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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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In vivo tumor growth analysis and bioluminescence imaging of mice (n = 25) bearing MDA-MB-231 tumors stably expressing Fluc-eGFP that are treated with antisense-miR-21 and antisense-miR-10b loaded or coloaded with uPA–PLGA-b-PEG and Sc-uPA–PLGA-b-PEG NPs. (A) Tumors growth volume (mm3) measured in different treatment groups over time. (B) Optical bioluminescence images of animals (n = 10, 5 animals bearing two tumors each for each treatment group) treated with different NPs over time. (C) Quantitative graph showing the bioluminescence signals quantitated from animals shown in (B). (D) TUNEL staining of tumor tissues of animals treated with different NPs.
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fig5: In vivo tumor growth analysis and bioluminescence imaging of mice (n = 25) bearing MDA-MB-231 tumors stably expressing Fluc-eGFP that are treated with antisense-miR-21 and antisense-miR-10b loaded or coloaded with uPA–PLGA-b-PEG and Sc-uPA–PLGA-b-PEG NPs. (A) Tumors growth volume (mm3) measured in different treatment groups over time. (B) Optical bioluminescence images of animals (n = 10, 5 animals bearing two tumors each for each treatment group) treated with different NPs over time. (C) Quantitative graph showing the bioluminescence signals quantitated from animals shown in (B). (D) TUNEL staining of tumor tissues of animals treated with different NPs.

Mentions: The antitumor effect of systemically injected uPAR targeted NPs loaded with different combination of antisense-miRNAs was studied in tumor xenografts of MDA-MB-231 cells stably expressing Fluc-eGFP reporter fusion gene. The targeted delivery of NPs loaded with antisense-miR-21 and antisense-miR-10b, and coloaded with antisense-miR-21 and antisense-miR-10b combination, injected at 0.15 mg/kg, was measured for tumor size and bioluminescence imaging signal over time. The scrambled-uPA-peptide conjugated PLGA-b-PEG-NPs coloaded with antisense-miR-21 and antisense-miR-10b, and control PLGA-b-PEG-NPs injected animals served as controls. The mouse TNBC tumor xenograft model was developed by injecting 0.1 mL of MDA-MB-231-Fluc-eGFP cell suspension (10 × 106) into the left and right flanks of female nude mice (Nu/Nu) using 50% (v/v) Matrigel. After 15 days, when the tumors size reached 100–300 cubic mm, the animals [n = 25, 5 animals with two tumors each (n = 10 for each treatment group) for each treatment group] were intravenously injected with the NPs on Day 0, Day 3, and Day 6 (Figure 5A). The tumor development and growth was monitored over time (up to Day 9) by measuring tumor volume and bioluminescence imaging. After the completion of imaging on Day 9, the tumors were excised for ex vivo histological analysis. The results indicate that the tumor growth of animals treated by NPs loaded with both antisense-miR-10b and antisense-miR-21 individually and in combinations showed significant reduction in tumor growth, compared to control-NPs treated animals, with highest effect being detected in animals treated by uPA–PLGA-b-PEG-NPs coloaded with antisense-miR-10b and antisense-miR-21 combinations. The antisense-miR-21-10b coloaded-uPA NPs treated mice shows 40% more tumor reduction compared to antisense-miR-21-10b coloaded NPs with scrambled-uPA, signifying the uPA targeting effects on MDA-MB-231 tumors (Figure 5A). The bioluminescence imaging signal also showed similar level of signal drop in animals treated with antisense-miRNAs loaded NPs compared to control-NPs treated group (Figure 5A-C). The ex vivo analysis of kidney and liver tissues showed no sign of toxicity in animals treated by NPs with or without antisense-miRNAs (SI Figure S18A–C). The ex vivo TUNEL staining of tumor tissues showed significant amount of apoptotic cells in animals treated with antisense-miRNAs loaded NPs, with the highest level being detected in animals treated with uPA-targeted NPs coloaded with both antisense-miR-10b and antisense-miR-21 combination (Figure 5D, SI Figure S18D).


