Humanized Lewis-Y specific antibody based delivery of STAT3 siRNA Journal Article
| Authors: | Ma, Y.; Kowolik, C. M.; Swiderski, P. M.; Kortylewski, M.; Yu, H.; Horne, D. A.; Jove, R.; Caballero, O. L.; Simpson, A. J. G.; Lee, F. T.; Pillay, V.; Scott, A. M. |
| Article Title: | Humanized Lewis-Y specific antibody based delivery of STAT3 siRNA |
| Abstract: | The clinical application of siRNA is limited largely by the lack of efficient, cell-specific delivery systems. Antibodies are attractive delivery vehicles for targeted therapy due to their high specificity. In this study we describe the use of a humanized monoclonal antibody (mAb), hu3S193, against Lewis-Y (Ley), as a delivery vehicle for STAT3 siRNA. This mAb is rapidly internalized into Ley-expressing cancer cells via antigen recognition, and when coupled to STAT3 siRNA, a potentially powerful molecularly targeted delivery agent is created. Selective silencing of STAT3 is associated with tumor suppression. Two hu3S193 based siRNA delivery systems using STAT3 siRNA as a prototype were developed and tested in Ley-positive cancer cells: (a) a covalent construct based on a reductive disulfide linker that is expected to undergo cleavage within cells and (b) a noncovalent construct based on (d-arginine)9 (9r) modified hu3S193. Ley-specific binding and internalization of both the covalent and noncovalent constructs were confirmed by flow cytometry and confocal microscopy. Both the covalent and the noncovalent system led to efficient STAT3 silencing in Ley-positive cancer cells (A431) but not in Ley-negative cancer cells (MDA-MB-435). The covalent construct, however, required co-treatment with reagents such as chloroquine or 9r that facilitate the escape of the siRNA from endosomes to achieve significant gene silencing. The 9r modified noncovalent construct induced ∼70% STAT3 knockdown at submicromolar siRNA concentrations when used at an optimal vehicle-to-siRNA ratio of 5:1. The STAT3 knockdown also led to ∼50% inhibition of cell proliferation of Ley-positive cells. Noncovalent linked STAT3 siRNA-hu3S193 has great promise for targeted knockdown of STAT3 in tumor cells. © 2011 American Chemical Society. |
| Keywords: | unclassified drug; human cell; flow cytometry; cell proliferation; gene targeting; stat3 protein; models, biological; confocal microscopy; small interfering rna; rna, small interfering; dose-response relationship, drug; cell line, tumor; structure-activity relationship; monoclonal antibody; drug delivery systems; antibodies, monoclonal; antigen; antigen recognition; stat3 transcription factor; molecular structure; gene silencing; internalization; arginine; lewis blood-group system; chloroquine; blood group lewis system; lewis y antigen; monoclonal antibody hu3s193 |
| Journal Title: | ACS Chemical Biology |
| Volume: | 6 |
| Issue: | 9 |
| ISSN: | 1554-8929 |
| Publisher: | American Chemical Society |
| Date Published: | 2011-09-16 |
| Start Page: | 962 |
| End Page: | 970 |
| Language: | English |
| DOI: | 10.1021/cb200176v |
| PROVIDER: | scopus |
| PUBMED: | 21766840 |
| PMCID: | PMC3831028 |
| DOI/URL: | |
| Notes: | --- - "Export Date: 2 November 2011" - "Source: Scopus" |
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