Identification, characterization and application of a new peptide against anterior gradient homolog 2 (AGR2)

Carolina Garri, Shannon Howell, Katrin Tiemann, Aleczandria Tiffany, Farzad Jalali-Yazdi, Mario M. Alba, Jonathan E. Katz, Terry T. Takahashi, Ralf Landgraf, Mitchell E. Gross, Richard W. Roberts, Kian Kani

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

The cancer-associated protein Anterior Gradient 2 (AGR2) has been described, predominantly in adenocarcinomas. Increased levels of extracellular AGR2 (eAGR2) have been correlated with poor prognosis in cancer patients, making it a potential biomarker. Additionally, neutralizing AGR2 antibodies showed preclinical effectiveness in murine cancer models suggesting eAGR2 may be a therapeutic target. We set out to identify a peptide by mRNA display that would serve as a theranostic tool targeting AGR2. This method enables the selection of peptides from a complex (> 1011) library and incorporates a protease incubation step that filters the selection for serum stable peptides. We performed six successive rounds of enrichment using a 10-amino acid mRNA display library and identified several AGR2 binding peptides. One of these peptides (H10), demonstrated high affinity binding to AGR2 with a binding constant (KD) of 6.4 nM. We developed an AGR2 ELISA with the H10 peptide as the capture reagent. Our H10-based ELISA detected eAGR2 from cancer cell spent media with a detection limit of (20-50 ng/ml). Furthermore, we investigated the therapeutic utility of H10 and discovered that it inhibited cell viability at IC50 (9-12 μmoles/L) in cancer cell lines. We also determined that 10 μg/ml of H10 was sufficient to inhibit cancer cell migration in breast and prostate cancer cell lines. A control peptide did not show any appreciable activity in these cells. The H10 peptide showed promise as both a novel diagnostic and a potential therapeutic peptide.

Original languageEnglish (US)
Pages (from-to)27363-27379
Number of pages17
JournalOncotarget
Volume9
Issue number44
DOIs
StatePublished - Jun 1 2018

Keywords

  • AGR2
  • Biomarker
  • Cancer
  • Therapeutic
  • mRNA display

ASJC Scopus subject areas

  • Oncology

Fingerprint Dive into the research topics of 'Identification, characterization and application of a new peptide against anterior gradient homolog 2 (AGR2)'. Together they form a unique fingerprint.

  • Cite this

    Garri, C., Howell, S., Tiemann, K., Tiffany, A., Jalali-Yazdi, F., Alba, M. M., Katz, J. E., Takahashi, T. T., Landgraf, R., Gross, M. E., Roberts, R. W., & Kani, K. (2018). Identification, characterization and application of a new peptide against anterior gradient homolog 2 (AGR2). Oncotarget, 9(44), 27363-27379. https://doi.org/10.18632/oncotarget.25221