Cargando…

Active Tumor-Targeting Nano-formulations Containing Simvastatin and Doxorubicin Inhibit Melanoma Growth and Angiogenesis

Primary melanoma aggressiveness is determined by rapid selection and growth of cellular clones resistant to conventional treatments, resulting in metastasis and recurrence. In addition, a reprogrammed tumor-immune microenvironment supports melanoma progression and response to therapy. There is an ur...

Descripción completa

Detalles Bibliográficos
Autores principales: Negrea, Giorgiana, Rauca, Valentin-Florian, Meszaros, Marta Szilvia, Patras, Laura, Luput, Lavinia, Licarete, Emilia, Toma, Vlad-Alexandru, Porfire, Alina, Muntean, Dana, Sesarman, Alina, Banciu, Manuela
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9016200/
https://www.ncbi.nlm.nih.gov/pubmed/35450036
http://dx.doi.org/10.3389/fphar.2022.870347
_version_ 1784688478586404864
author Negrea, Giorgiana
Rauca, Valentin-Florian
Meszaros, Marta Szilvia
Patras, Laura
Luput, Lavinia
Licarete, Emilia
Toma, Vlad-Alexandru
Porfire, Alina
Muntean, Dana
Sesarman, Alina
Banciu, Manuela
author_facet Negrea, Giorgiana
Rauca, Valentin-Florian
Meszaros, Marta Szilvia
Patras, Laura
Luput, Lavinia
Licarete, Emilia
Toma, Vlad-Alexandru
Porfire, Alina
Muntean, Dana
Sesarman, Alina
Banciu, Manuela
author_sort Negrea, Giorgiana
collection PubMed
description Primary melanoma aggressiveness is determined by rapid selection and growth of cellular clones resistant to conventional treatments, resulting in metastasis and recurrence. In addition, a reprogrammed tumor-immune microenvironment supports melanoma progression and response to therapy. There is an urgent need to develop selective and specific drug delivery strategies for modulating the interaction between cancer cells and immune cells within the tumor microenvironment. This study proposes a novel combination therapy consisting of sequential administration of simvastatin incorporated in IL-13-functionalized long-circulating liposomes (IL-13-LCL-SIM) and doxorubicin encapsulated into PEG-coated extracellular vesicles (PEG-EV-DOX) to selectively target both tumor-associated macrophages and melanoma cells. To this end, IL-13 was conjugated to LCL-SIM which was obtained via the lipid film hydration method. EVs enriched from melanoma cells were passively loaded with doxorubicin. The cellular uptake of rhodamine-tagged nano-particles and the antiproliferative potential of the treatments by using the ELISA BrdU-colorimetric immunoassay were investigated in vitro. Subsequently, the therapeutic agents were administered i.v in B16.F10 melanoma-bearing mice, and tumor size was monitored during treatment. The molecular mechanisms of antitumor activity were investigated using angiogenic and inflammatory protein arrays and western blot analysis of invasion (HIF-1) and apoptosis markers (Bcl-xL and Bax). Quantification of oxidative stress marker malondialdehyde (MDA) was determined by HPLC. Immunohistochemical staining of angiogenic markers CD31 and VEGF and of pan-macrophage marker F4/80 was performed to validate our findings. The in vitro data showed that IL-13-functionalized LCL were preferentially taken up by tumor-associated macrophages and indicated that sequential administration of IL-13-LCL-SIM and PEG-EV-DOX had the strongest antiproliferative effect on tumor cells co-cultured with tumor-associated macrophages (TAMs). Accordingly, strong inhibition of tumor growth in the group treated with the sequential combination therapy was reported in vivo. Our data suggested that the antitumor action of the combined treatment was exerted through strong inhibition of several pro-angiogenic factors (VEGF, bFGF, and CD31) and oxidative stress-induced upregulation of pro-apoptotic protein Bax. This novel drug delivery strategy based on combined active targeting of both cancer cells and immune cells was able to induce a potent antitumor effect by disruption of the reciprocal interactions between TAMs and melanoma cells.
format Online
Article
Text
id pubmed-9016200
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-90162002022-04-20 Active Tumor-Targeting Nano-formulations Containing Simvastatin and Doxorubicin Inhibit Melanoma Growth and Angiogenesis Negrea, Giorgiana Rauca, Valentin-Florian Meszaros, Marta Szilvia Patras, Laura Luput, Lavinia Licarete, Emilia Toma, Vlad-Alexandru Porfire, Alina Muntean, Dana Sesarman, Alina Banciu, Manuela Front Pharmacol Pharmacology Primary melanoma aggressiveness is determined by rapid selection and growth of cellular clones resistant to conventional treatments, resulting in metastasis and recurrence. In addition, a reprogrammed tumor-immune microenvironment supports melanoma progression and response to therapy. There is an urgent need to develop selective and specific drug delivery strategies for modulating the interaction between cancer cells and immune cells within the tumor microenvironment. This study proposes a novel combination therapy consisting of sequential administration of simvastatin incorporated