Cyclic Peptide-Gadolinium Nanocomplexes as siRNA Delivery Tools

Authors

Amir Nasrolahi Shirazi, Marshall B. Ketchum UniversityFollow
Muhammad Imran Sajid, Chapman UniversityFollow
Dindyal Mandal, Chapman UniversityFollow
David Stickley, Marshall B. Ketchum University
Stephanie Nagasawa, Marshall B. Ketchum University
Joshua Long, Marshall B. Ketchum University
Sandeep Lohan, Chapman UniversityFollow
Keykavous Parang, Chapman UniversityFollow
Rakesh Kumar Tiwari, Chapman UniversityFollow

Document Type

Article

Publication Date

10-20-2021

Abstract

We have recently reported that a cyclic peptide containing five tryptophan, five arginine, and one cysteine amino acids [(WR)₅C], was able to produce peptide-capped gadolinium nanoparticles, [(WR)₅C]-GdNPs, in the range of 240 to 260 nm upon mixing with an aqueous solution of GdCl₃. Herein, we report [(WR)₅C]-GdNPs as an efficient siRNA delivery system. The peptide-based gadolinium nanoparticles (50 µM) did not exhibit significant cytotoxicity (~93% cell viability at 50 µM) in human leukemia T lymphoblast cells (CCRF-CEM) and triple-negative breast cancer cells (MDA-MB-231) after 48 h. Fluorescence-activated cell sorting (FACS) analysis indicated that the cellular uptakes of Alexa-488-labeled siRNA were found to be enhanced by more than 10 folds in the presence of [(WR)₅C]-GdNPs compared with siRNA alone in CCRF-CEM and MDA-MB-231 cells after 6 h of incubation at 37 °C. The gene silencing efficacy of the nanoparticles was determined via the western blot technique using an over-expressed gene, STAT-3 protein, in MDA-MB-231 cells. The results showed ~62% reduction of STAT-3 was observed in MDA-MB-231 with [(WR)₅C]-GdNPs at N/P 40. The integrity of the cellular membrane of CCRF-CEM cells was found to be intact when incubated with [(WR)₅C]-Gd nanoparticles (50 µM) for 2 h. Confocal microscopy reveals higher internalization of siRNA in MDA-MB-231 cells using [(WR)₅C]-GdNPs at N/P 40. These results provided insight about the use of the [(WR)₅C]-GdNPs complex as a potent intracellular siRNA transporter that could be a nontoxic choice to be used as a transfection agent for nucleic-acid-based therapeutics.

Comments

This article was originally published in Phamaceuticals, volume 14, issue 11, in 2021. https://doi.org/10.3390/ph14111064

Recommended Citation

Nasrolahi Shirazi A, Sajid MI, Mandal D, Stickley D, Nagasawa S, Long J, Lohan S, Parang K, Tiwari RK. Cyclic Peptide-Gadolinium Nanocomplexes as siRNA Delivery Tools. Pharmaceuticals. 2021; 14(11):1064. https://doi.org/10.3390/ph14111064

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The authors

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 License.