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)

In vivo tumor growth analysis and bioluminescence imaging of mice (n = 25) bearing MDA-MB-231 tumors stably expressing Fluc-eGFP that are treated with antisense-miR-21 and antisense-miR-10b loaded or coloaded with uPA–PLGA-b-PEG and Sc-uPA–PLGA-b-PEG NPs. (A) Tumors growth volume (mm3) measured in different treatment groups over time. (B) Optical bioluminescence images of animals (n = 10, 5 animals bearing two tumors each for each treatment group) treated with different NPs over time. (C) Quantitative graph showing the bioluminescence signals quantitated from animals shown in (B). (D) TUNEL staining of tumor tissues of animals treated with different NPs.
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fig5: In vivo tumor growth analysis and bioluminescence imaging of mice (n = 25) bearing MDA-MB-231 tumors stably expressing Fluc-eGFP that are treated with antisense-miR-21 and antisense-miR-10b loaded or coloaded with uPA–PLGA-b-PEG and Sc-uPA–PLGA-b-PEG NPs. (A) Tumors growth volume (mm3) measured in different treatment groups over time. (B) Optical bioluminescence images of animals (n = 10, 5 animals bearing two tumors each for each treatment group) treated with different NPs over time. (C) Quantitative graph showing the bioluminescence signals quantitated from animals shown in (B). (D) TUNEL staining of tumor tissues of animals treated with different NPs.
Mentions: The antitumor effect of systemically injected uPAR targeted NPs loaded with different combination of antisense-miRNAs was studied in tumor xenografts of MDA-MB-231 cells stably expressing Fluc-eGFP reporter fusion gene. The targeted delivery of NPs loaded with antisense-miR-21 and antisense-miR-10b, and coloaded with antisense-miR-21 and antisense-miR-10b combination, injected at 0.15 mg/kg, was measured for tumor size and bioluminescence imaging signal over time. The scrambled-uPA-peptide conjugated PLGA-b-PEG-NPs coloaded with antisense-miR-21 and antisense-miR-10b, and control PLGA-b-PEG-NPs injected animals served as controls. The mouse TNBC tumor xenograft model was developed by injecting 0.1 mL of MDA-MB-231-Fluc-eGFP cell suspension (10 × 106) into the left and right flanks of female nude mice (Nu/Nu) using 50% (v/v) Matrigel. After 15 days, when the tumors size reached 100–300 cubic mm, the animals [n = 25, 5 animals with two tumors each (n = 10 for each treatment group) for each treatment group] were intravenously injected with the NPs on Day 0, Day 3, and Day 6 (Figure 5A). The tumor development and growth was monitored over time (up to Day 9) by measuring tumor volume and bioluminescence imaging. After the completion of imaging on Day 9, the tumors were excised for ex vivo histological analysis. The results indicate that the tumor growth of animals treated by NPs loaded with both antisense-miR-10b and antisense-miR-21 individually and in combinations showed significant reduction in tumor growth, compared to control-NPs treated animals, with highest effect being detected in animals treated by uPA–PLGA-b-PEG-NPs coloaded with antisense-miR-10b and antisense-miR-21 combinations. The antisense-miR-21-10b coloaded-uPA NPs treated mice shows 40% more tumor reduction compared to antisense-miR-21-10b coloaded NPs with scrambled-uPA, signifying the uPA targeting effects on MDA-MB-231 tumors (Figure 5A). The bioluminescence imaging signal also showed similar level of signal drop in animals treated with antisense-miRNAs loaded NPs compared to control-NPs treated group (Figure 5A-C). The ex vivo analysis of kidney and liver tissues showed no sign of toxicity in animals treated by NPs with or without antisense-miRNAs (SI Figure S18A–C). The ex vivo TUNEL staining of tumor tissues showed significant amount of apoptotic cells in animals treated with antisense-miRNAs loaded NPs, with the highest level being detected in animals treated with uPA-targeted NPs coloaded with both antisense-miR-10b and antisense-miR-21 combination (Figure 5D, SI Figure S18D).

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