A versatile theranostic nanodevice based on an orthogonal bioconjugation strategy for efficient targeted treatment and monitoring of triple negative breast cancer

doi:10.1016/j.nano.2019.102120

Nanomedicine: Nanotechnology, Biology and Medicine

Volume 24, February 2020, 102120

https://doi.org/10.1016/j.nano.2019.102120 Get rights and content

Abstract

A novel chemical-based orthogonal bioconjugation strategy to produce tri-functionalized nanoparticles (NPs) an chemotherapy drug, doxorubicin (DOX), a near-infrared cyanine dye (Cy7) and CRGDK homing peptide, a peptide specifically binds to neuropilin-1 (Nrp-1) overexpressed on triple negative breast cancer (TNBC) cells, has been validated. These theranostic NPs have been evaluated in vitro and in vivo using an orthotopic xenotransplant mouse model using TNBC cells. In vitro assays show that theranostic NPs improve the therapeutic index in comparison with free DOX. Remarkably, in vivo studies showed preferred location of theranostic NPs in the tumor area reducing the volume at the same level than free DOX while presenting lower side effects. This multifunctionalized theranostic nanodevice based on orthogonal conjugation strategies could be a good candidate for the treatment and monitoring of Nrp-1 overexpressing tumors. Moreover, this versatile nanodevice can be easily adapted to treat and monitor different cancer types by adapting the conjugation strategy.

Graphical Abstract

An effective, safe and versatile nanodevice for in vivo theranostic to target tumor vasculature has been successfully developed. Herein, we reported a theranostic nanodevice based on orthogonal conjugation strategies for the multifunctionalization of polymeric NPs that contain a controlled amount of each one of the bioactive cargoes. As proof of concept, NPs carrying doxorubicin (DOX), near-infrared cyanine dye (Cy7) and a homing peptide (CRGDK), which can actively recognize a receptor overexpressed in triple negative breast cancer have been successfully validated in vitro using MDA-MB-231 tumor cell line and in vivo using an orthotopic breast cancer xenotransplant mouse model.

Section snippets

Preparation of theranostic nanoparticles (HP-Cy7-DOX-NPs (14))

All solvents and chemicals were purchased from Sigma-Aldrich. Double PEGylated and bifunctionalized NPs (Fmoc-Dde-NPs, 6) were obtained by using protocols previously described¹⁶ (see S.I.). Next, DOX-NPs (9) were obtained by carboxy functionalization and hydrazine treatment followed by DOX conjugation (see S.I.) Then trifunctionalized NPs were generated by treatment with Fmoc-lysine-Dde(OH) (15 equiv), Fmoc deprotection and fluorophore conjugation (see S.I.). Finally, COOH-Cy7-DOX-NPs (13) were

Preparation and characterization of NPs (HP-Cy7-DOX-NPs (14)) as theranostic probes

A chemical strategy was developed to prepare multifunctionalized polymeric NPs. A monodisperse population of aminomethyl polystyrene cross-linked NPs were prepared by dispersion polymerization using a previously reported protocol.¹⁷ Size distribution was determined by DLS showing homogeneous size and low polydispersity (460 nm with a PDI of 0.042). Zeta potential value was 81.2 ± 0.1 mV. The concentration of NPs (4 × 10⁶ NP/μL ± 1.44 × 10⁻⁵) was calculated according to the method developed by

Discussion

In this study, we reported for the first time a successfully developed theranostic nanodevice based on orthogonal conjugation strategies for the multifunctionalization of polymeric NPs. Theranostic NPs (HP-Cy7-DOX-NPs (14)) containing controlled amount of each one of the components has been successfully prepared. A selective receptor-mediated release of DOX has been successfully achieved in TNBC cells overexpressing Nrp-1 by CRGDK peptide conjugation.

