Graphene nanoribbons elicit cell specific uptake and delivery via activation of epidermal growth factor receptor enhanced by human papillomavirus E5 protein

Abstract

Ligands such as peptides, antibodies or other epitopes bind and activate specific cell receptors, and are employed for targeted cellular delivery of pharmaceuticals such as drugs, genes and imaging agents. Herein, we show that oxidized graphene nanoribbons, non-covalently functionalized with PEG-DSPE (1, 2-distearoyl-sn-glycero-3-phosphoethanolamine-N[amino(polyethyleneglycol)]) (O-GNR-PEG-DSPE) activate epidermal growth factor receptors (EGFRs). This activation generates predominantly dynamin-dependent macropinocytosis-like response, and results in significant O-GNR-PEG-DSPE uptake into cells with high EGFR expression. Cells with an integrated human papillomavirus (HPV) genome also show increased uptake due to the modulation of the activated EFGR by the viral protein E5. We demonstrate that this cell specific uptake of O-GNR-PEG-DSPE can be exploited to achieve significantly enhanced drug efficacies even in drug resistant cells. These results have implications towards the development of active targeting and delivery agents without ligand functionalization for use in the diagnosis and treatment of pathologies that overexpress EGFR or mediated by HPV.

Graphene Enhances Cellular Proliferation through Activating the Epidermal Growth Factor Receptor

“…Similar results in the study by Mullick Chowdhury et al showed that A549 cells had no uptake of graphene nanoribbons. 32 Jaworski et al also pointed out that graphene platelets did not enter into the cells. 33 Nevertheless, as seen from Figure 4 and panels B−D of Figure 5, plenty of GS deposited closely to or attached on the cell membrane (indicated by the yellow arrow), which could pose direct stimulations to the cells through the physical contact.…”

Interactions of 1D- and 2D-Layered Inorganic Nanoparticles with Fibroblasts and Human Mesenchymal Stem Cells

“…Outside the cell, MSNPs and WSNTs were observed on the cell membranes. However, cytoplasmic protrusions observed during micropinocytosis were seen only around the WSNT aggregates [37]. These observations suggest that WSNTs and MSNPs can enter the cells via different uptake mechanisms without affecting viability or differentiation potential of MSCs at concentrations up to 50 µg/ml.…”

Delivery of long chain C16 and C24 ceramide in HeLa cells using oxidized graphene nanoribbons

“…Because C 16 and C 24 ceramides are native components of cellular membranes, we wanted to determine if they had an effect on the entry of O‐GNRs into cells. We have previously established that O‐GNRs are uptaken into HeLa cells via a macropinocytotic‐like, dynamin‐dependent mechanism that relies on activation of epidermal growth factor receptor (EGFR) (Mullick Chowdhury et al, ). This was determined using four different inhibitors of specific uptake pathways at non‐lethal concentrations: the dynamin inhibitor dynasore, the EGFR inhibitor gefitinib, the macropinocytosis inhibitor ethylisopropylamiloride (EIPA), and the caveolae‐mediated endocytosis inhibitor filipin.…”

“…We also wanted to determine if the loading of ceramide onto O‐GNRs would have a significant effect on their entry into cells. Previously we determined that O‐GNRs have a macropinocytotic‐like, dynamin‐dependent mechanism that relies on activation of EGFR (Mullick Chowdhury et al, ). Because ceramides are important components of cellular membranes, we wanted to determine if loading ceramides onto O‐GNRs would have an effect on their uptake.…”

“…It has also been shown that aliphatic hydrocarbons are capable of binding to these hydrophobic regions. The surfactant 1,2‐distearoyl‐ sn ‐glycero‐3‐phosphoethanolamine‐N‐[amino(polyethylene glycol)] (DSPE‐PEG) has been used to increase the dispersibility of graphene oxide nanoparticles in aqueous solution, based on the principle that the lipid moiety DSPE binds to the hydrophobic regions of graphene while the PEG moiety extends out into solution for increased solubility (Chowdhury et al, ; Mullick Chowdhury et al, ; Mullick Chowdhury, Manepalli, & Sitharaman, ). Thus, it is possible for an extremely hydrophobic molecule like long chain ceramides to bind to these hydrophobic regions.…”