized the antisense-miR-21 and antisense-miR-10b loaded PLGA-b-PEG polymer NPs and evaluated their cellular uptake, serum stability, release profile, and the subsequent synchronous blocking of endogenous miR-21 and miR-10b function in TNBC cells in culture, and tumor xenografts in living animals using molecular imaging. Results show that multitarget antagonization of endogenous miRNAs could be an efficient strategy for targeting metastasis and antiapoptosis in the treatment of metastatic cancer. Targeted delivery of antisense-miR-21 and antisense-miR-10b coloaded urokinase plasminogen activator receptor (uPAR) targeted polymer NPs treated mice showed substantial reduction in tumor growth at very low dose of 0.15 mg/kg, compared to the control NPs treated mice and 40% reduction in tumor growth compared to scramble peptide conjugated NPs treated mice, thus demonstrating a potential new therapeutic option for TNBC.

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Cytotoxicity evaluation of various antisense-miRNAs loaded PLGA-b-PEG-NPs in MDA-MB-231-Fluc-eGFP cells by MTT assay. (A–D) Cells treated with 0 to 25 pmol/mL miRNA equivalent of control NPs, antisense-miR-21 and antisense-miR-10b individually-and coloaded NPs for 24 h and assessed for cytotoxicity by MTT assay. (E–H) Cells treated with 12.5 and 25 pmol/mL miRNA equivalent of control NPs, antisense-miR-21 and antisense-miR-10b individually-and coloaded NPs for various time points (24–72 h) and assessed for cytotoxicity by MTT assay. Error bars are SEM of three determinants (*p < 0.05).
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fig3: Cytotoxicity evaluation of various antisense-miRNAs loaded PLGA-b-PEG-NPs in MDA-MB-231-Fluc-eGFP cells by MTT assay. (A–D) Cells treated with 0 to 25 pmol/mL miRNA equivalent of control NPs, antisense-miR-21 and antisense-miR-10b individually-and coloaded NPs for 24 h and assessed for cytotoxicity by MTT assay. (E–H) Cells treated with 12.5 and 25 pmol/mL miRNA equivalent of control NPs, antisense-miR-21 and antisense-miR-10b individually-and coloaded NPs for various time points (24–72 h) and assessed for cytotoxicity by MTT assay. Error bars are SEM of three determinants (*p < 0.05).

Mentions: PLGA-b-PEG NPs loaded with and without antisense-miRNAs were evaluated for induced cytotoxicity in MDA-MB-231 TNBC cells by MTT assay. The concentration dependent toxicity analysis in MDA-MB-231 cells shows that polymer equivalent of up to 50 μg/mL of PLGA-b-PEG NPs in media could be used without significant toxic effect. Hence, in all other antisense-miRNA therapeutic studies, NPs at PLGA-b-PEG polymer concentrations below 50 μg/mL was used. The MDA-MB-231 cells treated with 0–25 pmols of antisense-miRNA loaded NPs were evaluated for cell viability 24 h after treatment. The results show no toxicity in cells treated with control NPs. In contrast, the cells treated with antisense-miRNA loaded NPs show dose dependent reduction in cell viability, with the highest one resulting from coloaded-NPs with antisense-miRNAs concentrations (antisense-miR-21 and antisense-miR-10b) of 25 pmols each (17 ± 2% less viability compared to control). The cells treated with 25 pmols of antisense-miR-21 and antisense-miR-10b loaded NPs show 15 ± 2% and 13 ± 1% reduction in cell viability, respectively (Figure 3A–D). Time dependent cell viability analysis indicate that there was significant growth reduction without much cell death (assessed by PI staining-FACS) observed in cells exposed to antisense-miR-21 loaded NPs, and antisense-miR-21 and antisense-miR-10b coloaded NPs, as compared to cells treated with control NPs or antisense-miR-10b loaded NPs (Figure 3E–H). We also compared the effect of antisense-miRNAs delivered by PLGA-b-PEG-NPs with liposome-mediated transfection in cells. The results indicate significant cytotoxicity (P < 0.05) in MDA-MB-231 and MCF7 cells when cells were transfected with a combination of antisense-miRNA-21 and antisense-miRNA-10b by liposome (SI Figure S9) compared to cells treated with PLGA-b-PEG-NPs coloaded with a combination of antisense-miR-21 and antisense-miR-10b (Figure 3A–H). Moreover, a significant reduction in cell viability was observed in cells cotransfected with antisense-miRNA-21 and antisense-miRNA-10b combination by liposome compared to control cells (P = 0.0158 [antisense-miR-21:28 ± 3% reduction in cell viability] vs 0.0108 [antisense-miR-21 + antisense-miR-10b: 32 ± 2% reduction in cell viability]).


Polymer nanoparticles mediated codelivery of antimiR-10b and antimiR-21 for achieving triple negative breast cancer therapy.