We used an orthogonal strategy to achieve

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Breast cancer is a collation of malignancies that manifest in the mammary glands at the early stages. Among breast cancer subtypes, triple-negative breast cancer (TNBC) shows the most aggressive behavior, with apparent stemness features. Owing to the lack of response to hormone therapy and specific targeted therapies, chemotherapy remains the first line of the TNBC treatment. However, the acquisition of resistance to chemotherapeutic agents increase therapy failure, and promotes cancer recurrence and distant metastasis. Invasive primary tumors are the birthplace of cancer burden, though metastasis is a key attribute of TNBC-associated morbidity and mortality. Targeting the chemoresistant metastases-initiating cells via specific therapeutic agents with affinity to the upregulated molecular targets is a promising step in the TNBC clinical management. Exploring the capacity of peptides as biocompatible entities with the specificity of action, low immunogenicity, and robust efficacy provides a principle for designing peptide-based drugs capable of increasing the efficacy of current chemotherapy agents for selective targeting of the drug-tolerant TNBC cells. Here, we first focus on the resistance mechanisms that TNBC cells acquire to evade the effect of chemotherapeutic agents. Next, the novel therapeutic approaches employing tumor-targeting peptides to exploit the mechanisms of drug resistance in chemorefractory TNBC are described.
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Citation Excerpt :
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: Acknowledgements: This work was supported by grants from the Ministry of Economy and Competitiveness (MINECO, National Programme for Research Aimed at the Challenges of Society, project reference BIO2016-80519-R&FEDER funds, grant number MAT2015-62644.C2.2.R), by Ministerio de Ciencia, Innovación y Universidades grant number RTI2018-101309-B-C2 (FEDER Funds), the Health Institute Carlos III (ISCIII) (Projects of technological development in health - ref DTS18/00121) and the Chair “Doctors Galera-Requena in cancer stem cell research”. This research was additionally supported by the University of Granada – PLAN OF RESEARCH AND TRANSFER 2016 –PSETC- PSE/16/003). The authors thank the Research Results Transfer Office (OTRI) of the University of Granada for their support for the technological development of this project. S.A.N-M. acknowledges the MINECO for providing a PhD fellowship (FPI) through the project MAT2015-62644.C2.2.R.

: Conflicts of interest: There are no conflicts to declare.

¹: Both authors contributed equally.

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Original ArticleA versatile theranostic nanodevice based on an orthogonal bioconjugation strategy for efficient targeted treatment and monitoring of triple negative breast cancer

Abstract

Graphical Abstract

Section snippets

Preparation of theranostic nanoparticles (HP-Cy7-DOX-NPs (14))

Preparation and characterization of NPs (HP-Cy7-DOX-NPs (14)) as theranostic probes

Discussion

Biomaterials

Bioconjug Chem

Anal Biochem

Anal Biochem

Biomaterials

Biomed Pharmacother

Curr Opin Chem Biol

Polymeric nanostructures for imaging and therapy

Chem Rev

Nanomaterials for theranostics: recent advances and future challenges

Chem Rev

Redox and pH dual responsive polymer based nanoparticles for in vivo drug delivery

Small

Enhancement of tumor homing by chemotherapy-loaded nanoparticles

Small

The use of solid supports to generate nucleic acid carriers

Acc Chem Res

Sulfhydryl reactive microspheres for the efficient delivery of thiolated bioactive cargoes

J Mater Chem

Polystyrene nanoparticles as a platform for studying nano interactions

Beilstein J Nanotechnol

Novel strategy for microsphere-mediated DNA transfection

Bioconjug Chem

Palladium-mediated intracellular chemistry

Nat Chem

Investigation of microsphere-mediated cellular delivery by chemical, microscopic and gene expression analysis

Mol Biosyst

Microsphere-based intracellular sensing of Caspase-3/7 in apoptotic living cells

Macromol Biosci

Tracking cell proliferation using a nanotechnology-based approach

Nanomedicine

associated molecules in the blood distinguish poor prognosis breast Cancer: a cross-sectional study

Sci Rep

Original Article
A versatile theranostic nanodevice based on an orthogonal bioconjugation strategy for efficient targeted treatment and monitoring of triple negative breast cancer