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

2 Citations (Scopus)

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

Fingerprint

Peptides
Neoplasms
Enzyme-Linked Immunosorbent Assay
Cell Line
Messenger RNA
Inhibitory Concentration 50
Libraries
Cell Movement
Limit of Detection
Prostatic Neoplasms
Cell Survival
Adenocarcinoma
Peptide Hydrolases
Therapeutics
Biomarkers
Breast Neoplasms
Amino Acids
Antibodies
Serum
Proteins

Keywords

  • AGR2
  • Biomarker
  • Cancer
  • mRNA display
  • Therapeutic

ASJC Scopus subject areas

  • Oncology

Cite this

Garri, C., Howell, S., Tiemann, K., Tiffany, A., Jalali-Yazdi, F., Alba, M. M., ... 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

Identification, characterization and application of a new peptide against anterior gradient homolog 2 (AGR2). / Garri, Carolina; Howell, Shannon; Tiemann, Katrin; Tiffany, Aleczandria; Jalali-Yazdi, Farzad; Alba, Mario M.; Katz, Jonathan E.; Takahashi, Terry T.; Landgraf, Ralf; Gross, Mitchell E.; Roberts, Richard W.; Kani, Kian.

In: Oncotarget, Vol. 9, No. 44, 01.06.2018, p. 27363-27379.

Research output: Contribution to journalArticle

Garri, C, Howell, S, Tiemann, K, Tiffany, A, Jalali-Yazdi, F, Alba, MM, Katz, JE, Takahashi, TT, Landgraf, R, Gross, ME, Roberts, RW & Kani, K 2018, 'Identification, characterization and application of a new peptide against anterior gradient homolog 2 (AGR2)', Oncotarget, vol. 9, no. 44, pp. 27363-27379. https://doi.org/10.18632/oncotarget.25221
Garri C, Howell S, Tiemann K, Tiffany A, Jalali-Yazdi F, Alba MM et al. Identification, characterization and application of a new peptide against anterior gradient homolog 2 (AGR2). Oncotarget. 2018 Jun 1;9(44):27363-27379. https://doi.org/10.18632/oncotarget.25221
Garri, Carolina ; Howell, Shannon ; Tiemann, Katrin ; Tiffany, Aleczandria ; Jalali-Yazdi, Farzad ; Alba, Mario M. ; Katz, Jonathan E. ; Takahashi, Terry T. ; Landgraf, Ralf ; Gross, Mitchell E. ; Roberts, Richard W. ; Kani, Kian. / Identification, characterization and application of a new peptide against anterior gradient homolog 2 (AGR2). In: Oncotarget. 2018 ; Vol. 9, No. 44. pp. 27363-27379.
@article{6fa51371050943a2bea6ee024a45b509,
title = "Identification, characterization and application of a new peptide against anterior gradient homolog 2 (AGR2)",
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.",
keywords = "AGR2, Biomarker, Cancer, mRNA display, Therapeutic",
author = "Carolina Garri and Shannon Howell and Katrin Tiemann and Aleczandria Tiffany and Farzad Jalali-Yazdi and Alba, {Mario M.} and Katz, {Jonathan E.} and Takahashi, {Terry T.} and Ralf Landgraf and Gross, {Mitchell E.} and Roberts, {Richard W.} and Kian Kani",
year = "2018",
month = "6",
day = "1",
doi = "10.18632/oncotarget.25221",
language = "English (US)",
volume = "9",
pages = "27363--27379",
journal = "Oncotarget",
issn = "1949-2553",
publisher = "Impact Journals",
number = "44",

}

TY - JOUR

T1 - Identification, characterization and application of a new peptide against anterior gradient homolog 2 (AGR2)

AU - Garri, Carolina

AU - Howell, Shannon

AU - Tiemann, Katrin

AU - Tiffany, Aleczandria

AU - Jalali-Yazdi, Farzad

AU - Alba, Mario M.

AU - Katz, Jonathan E.

AU - Takahashi, Terry T.

AU - Landgraf, Ralf

AU - Gross, Mitchell E.

AU - Roberts, Richard W.

AU - Kani, Kian

PY - 2018/6/1

Y1 - 2018/6/1

N2 - 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.

AB - 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.

KW - AGR2

KW - Biomarker

KW - Cancer

KW - mRNA display

KW - Therapeutic

UR - http://www.scopus.com/inward/record.url?scp=85048220445&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85048220445&partnerID=8YFLogxK

U2 - 10.18632/oncotarget.25221

DO - 10.18632/oncotarget.25221

M3 - Article

AN - SCOPUS:85048220445

VL - 9

SP - 27363

EP - 27379

JO - Oncotarget

JF - Oncotarget

SN - 1949-2553

IS - 44

ER -