Norepinephrine transporter-derived homing peptides enable rapid endocytosis of drug delivery nanovehicles into neuroblastoma cells

Abstract

Background: Currently, the diagnosis and treatment of neuroblastomas-the most frequent solid tumors in children-exploit the norepinephrine transporter (hNET) via radiolabeled norepinephrine analogs. We aim to develop a nanomedicine-based strategy towards precision therapy by targeting hNET cell-surface protein with hNET-derived homing peptides. Results: The peptides (seq. GASNGINAYL and SLWERLAYGI) were shown to bind high-resolution homology models of hNET in silico. In particular, one unique binding site has marked the sequence and structural similarities of both peptides, while most of the contribution to the interaction was attributed to the electrostatic energy of Asn and Arg (< − 228 kJ/mol). The peptides were comprehensively characterized by computational and spectroscopic methods showing ~ 21% β-sheets/aggregation for GASNGINAYL and ~ 27% α-helix for SLWERLAYGI. After decorating 12-nm ferritin-based nanovehicles with cysteinated peptides, both peptides exhibited high potential for use in actively targeted neuroblastoma nanotherapy with exceptional in vitro biocompatibility and stability, showing minor yet distinct influences of the peptides on the global expression profiles. Upon binding to hNET with fast binding kinetics, GASNGINAYLC peptides enabled rapid endocytosis of ferritins into neuroblastoma cells, leading to apoptosis due to increased selective cytotoxicity of transported payload ellipticine. Peptide-coated nanovehicles significantly showed higher levels of early apoptosis after 6 h than non-coated nanovehicles (11% and 7.3%, respectively). Furthermore, targeting with the GASNGINAYLC peptide led to significantly higher degree of late apoptosis compared to the SLW-ERLAYGIC peptide (9.3% and 4.4%, respectively). These findings were supported by increased formation of reactive oxygen species, down-regulation of survivin and Bcl-2 and up-regulated p53. Conclusion: This novel homing nanovehicle employing GASNGINAYLC peptide was shown to induce rapid endocytosis of ellipticine-loaded ferritins into neuroblastoma cells in selective fashion and with successful payload. Future homing peptide development via lead optimization and functional analysis can pave the way towards efficient peptide-based active delivery of nanomedicines to neuroblastoma cells.

Prediction to the prognosis of children with neuroblastoma by nomogram based on the first-diagnosed inflammatory markers

“…A previous ferritin study of a large sample of neuroblastoma patients found that ferritin had a strong impact on the prognosis of high-risk NB patients receiving modern treatment after 2009, and ferritin show promise for (1) identifying ultra-high-risk; (2) re ning risk strati cation; and (3) clinical utility in low-/middle-income countries [16]. In addition, ferritin plays an important role in peptide-based active delivery of nanomedicines to neuroblastoma cells [17] . This is consistent with the ndings of our study.…”

Metallothionein-3 is a multifunctional driver that modulates the development of sorafenib-resistant phenotype in hepatocellular carcinoma cells

“…Label-free analysis of the effect of hMT3 up-regulation on proliferation of Huh7 cells was performed using real-time electrical impedance measurement on the xCELLigence RTCA DP instrument (Roche Diagnostics GmbH, Basel, Switzerland) according to a previous study [ 33 ].…”

Accelerating therapeutic protein design with computational approaches toward the clinical stage

“…Protein-based artificial nanosystem is another novel protein-based therapeutic with the requirement of high affinity. Yazan Haddad and co-workers developed an ellipticine-loaded ferritin with surface modification of the norepinephrine transporter (hNET) targeting peptides for tumor-targeting ability [188] . With computational methods, homology modeling, molecular docking, and molecular dynamics in this research, they found that these peptides showed an intriguing binding affinity with hNET.…”