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Carrier-Free CXCR4-Targeted Nanoplexes Designed for Polarizing Macrophages to Suppress Tumor Growth

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Abstract

Introduction

Treatment options for cancer metastases, the primary cause of cancer mortality, are limited. The chemokine receptor CXCR4 is an attractive therapeutic target in cancer because it mediates metastasis by inducing cancer cell and macrophage migration. Here we engineered carrier-free CXCR4-targeting RNA-protein nanoplexes that not only inhibited cellular migration but also polarized macrophages to the M1 phenotype.

Materials and Methods

A CXCR4-targeting single-chain variable fragment (scFv) antibody was fused to a 3030 Da RNA-binding protamine peptide (RSQSRSRYYRQRQRSRRRRRRS). Self-assembling nanoplexes were formed by mixing the CXCR4-scFv-protamine fusion protein (CXCR4-scFv-RBM) with miR-127-5p, a miRNA shown to mediate M1 macrophage polarization. RNA-protein nanoplexes were characterized with regard to their physicochemical properties and therapeutic efficacy.

Results

CXCR4-targeting RNA-protein nanoplexes simultaneously acted as a targeting ligand, a macrophage polarizing drug, and a miRNA delivery vehicle. Our carrier-free, RNA-protein nanoplexes specifically bound to CXCR4-positive macrophages and breast cancer cells, showed high drug loading (~ 90% w/w), and are non-toxic. Further, these RNA-protein nanoplexes significantly inhibited cancer and immune cell migration (75 to 99%), robustly polarized macrophages to the tumor-suppressive M1 phenotype, and inhibited tumor growth in a mouse model of triple-negative breast cancer

Conclusions

We engineered a novel class of non-toxic RNA-protein nanoplexes that modulate the tumor stroma. These nanoplexes are promising candidates for add-ons to clinically approved chemotherapeutics.

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Abbreviations

scFv:

Single-chain variable fragment

RBM:

RNA-binding motif

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Acknowledgments

We acknowledge support from the NIH through Awards EB023262 and EB021454. We thank Dr. Mark Sullivan for providing us with the phage display library. We thank Dr. Brian Kay and Dr. Renhua Huang for training in techniques related to phage display and screening. We thank Dr. Gauri Rao for helpful discussions.

Conflict of interest

All authors, including M.B. Deci, M. Liu, J. Gonya, C.J. Lee, Tingyi Li, S.W. Ferguson, E.E. Bonacquisti, J. Wang, and J. Nguyen, declare that they have no conflicts of interest.

Ethical Approval

All institutional and national guidelines for the care and use of laboratory animals were followed and approved by the appropriate institutional committees. No human studies were carried out by the authors for this article

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Author notes

  1. Michael B. Deci and Maixian Liu contributed equally to this work.

Authors and Affiliations

  1. Department of Pharmaceutical Sciences, School of Pharmacy, University at Buffalo, The State University of New York, Buffalo, NY, 14214, USA

    Michael B. Deci, Maixian Liu, Jacqueline Gonya, Christine J. Lee, Scott W. Ferguson, Jinli Wang & Juliane Nguyen

  2. Division of Pharmacoengineering and Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA

    Tingyi Li, Emily E. Bonacquisti & Juliane Nguyen

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  1. Michael B. Deci
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Correspondence to Juliane Nguyen.

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Juliane Nguyen is an Associate Professor in the Division of Pharmacoengineering and Molecular Therapeutics at the University of North Carolina at Chapel Hill. She received her PharmD and PhD in Pharmaceutical Sciences from the Philipps-University of Marburg (2009), where she trained with Professor Thomas Kissel. From 2009 to 2013, she was a Deutsche Forschungsgemeinschaft postdoctoral fellow in Professor Francis Szoka’s laboratory at the University of California, San Francisco. Dr. Nguyen started her laboratory at the University at Buffalo in July 2013 and was promoted to Associate Professor in January 2019. Her laboratory develops novel protein-, RNA-, and lipid-based biochemical and delivery platforms for treating myocardial infarction and cancer. Dr. Nguyen is the recipient of numerous awards, including the Biomedical Breakthrough Award (2011), the University at Buffalo Exceptional Scholar Young Investigator Award (2017), the Pioneering Pharmaceutical Sciences by Emerging Investigator Award (2018), the NSF CAREER Award (2018), and the NYSTAR faculty award (2019).

This article is part of the CMBE 2019 Young Innovators special issue.

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Deci, M.B., Liu, M., Gonya, J. et al. Carrier-Free CXCR4-Targeted Nanoplexes Designed for Polarizing Macrophages to Suppress Tumor Growth. Cel. Mol. Bioeng. 12, 375–388 (2019). https://doi.org/10.1007/s12195-019-00589-w

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