Intracellular delivery of recombinant arginine deiminase (rADI) by heparin-binding hemagglutinin adhesion peptide restores sensitivity in rADI-resistant cancer cells.

Abstract

Recombinant arginine deiminase (rADI) has been used in clinical trials for arginine-auxotrophic cancers. However, the emergence of rADI resistance, due to the overexpression of argininosuccinate synthetase (AS), has introduced an obstacle in its clinical application. Here, we have proposed a strategy for the intracellular delivery of rADI, which depletes both extracellular and intracellular arginine, to restore the sensitivity of rADI-resistant cancer cells. In this study, the C terminus of heparin-binding hemagglutinin adhesion protein from Mycobacterium tuberculosis (HBHAc), which contains 23 amino acids, was used to deliver rADI into rADI-resistant human breast adenocarcinoma cells (MCF-7). Chemical conjugates (l- and d-HBHAc-SPDP-rADI) and a recombinant fusion protein (rHBHAc-ADI) were produced. l- and d-HBHAc-SPDP-rADI showed a significantly higher cellular uptake of rADI by MCF-7 cells compared to that of rADI alone. Cell viability was significantly decreased in a dose-dependent manner in response to l- and d-HBHAc-SPDP-rADI treatments. In addition, the ratio of intracellular concentration of citrulline to arginine in cells treated with l- and d-HBHAc-SPDP-rADI was significantly increased by 1.4- and 1.7-fold, respectively, compared with that obtained in cells treated with rADI alone (p < 0.001). Similar results were obtained with the recombinant fusion protein rHBHAc-ADI. Our study demonstrates that the increased cellular uptake of rADI by HBHAc modification can restore the sensitivity of rADI treatment in MCF-7 cells. rHBHAc-ADI may represent a novel class of antitumor enzyme with an intracellular mechanism that is independent of AS expression.

• Authors: Fe-Lin Lin Wu, Tzyy-Harn Yeh, Ying-Luen Chen, Yu-Chin Chiu, Ju-Chen Cheng, Ming-Feng Wei, Li-Jiuan Shen
• Journal: Molecular pharmaceutics (Mol Pharm) Vol. 11 Issue 8 Pg. 2777-86 (Aug 04 2014) ISSN: 1543-8392 [Electronic] United States
• PMID: 24950134 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Amino Acids
• Lectins
• Peptides
• Recombinant Fusion Proteins
• heparin-binding hemagglutinin
• Citrulline
• Hydrolases
• arginine deiminase
• Argininosuccinate Synthase
• Fluorescein

Topics

• Amino Acids (chemistry)
• Argininosuccinate Synthase (metabolism)
• Cell Line, Tumor
• Cell Survival
• Citrulline (chemistry)
• Cytoplasm (metabolism)
• Dose-Response Relationship, Drug
• Drug Delivery Systems
• Drug Resistance, Neoplasm (drug effects)
• Endocytosis
• Fluorescein (chemistry)
• Humans
• Hydrolases (administration & dosage)
• Lectins (chemistry)
• MCF-7 Cells
• Mycobacterium tuberculosis
• Neoplasms (drug therapy)
• Peptides (chemistry)
• Recombinant Fusion Proteins (administration & dosage)