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1. Engineering pH-Sensitive Stable Nanovesicles for Delivery of MicroRNA Therapeutics

Author

Ministerio de Educación, Cultura y Deporte (España), Ministerio de Economía y Competitividad (España), Ministerio de Ciencia, Innovación y Universidades (España), Instituto de Salud Carlos III, Centro de Investigación Biomédica en Red Bioingeniería, Biomateriales y Nanomedicina (España), Matem lo Bitxo, Asociación Española Contra el Cáncer, Generalitat de Catalunya, Boloix, Ariadna [0000-0002-1648-5589], Köber, Mariana [0000-0001-9962-7900], Soriano, Aroa [0000-0001-9659-1471], Masanas, Marc [0000-0002-2249-8554], Segovia, Nathaly [0000-0001-8814-6095], Vargas Nadal, Guillem [0000-0003-4383-1325], Merlo Mas, Josep [0000-0002-3698-6655], Danino, Dganit [0000-0002-9782-4940], Foradada, Laia [0000-0002-0589-4360], Roma, Josep [0000-0001-7692-6123], Toledo, Josep Sánchez de [0000-0002-1034-1920], Gallego, Soledad [0000-0002-4712-9624], Veciana, Jaume [0000-0003-1023-9923], Albertazzi, Lorenzo [0000-0002-6837-0812], Segura, Miguel F. [0000-0003-0916-3618], Ventosa, Nora [0000-0002-8008-4974], Boloix, Ariadna, Feiner-Gracia, Natalia, Köber, Mariana, Repetto, Javier, Pascarella, Rosa, Soriano, Aroa, Masanas, Marc, Segovia, Nathaly, Vargas Nadal, Guillem, Merlo Mas, Josep, Danino, Dganit, Abutbul-Ionita, Inbal, Foradada, Laia, Roma, Josep, Córdoba, Alba, Sala Vergés, Santiago, Toledo, Josep Sánchez de, Gallego, Soledad, Veciana, Jaume, Albertazzi, Lorenzo, Segura, Miguel F., Ventosa, Nora, Ministerio de Educación, Cultura y Deporte (España), Ministerio de Economía y Competitividad (España), Ministerio de Ciencia, Innovación y Universidades (España), Instituto de Salud Carlos III, Centro de Investigación Biomédica en Red Bioingeniería, Biomateriales y Nanomedicina (España), Matem lo Bitxo, Asociación Española Contra el Cáncer, Generalitat de Catalunya, Boloix, Ariadna [0000-0002-1648-5589], Köber, Mariana [0000-0001-9962-7900], Soriano, Aroa [0000-0001-9659-1471], Masanas, Marc [0000-0002-2249-8554], Segovia, Nathaly [0000-0001-8814-6095], Vargas Nadal, Guillem [0000-0003-4383-1325], Merlo Mas, Josep [0000-0002-3698-6655], Danino, Dganit [0000-0002-9782-4940], Foradada, Laia [0000-0002-0589-4360], Roma, Josep [0000-0001-7692-6123], Toledo, Josep Sánchez de [0000-0002-1034-1920], Gallego, Soledad [0000-0002-4712-9624], Veciana, Jaume [0000-0003-1023-9923], Albertazzi, Lorenzo [0000-0002-6837-0812], Segura, Miguel F. [0000-0003-0916-3618], Ventosa, Nora [0000-0002-8008-4974], Boloix, Ariadna, Feiner-Gracia, Natalia, Köber, Mariana, Repetto, Javier, Pascarella, Rosa, Soriano, Aroa, Masanas, Marc, Segovia, Nathaly, Vargas Nadal, Guillem, Merlo Mas, Josep, Danino, Dganit, Abutbul-Ionita, Inbal, Foradada, Laia, Roma, Josep, Córdoba, Alba, Sala Vergés, Santiago, Toledo, Josep Sánchez de, Gallego, Soledad, Veciana, Jaume, Albertazzi, Lorenzo, Segura, Miguel F., and Ventosa, Nora

Abstract

MicroRNAs (miRNAs) are small non-coding endogenous RNAs, which are attracting a growing interest as therapeutic molecules due to their central role in major diseases. However, the transformation of these biomolecules into drugs is limited due to their unstability in the bloodstream, caused by nucleases abundantly present in the blood, and poor capacity to enter cells. The conjugation of miRNAs to nanoparticles (NPs) could be an effective strategy for their clinical delivery. Herein, the engineering of non-liposomal lipid nanovesicles, named quatsomes (QS), for the delivery of miRNAs and other small RNAs into the cytosol of tumor cells, triggering a tumor-suppressive response is reported. The engineered pH-sensitive nanovesicles have controlled structure (unilamellar), size (<150 nm) and composition. These nanovesicles are colloidal stable (>24 weeks), and are prepared by a green, GMP compliant, and scalable one-step procedure, which are all unavoidable requirements for the arrival to the clinical practice of NP based miRNA therapeutics. Furthermore, QS protect miRNAs from RNAses and when injected intravenously, deliver them into liver, lung, and neuroblastoma xenografts tumors. These stable nanovesicles with tunable pH sensitiveness constitute an attractive platform for the efficient delivery of miRNAs and other small RNAs with therapeutic activity and their exploitation in the clinics.

