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In vitro and in vivo Evaluation of 111In-labeled E. coli Heat-Stable Enterotoxin Analogs for Specific Targeting of Human Breast Cancers

Breast Cancer Research and Treatment volume 98, pages 7–15 (2006)Cite this article

Summary

Research into the interaction between the E. coli heat-stable enterotoxin (STh) and the guanylin receptor guanylate cyclase C (GC-C) has generated >100 synthetic analogs of the peptide, several of which have been investigated as imaging or therapeutic agents for colorectal cancers. The evidence presented here suggests that in addition to STh binding to GC-C expressing cell lines derived from human colon, STh also specifically binds to an as yet unidentified receptor expressed in high densities on the surface of cell lines derived from human breast cancers. In vitro whole-cell crosslinking studies using 125I-labeled F19-STh(1–19) demonstrate that the putative STh binding protein migrates as an approximately 120–125 kDa species by SDS-PAGE, significantly smaller than the glycosylated GC-C molecule found in the T84 human colon cancer cell line. RT-PCR using total RNA isolated from breast and colon cancer cell lines indicates that GC-C transcripts are undetectable in human breast cancer cell lines and abundant in human colon cancer cell lines. In vitro competitive binding studies using STh analogs and the estrogen receptor positive (ER+) T-47D cell line demonstrated IC50 values between 2.6 and 8.5 nM. Similar studies on the estrogen receptor negative (ER−) cell line MDA-MB-231 showed IC50’s between 5.6 and 9.9 nM. Saturation binding analysis revealed receptor expression to fall between 40,000 and 120,000 sites per cell in these cell lines, receptor abundances equal to or greater than the abundance of GC-C in colorectal cancer cell lines. STh binding to these cells, although of similar affinity to STh binding to GC-C, is distinguishable from it on the basis of its ligand specificity. The characteristics of STh analogs as radiopharmaceutical agents were tested in an in vivo model utilizing T-47D human breast cancer cell xenografts in SCID mice. Clearance of STh analogs was rapid, primarily via renal excretion into the urine, with >85% ID excreted into the urine at 1 h p.i. Tumor uptake at 1 h p.i. in T-47D tumor cell xenografts was 0.67±0.23% ID/g, and was significantly decreased (p<0.05) upon co-administration of 4 mg/kg unlabeled STh. These results suggest that STh may find application for the imaging and treatment of breast cancer.

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Acknowledgements

This material is the result of work supported with resources and the use of facilities at the Harry S Truman Memorial Veterans’ Administration Hospital, Columbia, MO 65201, and the University of Missouri, Columbia School of Medicine Department of Radiology, Columbia, MO 65211. This work was supported by a grant from the National Cancer Institute (R01-CA95075), and by a National Cancer Institute Center grant (1 P50 CA103130-01).

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Affiliations

  1. Departments of Radiology, University of Missouri-Columbia, Columbia, MO, USA

    Michael F. Giblin, Hariprasad Gali, Nellie K. Owen & Wynn A. Volkert

  2. The Radiopharmaceutical Sciences Institute, University of Missouri-Columbia, Columbia, MO, USA

    Michael F. Giblin, Timothy J. Hoffman, Wynn A. Volkert & Leonard R. Forte

  3. Research Service, Harry S. Truman Memorial Veterans’ Administration Hospital, Columbia, MO, USA

    Gary L. Sieckman, Timothy J. Hoffman, Wynn A. Volkert & Leonard R. Forte

  4. Department of Internal Medicine, University of Missouri-Columbia, Columbia, MO, USA

    Timothy J. Hoffman

  5. Department of Medical Pharmacology and Physiology, University of Missouri-Columbia, Columbia, MO, USA

    Leonard R. Forte

  6. The Radiopharmaceutical Sciences Institute and Department of Radiology, University of Missouri-Columbia, 331 Hadley Hall, Columbia, MO, 65211, USA

    Michael F. Giblin

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  1. Michael F. Giblin
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  2. Hariprasad Gali
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  3. Gary L. Sieckman
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  4. Nellie K. Owen
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  5. Timothy J. Hoffman
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  6. Wynn A. Volkert
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  7. Leonard R. Forte
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Correspondence to Michael F. Giblin.

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Giblin, M.F., Gali, H., Sieckman, G.L. et al. In vitro and in vivo Evaluation of 111In-labeled E. coli Heat-Stable Enterotoxin Analogs for Specific Targeting of Human Breast Cancers. Breast Cancer Res Treat 98, 7–15 (2006). https://doi.org/10.1007/s10549-005-9040-8

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Key words

  • breast cancer
  • E. coli heat-stable enterotoxin
  • guanylate cyclase c
  • indium-111
  • uroguanylin