Discovery of a phosphatidylserine-recognizing peptide and its utility in molecular imaging of tumour apoptosis

Narendra Thapa, Soyoun Kim, In Sup So, Byung Heon Lee, Ick Chan Kwon, Kuiwon Choi, In-San Kim

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

The exposure of phosphatidylserine (PS) molecules from the inner to the outer leaflet of the plasma membrane has been recognized as a well-defined molecular epitope of cells undergoing apoptosis. Examination and monitoring of PS exposure is an extensively used molecular marker in non-invasive apoptosis imaging under a variety of clinical conditions, including the assessment of therapeutic anti-cancer agents and myocardial infarction. Herein, we report the identification of a PS-recognizing peptide which was identified by the screening of an M13 phage display peptide library onto PS-coated ELISA plates. Repeated biopanning for a total of four rounds revealed a predominant enrichment of the phage clone displaying peptide sequence, CLSYYPSYC (46%). The identified phage clone evidenced enhanced binding to a number of apoptotic cells over non-apoptotic cells, and this binding was inhibited by both annexin V and synthesized peptide displayed on the phage. The binding of the fluorescein-labelled CLSYYPSYC peptide to apoptotic versus normal cells was assessed by both FACS analysis and fluorescence microscopy. Optical imaging after the systemic administration of fluorescein-labelled CLSYYPSYC peptide to tumour-bearing nude mice (H460 cells xenograft model) treated with a single dose of an anticancer drug (camp-tothecin) indicated peptide homing to the tumour. The histological examination of tumour tissues showed intense staining of the tumour vasculature and apoptotic tumour cells. With these results, the CLSYYPSYC peptide is recognized as a novel PS-recognizing moiety which may possibly be developed into a molecular probe for the imaging of apoptosis in vivo. This application would clearly be relevant to assessments of the efficacy of anticancer therapy in tumours.

Original languageEnglish
Pages (from-to)1649-1660
Number of pages12
JournalJournal of Cellular and Molecular Medicine
Volume12
Issue number5A
DOIs
Publication statusPublished - 2008 Sep 1
Externally publishedYes

Fingerprint

Molecular Imaging
Phosphatidylserines
Apoptosis
Peptides
Neoplasms
Bacteriophages
Fluorescein
Clone Cells
Bacteriophage M13
Peptide Library
Molecular Probes
Annexin A5
Optical Imaging
Fluorescence Microscopy
Heterografts
Nude Mice
Epitopes
Cell Count
Enzyme-Linked Immunosorbent Assay
Myocardial Infarction

Keywords

  • Annexin V
  • Apoptosis
  • Peptide
  • Phage display
  • Phosphatidylserine

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Medicine

Cite this

Discovery of a phosphatidylserine-recognizing peptide and its utility in molecular imaging of tumour apoptosis. / Thapa, Narendra; Kim, Soyoun; So, In Sup; Lee, Byung Heon; Kwon, Ick Chan; Choi, Kuiwon; Kim, In-San.

In: Journal of Cellular and Molecular Medicine, Vol. 12, No. 5A, 01.09.2008, p. 1649-1660.

