Nano-encapsulation of a novel anti-Ran-GTPase peptide for blockade of regulator of chromosome condensation 1 (RCC1) function in MDA-MB-231 breast cancer cells

Yusuf Haggag, Kyle Matchett, El-Habib Dakir, Paul Buchanan, Mohammed Osman, Sanaa Elgizawy, Mohamed El Tanani, Ahmed Faheem, Paul McCarron

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Ran is a small ras-related GTPase and is highly expressed in aggressive breast carcinoma. Overexpression induces malignant transformation and drives metastatic growth. We have designed a novel series of anti-Ran-GTPase peptides, which prevents Ran hydrolysis and activation, and although they display effectiveness in silico, peptide activity is suboptimal in vitro due to reduced bioavailability and poor delivery. To overcome this drawback, we delivered an anti-Ran-GTPase peptide using encapsulation in PLGA-based nanoparticles (NP). Formulation variables within a double emulsion solvent evaporation technique were controlled to optimise physicochemical properties. NP were spherical and negatively charged with a mean diameter of 182–277 nm. Peptide integrity and stability were maintained after encapsulation and release kinetics followed a sustained profile. We were interested in the relationship between cellular uptake and poly(ethylene glycol) (PEG) in the NP matrix, with results showing enhanced in vitro uptake with increasing PEG content. Peptide-loaded, pegylated (10% PEG)-PLGA NP induced significant cytotoxic and apoptotic effects in MDA-MB-231 breast cancer cells, with no evidence of similar effects in cells pulsed with free peptide. Western blot analysis showed that encapsulated peptide interfered with the proposed signal transduction pathway of the Ran gene. Our novel blockade peptide prevented Ran activation by blockage of regulator of chromosome condensation 1 (RCC1) following peptide release directly in the cytoplasm once endocytosis of the peptide-loaded nanoparticle has occurred. RCC1 blockage was effective only when a nanoparticulate delivery approach was adopted.
LanguageEnglish
Pages40-53
JournalInternational Journal of Pharmaceutics
Volume521
Early online date2 Feb 2017
DOIs
Publication statusPublished - 15 Apr 2017

Fingerprint

Chromosomes, Human, Pair 1
GTP Phosphohydrolases
Breast Neoplasms
Peptides
Nanoparticles
Monomeric GTP-Binding Proteins
Ethylene Glycol
Endocytosis
Emulsions
Computer Simulation
Biological Availability
Signal Transduction
Cytoplasm
Hydrolysis
Western Blotting

Keywords

  • Anti-Ran-GTPase peptide
  • Double emulsion
  • PLGA
  • Nanoparticle
  • Breast cancer
  • Drug delivery

Cite this

Haggag, Yusuf ; Matchett, Kyle ; Dakir, El-Habib ; Buchanan, Paul ; Osman, Mohammed ; Elgizawy, Sanaa ; El Tanani, Mohamed ; Faheem, Ahmed ; McCarron, Paul. / Nano-encapsulation of a novel anti-Ran-GTPase peptide for blockade of regulator of chromosome condensation 1 (RCC1) function in MDA-MB-231 breast cancer cells. 2017 ; Vol. 521. pp. 40-53.
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Nano-encapsulation of a novel anti-Ran-GTPase peptide for blockade of regulator of chromosome condensation 1 (RCC1) function in MDA-MB-231 breast cancer cells. / Haggag, Yusuf; Matchett, Kyle; Dakir, El-Habib; Buchanan, Paul; Osman, Mohammed; Elgizawy, Sanaa; El Tanani, Mohamed; Faheem, Ahmed; McCarron, Paul.

Vol. 521, 15.04.2017, p. 40-53.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Nano-encapsulation of a novel anti-Ran-GTPase peptide for blockade of regulator of chromosome condensation 1 (RCC1) function in MDA-MB-231 breast cancer cells

AU - Haggag, Yusuf

AU - Matchett, Kyle

AU - Dakir, El-Habib

AU - Buchanan, Paul

AU - Osman, Mohammed

AU - Elgizawy, Sanaa

AU - El Tanani, Mohamed

AU - Faheem, Ahmed

AU - McCarron, Paul

PY - 2017/4/15

Y1 - 2017/4/15

N2 - Ran is a small ras-related GTPase and is highly expressed in aggressive breast carcinoma. Overexpression induces malignant transformation and drives metastatic growth. We have designed a novel series of anti-Ran-GTPase peptides, which prevents Ran hydrolysis and activation, and although they display effectiveness in silico, peptide activity is suboptimal in vitro due to reduced bioavailability and poor delivery. To overcome this drawback, we delivered an anti-Ran-GTPase peptide using encapsulation in PLGA-based nanoparticles (NP). Formulation variables within a double emulsion solvent evaporation technique were controlled to optimise physicochemical properties. NP were spherical and negatively charged with a mean diameter of 182–277 nm. Peptide integrity and stability were maintained after encapsulation and release kinetics followed a sustained profile. We were interested in the relationship between cellular uptake and poly(ethylene glycol) (PEG) in the NP matrix, with results showing enhanced in vitro uptake with increasing PEG content. Peptide-loaded, pegylated (10% PEG)-PLGA NP induced significant cytotoxic and apoptotic effects in MDA-MB-231 breast cancer cells, with no evidence of similar effects in cells pulsed with free peptide. Western blot analysis showed that encapsulated peptide interfered with the proposed signal transduction pathway of the Ran gene. Our novel blockade peptide prevented Ran activation by blockage of regulator of chromosome condensation 1 (RCC1) following peptide release directly in the cytoplasm once endocytosis of the peptide-loaded nanoparticle has occurred. RCC1 blockage was effective only when a nanoparticulate delivery approach was adopted.

AB - Ran is a small ras-related GTPase and is highly expressed in aggressive breast carcinoma. Overexpression induces malignant transformation and drives metastatic growth. We have designed a novel series of anti-Ran-GTPase peptides, which prevents Ran hydrolysis and activation, and although they display effectiveness in silico, peptide activity is suboptimal in vitro due to reduced bioavailability and poor delivery. To overcome this drawback, we delivered an anti-Ran-GTPase peptide using encapsulation in PLGA-based nanoparticles (NP). Formulation variables within a double emulsion solvent evaporation technique were controlled to optimise physicochemical properties. NP were spherical and negatively charged with a mean diameter of 182–277 nm. Peptide integrity and stability were maintained after encapsulation and release kinetics followed a sustained profile. We were interested in the relationship between cellular uptake and poly(ethylene glycol) (PEG) in the NP matrix, with results showing enhanced in vitro uptake with increasing PEG content. Peptide-loaded, pegylated (10% PEG)-PLGA NP induced significant cytotoxic and apoptotic effects in MDA-MB-231 breast cancer cells, with no evidence of similar effects in cells pulsed with free peptide. Western blot analysis showed that encapsulated peptide interfered with the proposed signal transduction pathway of the Ran gene. Our novel blockade peptide prevented Ran activation by blockage of regulator of chromosome condensation 1 (RCC1) following peptide release directly in the cytoplasm once endocytosis of the peptide-loaded nanoparticle has occurred. RCC1 blockage was effective only when a nanoparticulate delivery approach was adopted.

KW - Anti-Ran-GTPase peptide

KW - Double emulsion

KW - PLGA

KW - Nanoparticle

KW - Breast cancer

KW - Drug delivery

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DO - 10.1016/j.ijpharm.2017.02.006

M3 - Article

VL - 521

SP - 40

EP - 53

ER -