Published
2022

2. Engineering pH-Sensitive Stable Nanovesicles for Delivery of MicroRNA Therapeutics

Author

Dganit Danino, Alba Córdoba, Santi Sala, Ariadna Boloix, Lorenzo Albertazzi, Inbal Abutbul-Ionita, Mariana Köber, Marc Masanas, Soledad Gallego, Josep Sánchez de Toledo, Javier Repetto, Nora Ventosa, Nathaly Segovia, Natalia Feiner-Gracia, Miguel F. Segura, Aroa Soriano, Rosa Pascarella, Josep Merlo-Mas, Josep Roma, Jaume Veciana, Guillem Vargas-Nadal, Laia Foradada, Institut Català de la Salut, [Boloix A] Molecular Nanoscience and Organic Materials (Nanomol) Institut de Ciència de Materials de Barcelona ICMAB-CSIC Campus UAB. Laboratori de Recerca Translacional en Càncer en la Infància i l’Adolescència, Vall d’Hebron Institut de Recerca (VHIR), Barcelona, Spain. Universitat Autònoma de Barcelona, Bellaterra, Spain. CIBER de Bioingeniería Biomateriales y Nanomedicina (CIBER-BBN) Madrid, Spain. [Feiner-Gracia N, Pascarella R] Nanoscopy for Nanomedicine Group Institute for Bioengineering of Catalonia (IBEC) The Barcelona Institute of Science and Technology (BIST) Barcelona, Spain. Department of Biomedical Engineering Institute for Complex Molecular Systems (ICMS) Eindhoven University of Technology Eindhoven, The Netherlands. [Köber M] Molecular Nanoscience and Organic Materials (Nanomol) Institut de Ciència de Materials de Barcelona ICMAB-CSIC Campus UAB. CIBER de Bioingeniería Biomateriales y Nanomedicina (CIBER-BBN) Madrid, Spain. [Repetto J] Molecular Nanoscience and Organic Materials (Nanomol) Institut de Ciència de Materials de Barcelona ICMAB-CSIC Campus UAB. [Soriano A, Masanas M, Roma J, Sánchez de Toledo J, Gallego S] Laboratori de Recerca Translacional en Càncer en la Infància i l’Adolescència, Vall d’Hebron Institut de Recerca (VHIR), Barcelona, Spain. Universitat Autònoma de Barcelona, Bellaterra, Spain. [Foradada L] Peptomyc S.L., Edifici Cellex Barcelona, Spain. [Segura MF] Molecular Nanoscience and Organic Materials (Nanomol) Institut de Ciència de Materials de Barcelona ICMAB-CSIC Campus UAB. Laboratori de Recerca Translacional en Càncer en la Infància i l’Adolescència, Vall d’Hebron Institut de Recerca (VHIR), Barcelona, Spain. Universitat Autònoma de Barcelona, Bellaterra, Spain, Vall d'Hebron Barcelona Hospital Campus, Ministerio de Educación, Cultura y Deporte (España), Ministerio de Economía y Competitividad (España), Ministerio de Ciencia, Innovación y Universidades (España), Instituto de Salud Carlos III, Centro de Investigación Biomédica en Red Bioingeniería, Biomateriales y Nanomedicina (España), Matem lo Bitxo, Asociación Española Contra el Cáncer, Generalitat de Catalunya, Boloix, Ariadna, Köber, Mariana, Soriano, Aroa, Masanas, Marc, Segovia, Nathaly, Vargas Nadal, Guillem, Merlo Mas, Josep, Danino, Dganit, Foradada, Laia, Roma, Josep, Toledo, Josep Sánchez de, Gallego, Soledad, Veciana, Jaume, Albertazzi, Lorenzo, Segura, Miguel F., Ventosa, Nora, ICMS Core, Nanoscopy for Nanomedicine, Molecular Biosensing for Med. Diagnostics, Boloix, Ariadna [0000-0002-1648-5589], Köber, Mariana [0000-0001-9962-7900], Soriano, Aroa [0000-0001-9659-1471], Masanas, Marc [0000-0002-2249-8554], Segovia, Nathaly [0000-0001-8814-6095], Vargas Nadal, Guillem [0000-0003-4383-1325], Merlo Mas, Josep [0000-0002-3698-6655], Danino, Dganit [0000-0002-9782-4940], Foradada, Laia [0000-0002-0589-4360], Roma, Josep [0000-0001-7692-6123], Toledo, Josep Sánchez de [0000-0002-1034-1920], Gallego, Soledad [0000-0002-4712-9624], Veciana, Jaume [0000-0003-1023-9923], Albertazzi, Lorenzo [0000-0002-6837-0812], Segura, Miguel F. [0000-0003-0916-3618], and Ventosa, Nora [0000-0002-8008-4974]