Devulapally R, Sekar NM, Sekar TV, Foygel K, Massoud TF, Willmann JK, Paulmurugan R - ACS Nano (2015)

Cytotoxicity evaluation of various antisense-miRNAs loaded PLGA-b-PEG-NPs in MDA-MB-231-Fluc-eGFP cells by MTT assay. (A–D) Cells treated with 0 to 25 pmol/mL miRNA equivalent of control NPs, antisense-miR-21 and antisense-miR-10b individually-and coloaded NPs for 24 h and assessed for cytotoxicity by MTT assay. (E–H) Cells treated with 12.5 and 25 pmol/mL miRNA equivalent of control NPs, antisense-miR-21 and antisense-miR-10b individually-and coloaded NPs for various time points (24–72 h) and assessed for cytotoxicity by MTT assay. Error bars are SEM of three determinants (*p < 0.05).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4374409&req=5

fig3: Cytotoxicity evaluation of various antisense-miRNAs loaded PLGA-b-PEG-NPs in MDA-MB-231-Fluc-eGFP cells by MTT assay. (A–D) Cells treated with 0 to 25 pmol/mL miRNA equivalent of control NPs, antisense-miR-21 and antisense-miR-10b individually-and coloaded NPs for 24 h and assessed for cytotoxicity by MTT assay. (E–H) Cells treated with 12.5 and 25 pmol/mL miRNA equivalent of control NPs, antisense-miR-21 and antisense-miR-10b individually-and coloaded NPs for various time points (24–72 h) and assessed for cytotoxicity by MTT assay. Error bars are SEM of three determinants (*p < 0.05).
Mentions: PLGA-b-PEG NPs loaded with and without antisense-miRNAs were evaluated for induced cytotoxicity in MDA-MB-231 TNBC cells by MTT assay. The concentration dependent toxicity analysis in MDA-MB-231 cells shows that polymer equivalent of up to 50 μg/mL of PLGA-b-PEG NPs in media could be used without significant toxic effect. Hence, in all other antisense-miRNA therapeutic studies, NPs at PLGA-b-PEG polymer concentrations below 50 μg/mL was used. The MDA-MB-231 cells treated with 0–25 pmols of antisense-miRNA loaded NPs were evaluated for cell viability 24 h after treatment. The results show no toxicity in cells treated with control NPs. In contrast, the cells treated with antisense-miRNA loaded NPs show dose dependent reduction in cell viability, with the highest one resulting from coloaded-NPs with antisense-miRNAs concentrations (antisense-miR-21 and antisense-miR-10b) of 25 pmols each (17 ± 2% less viability compared to control). The cells treated with 25 pmols of antisense-miR-21 and antisense-miR-10b loaded NPs show 15 ± 2% and 13 ± 1% reduction in cell viability, respectively (Figure 3A–D). Time dependent cell viability analysis indicate that there was significant growth reduction without much cell death (assessed by PI staining-FACS) observed in cells exposed to antisense-miR-21 loaded NPs, and antisense-miR-21 and antisense-miR-10b coloaded NPs, as compared to cells treated with control NPs or antisense-miR-10b loaded NPs (Figure 3E–H). We also compared the effect of antisense-miRNAs delivered by PLGA-b-PEG-NPs with liposome-mediated transfection in cells. The results indicate significant cytotoxicity (P < 0.05) in MDA-MB-231 and MCF7 cells when cells were transfected with a combination of antisense-miRNA-21 and antisense-miRNA-10b by liposome (SI Figure S9) compared to cells treated with PLGA-b-PEG-NPs coloaded with a combination of antisense-miR-21 and antisense-miR-10b (Figure 3A–H). Moreover, a significant reduction in cell viability was observed in cells cotransfected with antisense-miRNA-21 and antisense-miRNA-10b combination by liposome compared to control cells (P = 0.0158 [antisense-miR-21:28 ± 3% reduction in cell viability] vs 0.0108 [antisense-miR-21 + antisense-miR-10b: 32 ± 2% reduction in cell viability]).

Bottom Line: The current study shows the therapeutic outcome achieved in triple negative breast cancer (TNBC) by simultaneously antagonizing miR-21-induced antiapoptosis and miR-10b-induced metastasis, using antisense-miR-21-PS and antisense-miR-10b-PS delivered by polymer nanoparticles (NPs).We synthesized the antisense-miR-21 and antisense-miR-10b loaded PLGA-b-PEG polymer NPs and evaluated their cellular uptake, serum stability, release profile, and the subsequent synchronous blocking of endogenous miR-21 and miR-10b function in TNBC cells in culture, and tumor xenografts in living animals using molecular imaging.Targeted delivery of antisense-miR-21 and antisense-miR-10b coloaded urokinase plasminogen activator receptor (uPAR) targeted polymer NPs treated mice showed substantial reduction in tumor growth at very low dose of 0.15 mg/kg, compared to the control NPs treated mice and 40% reduction in tumor growth compared to scramble peptide conjugated NPs treated mice, thus demonstrating a potential new therapeutic option for TNBC.

View Article: PubMed Central - PubMed

Affiliation: Molecular Imaging Program at Stanford, Bio-X Program, Department of Radiology, Stanford University School of Medicine, Stanford University, 3155 Porter Drive, Palo Alto, California 94304, United States.

ABSTRACT
The current study shows the therapeutic outcome achieved in triple negative breast cancer (TNBC) by simultaneously antagonizing miR-21-induced antiapoptosis and miR-10b-induced metastasis, using antisense-miR-21-PS and antisense-miR-10b-PS delivered by polymer nanoparticles (NPs). We synthesized the antisense-miR-21 and antisense-miR-10b loaded PLGA-b-PEG polymer NPs and evaluated their cellular uptake, serum stability, release profile, and the subsequent synchronous blocking of endogenous miR-21 and miR-10b function in TNBC cells in culture, and tumor xenografts in living animals using molecular imaging. Results show that multitarget antagonization of endogenous miRNAs could be an efficient strategy for targeting metastasis and antiapoptosis in the treatment of metastatic cancer. Targeted delivery of antisense-miR-21 and antisense-miR-10b coloaded urokinase plasminogen activator receptor (uPAR) targeted polymer NPs treated mice showed substantial reduction in tumor growth at very low dose of 0.15 mg/kg, compared to the control NPs treated mice and 40% reduction in tumor growth compared to scramble peptide conjugated NPs treated mice, thus demonstrating a potential new therapeutic option for TNBC.

Show MeSH
Related in: MedlinePlus