Horseradish Peroxidase-Functionalized Gold Nanoconjugates for Breast Cancer Treatment Based on Enzyme Prodrug Therapy

Abstract

Gema Vivo-Llorca,1– 3 Ángela Morellá-Aucejo,1– 3 Alba García-Fernández,1– 3 Paula Díez,1– 4 Antoni Llopis-Lorente,1,5 Mar Orzáez,3,6 Ramón Martínez-Máñez1– 4,7 1Instituto Interuniversitario de Investigación de Reconocimiento Molecular y Desarrollo Tecnológico (IDM), Universitat Politècnica de València, Universitat de València, València, Spain; 2CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Madrid, Spain; 3Unidad Mixta UPV-CIPF de Investigación de Mecanismos de Enfermedades y Nanomedicina, Valencia, Universitat Politècnica de València, Centro de Investigación Príncipe Felipe, València, Spain; 4Unidad Mixta de Investigación en Nanomedicina y sensores, Universitat Politènica de València, Instituto de Investigación Sanitaria la Fe, Valènica, Spain; 5Department of Chemical Engineering and Chemistry, Institute for Complex Molecular Systems, Eindhoven University of Technology, Eindhoven, The Netherlands; 6Centro de Investigación Príncipe Felipe, Laboratorio de Péptidos y Proteínas, València, Spain; 7Departamento de Química, Universitat Politècnica de València, València, SpainCorrespondence: Alba García-Fernández; Ramón Martínez-Máñez Email [email protected]; [email protected]: Breast cancer has the highest mortality rate among cancers in women. Patients suffering from certain breast cancers, such as triple-negative breast cancer (TNBC), lack effective treatments. This represents a clinical concern due to the associated poor prognosis and high mortality. As an approach to succeed over conventional therapy limitations, we present herein the design and evaluation of a novel nanodevice based on enzyme-functionalized gold nanoparticles to efficiently perform enzyme prodrug therapy (EPT) in breast cancer cells.Results: In particular, the enzyme horseradish peroxidase (HRP) – which oxidizes the prodrug indole-3-acetic acid (IAA) to release toxic oxidative species – is incorporated on gold nanoconjugates (HRP-AuNCs), obtaining an efficient nanoplatform for EPT. The nanodevice is biocompatible and effectively internalized by breast cancer cell lines. Remarkably, co-treatment with HRP-AuNCs and IAA (HRP-AuNCs/IAA) reduces the viability of breast cancer cells below 5%. Interestingly, 3D tumor models (multicellular tumor spheroid-like cultures) co-treated with HRP-AuNCs/IAA exhibit a 74% reduction of cell viability, whereas the free formulated components (HRP, IAA) have no effect.Conclusion: Altogether, our results demonstrate that the designed HRP-AuNCs nanoformulation shows a remarkable therapeutic performance. These findings might help to bypass the clinical limitations of current tumor enzyme therapies and advance towards the use of nanoformulations for EPT in breast cancer.Keywords: gold nanoconjugates, horseradish peroxidase, indole-3-acetic acid, enzyme prodrug therapy, breast cancer

Keywords

• gold nanoconjugates
• horseradish peroxidase
• indole-3-acetic acid
• enzyme prodrug therapy
• breast cancer