in IL-13-functionalized long-circulating liposomes (IL-13-LCL-SIM) and doxorubicin encapsulated into PEG-coated extracellular vesicles (PEG-EV-DOX) to selectively target both tumor-associated macrophages and melanoma cells. To this end, IL-13 was conjugated to LCL-SIM which was obtained via the lipid film hydration method. EVs enriched from melanoma cells were passively loaded with doxorubicin. The cellular uptake of rhodamine-tagged nano-particles and the antiproliferative potential of the treatments by using the ELISA BrdU-colorimetric immunoassay were investigated in vitro. Subsequently, the therapeutic agents were administered i.v in B16.F10 melanoma-bearing mice, and tumor size was monitored during treatment. The molecular mechanisms of antitumor activity were investigated using angiogenic and inflammatory protein arrays and western blot analysis of invasion (HIF-1) and apoptosis markers (Bcl-xL and Bax). Quantification of oxidative stress marker malondialdehyde (MDA) was determined by HPLC. Immunohistochemical staining of angiogenic markers CD31 and VEGF and of pan-macrophage marker F4/80 was performed to validate our findings. The in vitro data showed that IL-13-functionalized LCL were preferentially taken up by tumor-associated macrophages and indicated that sequential administration of IL-13-LCL-SIM and PEG-EV-DOX had the strongest antiproliferative effect on tumor cells co-cultured with tumor-associated macrophages (TAMs). Accordingly, strong inhibition of tumor growth in the group treated with the sequential combination therapy was reported in vivo. Our data suggested that the antitumor action of the combined treatment was exerted through strong inhibition of several pro-angiogenic factors (VEGF, bFGF, and CD31) and oxidative stress-induced upregulation of pro-apoptotic protein Bax. This novel drug delivery strategy based on combined active targeting of both cancer cells and immune cells was able to induce a potent antitumor effect by disruption of the reciprocal interactions between TAMs and melanoma cells. Frontiers Media S.A. 2022-04-05 /pmc/articles/PMC9016200/ /pubmed/35450036 http://dx.doi.org/10.3389/fphar.2022.870347 Text en Copyright © 2022 Negrea, Rauca, Meszaros, Patras, Luput, Licarete, Toma, Porfire, Muntean, Sesarman and Banciu. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Pharmacology
Negrea, Giorgiana
Rauca, Valentin-Florian
Meszaros, Marta Szilvia
Patras, Laura
Luput, Lavinia
Licarete, Emilia
Toma, Vlad-Alexandru
Porfire, Alina
Muntean, Dana
Sesarman, Alina
Banciu, Manuela
Active Tumor-Targeting Nano-formulations Containing Simvastatin and Doxorubicin Inhibit Melanoma Growth and Angiogenesis
title Active Tumor-Targeting Nano-formulations Containing Simvastatin and Doxorubicin Inhibit Melanoma Growth and Angiogenesis
title_full Active Tumor-Targeting Nano-formulations Containing Simvastatin and Doxorubicin Inhibit Melanoma Growth and Angiogenesis
title_fullStr Active Tumor-Targeting Nano-formulations Containing Simvastatin and Doxorubicin Inhibit Melanoma Growth and Angiogenesis
title_full_unstemmed Active Tumor-Targeting Nano-formulations Containing Simvastatin and Doxorubicin Inhibit Melanoma Growth and Angiogenesis
title_short Active Tumor-Targeting Nano-formulations Containing Simvastatin and Doxorubicin Inhibit Melanoma Growth and Angiogenesis
title_sort active tumor-targeting nano-formulations containing simvastatin and doxorubicin inhibit melanoma growth and angiogenesis
topic Pharmacology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9016200/
https://www.ncbi.nlm.nih.gov/pubmed/35450036
http://dx.doi.org/10.3389/fphar.2022.870347
work_keys_str_mv AT negreagiorgiana activetumortargetingnanoformulationscontainingsimvastatinanddoxorubicininhibitmelanomagrowthandangiogenesis
AT raucavalentinflorian activetumortargetingnanoformulationscontainingsimvastatinanddoxorubicininhibitmelanomagrowthandangiogenesis
AT meszarosmartaszilvia activetumortargetingnanoformulationscontainingsimvastatinanddoxorubicininhibitmelanomagrowthandangiogenesis
AT patraslaura activetumortargetingnanoformulationscontainingsimvastatinanddoxorubicininhibitmelanomagrowthandangiogenesis
AT luputlavinia activetumortargetingnanoformulationscontainingsimvastatinanddoxorubicininhibitmelanomagrowthandangiogenesis
AT licareteemilia activetumortargetingnanoformulationscontainingsimvastatinanddoxorubicininhibitmelanomagrowthandangiogenesis
AT tomavladalexandru activetumortargetingnanoformulationscontainingsimvastatinanddoxorubicininhibitmelanomagrowthandangiogenesis
AT porfirealina activetumortargetingnanoformulationscontainingsimvastatinanddoxorubicininhibitmelanomagrowthandangiogenesis
AT munteandana activetumortargetingnanoformulationscontainingsimvastatinanddoxorubicininhibitmelanomagrowthandangiogenesis
AT sesarmanalina activetumortargetingnanoformulationscontainingsimvastatinanddoxorubicininhibitmelanomagrowthandangiogenesis
AT banciumanuela activetumortargetingnanoformulationscontainingsimvastatinanddoxorubicininhibitmelanomagrowthandangiogenesis