Subjects
Micro RNAs, Cancer therapy, nanovesicles, siRNAs delivery, Otros calificadores::Otros calificadores::/farmacoterapia [Otros calificadores], Endogeny, Tumor cells, SDG 3 – Goede gezondheid en welzijn, Other subheadings::Other subheadings::/drug therapy [Other subheadings], Nanovesicles, Biomaterials, neoplasias [ENFERMEDADES], neuroblastoma, SDG 3 - Good Health and Well-being, Neuroblastoma, Neoplasms, microRNA, quatsomes, medicine, Humans, General Materials Science, Tumors, miRNAs delivery, MicroARN, Nanopartícules, nanocarriers, Chemistry, Càncer - Tractament, General Chemistry, Hydrogen-Ion Concentration, medicine.disease, Pediatric cancer, Cell biology, pediatric cancer, Clinical Practice, Neoplasms [DISEASES], Cytosol, MicroRNAs, cancer therapy, Nanoparticles, Nanocarriers, Biotechnology
Abstract

MicroRNAs (miRNAs) are small non-coding endogenous RNAs, which are attracting a growing interest as therapeutic molecules due to their central role in major diseases. However, the transformation of these biomolecules into drugs is limited due to their unstability in the bloodstream, caused by nucleases abundantly present in the blood, and poor capacity to enter cells. The conjugation of miRNAs to nanoparticles (NPs) could be an effective strategy for their clinical delivery. Herein, the engineering of non-liposomal lipid nanovesicles, named quatsomes (QS), for the delivery of miRNAs and other small RNAs into the cytosol of tumor cells, triggering a tumor-suppressive response is reported. The engineered pH-sensitive nanovesicles have controlled structure (unilamellar), size (24 weeks), and are prepared by a green, GMP compliant, and scalable one-step procedure, which are all unavoidable requirements for the arrival to the clinical practice of NP based miRNA therapeutics. Furthermore, QS protect miRNAs from RNAses and when injected intravenously, deliver them into liver, lung, and neuroblastoma xenografts tumors. These stable nanovesicles with tunable pH sensitiveness constitute an attractive platform for the efficient delivery of miRNAs and other small RNAs with therapeutic activity and their exploitation in the clinics., The funding was received by Ministerio de Educación, Cultura y Deporte (Grant no. FPU16/01099), Ministerio de Economía, Industria y Competividad (Grants MAT2016-80820-R, MAT2016-80826-R and SAF2016-75241-R), the Ministry of Science and Innovation (MINECO) of Spain through grant PID2019-105622RB-I00, from Instituto de Salud Carlos III (Grant no. CP16/00006, PI17/00564, PI20/00530, DTS20/00018) (Co-funded by European Regional Development Fund/European Social Fund) “Investing in your future”), from the EuroNanoMed II platform through the NanoVax project, from CIBER-BBN through grant TAG-SMARTLY, Joan Petit Foundation, Asociación Matem Lo Bitxo and Asociación Española Contra el Cáncer (Grant no. LABAE18009SEGU), as well as, Generalitat de Catalunya through the Centres de Recerca de Catalunya (CERCA) programme and grant no. 2017-SGR-918, and from Agency for Management of University and Research Grants (AGAUR) (Grant no 2018LLAV0064 and SIFECAT IU68-010017). Furthermore, ICMAB-CSIC acknowledges support from the MINECO through the Severo Ochoa Programme for Centres of Excellence in R&D (SEV-2015-0496 and CEX2019-000917-S). Quatsome production and their physicochemical characterization was performed by the ICTS “NANBIOSIS,” more specifically in the Biomaterial Processing and Nanostructuring Unit (U6), Unit of the CIBER in Bioengineering, Biomaterials & Nanomedicne (CIBER-BBN) located at the Institute of Materials Science of Barcelona (ICMAB-CSIC). The authors thank the UAB Microscopy service for their help in recording cryo-TEM images. The authors also thank Mr. Adolfo de Hoyos-Limon for pKa measurements, Ms. Patricia Pérez for her help in in vitro experiments, the members of Laboratory Animal Service Unit of Vall d'Hebron Research Institute for their help in the in vivo experiment. The authors thank Editage (www.editage.com) and Ms. Christine O'Hara for English language correction. The authors acknowledge Biorender.com for allowing the adaptation of their templates. Retrieved from https://app.biorender.com/biorender-templates., With funding from the Spanish government through the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000917-S).

Published
2021

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