Detection and Imaging of Aggressive Cancer Cells Using an Epidermal Growth Factor Receptor (EGFR)-Targeted Filamentous Plant Virus-Based Nanoparticle

Abstract

Molecular imaging approaches and targeted drug delivery hold promise for earlier detection of diseases and treatment with higher efficacy while reducing side effects, therefore increasing survival rates and quality of life. Virus-based nanoparticles are a promising platform because their scaffold can be manipulated both genetically and chemically to simultaneously display targeting ligands while carrying payloads for diagnosis or therapeutic intervention. Here, we displayed a 12-amino-acid peptide ligand, GE11 (YHWYGYTPQNVI), on nanoscale filaments formed by the plant virus potato virus X (PVX). Bioconjugation was used to produce fluorescently labeled PVX-GE11 filaments targeted toward the epidermal growth factor receptor (EGFR). Cell detection and imaging was demonstrated using human skin epidermoid carcinoma, colorectal adenocarcinoma, and triple negative breast cancer cell lines (A-431, HT-29, MDA-MB-231), all of which upregulate EGFR to various degrees. Nonspecific uptake in ductal breast carcinoma (BT-474) cells was not observed. Furthermore, co-culture experiments with EGFR+ cancer cells and macrophages indicate successful targeting and partitioning toward the cancer cells. This study lays a foundation for the development of EGFR-targeted filaments delivering contrast agents for imaging and diagnosis, and/or toxic payloads for targeted drug delivery.

Design of virus-based nanomaterials for medicine, biotechnology, and energy

“…Some examples of targets that have been used for specific uptake of virus-based nanoparticles include epidermal growth factor receptor (EGFR) 161, 162 and folate receptor (FR). 163, 164 In such a manner, overexpressed receptors or environmental cues can be tracked for diagnostic or drug delivery purposes, which will be discussed in the following sections.…”

“…Strategies range from display of EGF on Qβ through genetic engineering, 162 using phage antibody libraries to select for fd phages with single-chain antibody variable fragments (scFvs) specific for EGFR as well as its related receptor human epidermal growth factor receptor 2 (HER2), 101 conjugation of EGFR antibodies on MS2, 188 and also chemical attachment of GE11 peptide on PVX. 161 These studies all evaluated cell binding in vitro and there are some promising results indicating partitioning of targeted particles to tumor cells compared to macrophages in co-cultures, 161 and it would be of interest to see their development in mouse models.…”

Potato virus X, a filamentous plant viral nanoparticle for doxorubicin delivery in cancer therapy

“…22 PVX has been shown to target to tumors by passive accumulation via the enhanced permeability and retention (EPR) effect 23 or active targeting by means of presentation of receptor-specific ligands. 24 Analogous to other elongated materials, PVX exhibits enhanced tumor homing and tissue penetration compared to the spherical cowpea mosaic virus (CPMV) in a variety of cancer models including human tumor xenografts of fibrosarcoma, colon cancer, and breast cancer. 23,25 We also demonstrated that the in vivo fate of PVX could be further enhanced by conjugation of stealth polymers such as polyethylene glycol (PEG) via the solvent-exposed lysine side chains.…”

The Protein Corona of Plant Virus Nanoparticles Influences their Dispersion Properties, Cellular Interactions, and In Vivo Fates

“…Some design principles for VNPs have been established: shape engineering allows the avoidance of non‐specific sequestration in cells of the mononuclear phagocyte system (MPS), polymer coatings reduce the immunogenicity of VNPs, and the addition of targeting ligands can increase cell specificity . Nevertheless, the fundamental interactions between native or functionalized VNPs and plasma proteins, and the impact of such interactions on biodistribution, clearance, and molecular target recognition, are poorly understood.…”