Research output: Contribution to journalArticle

Thapa, Narendra ; Kim, Soyoun ; So, In Sup ; Lee, Byung Heon ; Kwon, Ick Chan ; Choi, Kuiwon ; Kim, In-San. / Discovery of a phosphatidylserine-recognizing peptide and its utility in molecular imaging of tumour apoptosis. In: Journal of Cellular and Molecular Medicine. 2008 ; Vol. 12, No. 5A. pp. 1649-1660.
@article{5b0ac1e2a8d14f2a8e838ef34c874339,
title = "Discovery of a phosphatidylserine-recognizing peptide and its utility in molecular imaging of tumour apoptosis",
abstract = "The exposure of phosphatidylserine (PS) molecules from the inner to the outer leaflet of the plasma membrane has been recognized as a well-defined molecular epitope of cells undergoing apoptosis. Examination and monitoring of PS exposure is an extensively used molecular marker in non-invasive apoptosis imaging under a variety of clinical conditions, including the assessment of therapeutic anti-cancer agents and myocardial infarction. Herein, we report the identification of a PS-recognizing peptide which was identified by the screening of an M13 phage display peptide library onto PS-coated ELISA plates. Repeated biopanning for a total of four rounds revealed a predominant enrichment of the phage clone displaying peptide sequence, CLSYYPSYC (46{\%}). The identified phage clone evidenced enhanced binding to a number of apoptotic cells over non-apoptotic cells, and this binding was inhibited by both annexin V and synthesized peptide displayed on the phage. The binding of the fluorescein-labelled CLSYYPSYC peptide to apoptotic versus normal cells was assessed by both FACS analysis and fluorescence microscopy. Optical imaging after the systemic administration of fluorescein-labelled CLSYYPSYC peptide to tumour-bearing nude mice (H460 cells xenograft model) treated with a single dose of an anticancer drug (camp-tothecin) indicated peptide homing to the tumour. The histological examination of tumour tissues showed intense staining of the tumour vasculature and apoptotic tumour cells. With these results, the CLSYYPSYC peptide is recognized as a novel PS-recognizing moiety which may possibly be developed into a molecular probe for the imaging of apoptosis in vivo. This application would clearly be relevant to assessments of the efficacy of anticancer therapy in tumours.",
keywords = "Annexin V, Apoptosis, Peptide, Phage display, Phosphatidylserine",
author = "Narendra Thapa and Soyoun Kim and So, {In Sup} and Lee, {Byung Heon} and Kwon, {Ick Chan} and Kuiwon Choi and In-San Kim",
year = "2008",
month = "9",
day = "1",
doi = "10.1111/j.1582-4934.2008.00305.x",
language = "English",
volume = "12",
pages = "1649--1660",
journal = "Journal of Cellular and Molecular Medicine",
issn = "1582-1838",
publisher = "Wiley-Blackwell",
number = "5A",

}

TY - JOUR

T1 - Discovery of a phosphatidylserine-recognizing peptide and its utility in molecular imaging of tumour apoptosis

AU - Thapa, Narendra

AU - Kim, Soyoun

AU - So, In Sup

AU - Lee, Byung Heon

AU - Kwon, Ick Chan

AU - Choi, Kuiwon

AU - Kim, In-San

PY - 2008/9/1

Y1 - 2008/9/1

N2 - The exposure of phosphatidylserine (PS) molecules from the inner to the outer leaflet of the plasma membrane has been recognized as a well-defined molecular epitope of cells undergoing apoptosis. Examination and monitoring of PS exposure is an extensively used molecular marker in non-invasive apoptosis imaging under a variety of clinical conditions, including the assessment of therapeutic anti-cancer agents and myocardial infarction. Herein, we report the identification of a PS-recognizing peptide which was identified by the screening of an M13 phage display peptide library onto PS-coated ELISA plates. Repeated biopanning for a total of four rounds revealed a predominant enrichment of the phage clone displaying peptide sequence, CLSYYPSYC (46%). The identified phage clone evidenced enhanced binding to a number of apoptotic cells over non-apoptotic cells, and this binding was inhibited by both annexin V and synthesized peptide displayed on the phage. The binding of the fluorescein-labelled CLSYYPSYC peptide to apoptotic versus normal cells was assessed by both FACS analysis and fluorescence microscopy. Optical imaging after the systemic administration of fluorescein-labelled CLSYYPSYC peptide to tumour-bearing nude mice (H460 cells xenograft model) treated with a single dose of an anticancer drug (camp-tothecin) indicated peptide homing to the tumour. The histological examination of tumour tissues showed intense staining of the tumour vasculature and apoptotic tumour cells. With these results, the CLSYYPSYC peptide is recognized as a novel PS-recognizing moiety which may possibly be developed into a molecular probe for the imaging of apoptosis in vivo. This application would clearly be relevant to assessments of the efficacy of anticancer therapy in tumours.

AB - The exposure of phosphatidylserine (PS) molecules from the inner to the outer leaflet of the plasma membrane has been recognized as a well-defined molecular epitope of cells undergoing apoptosis. Examination and monitoring of PS exposure is an extensively used molecular marker in non-invasive apoptosis imaging under a variety of clinical conditions, including the assessment of therapeutic anti-cancer agents and myocardial infarction. Herein, we report the identification of a PS-recognizing peptide which was identified by the screening of an M13 phage display peptide library onto PS-coated ELISA plates. Repeated biopanning for a total of four rounds revealed a predominant enrichment of the phage clone displaying peptide sequence, CLSYYPSYC (46%). The identified phage clone evidenced enhanced binding to a number of apoptotic cells over non-apoptotic cells, and this binding was inhibited by both annexin V and synthesized peptide displayed on the phage. The binding of the fluorescein-labelled CLSYYPSYC peptide to apoptotic versus normal cells was assessed by both FACS analysis and fluorescence microscopy. Optical imaging after the systemic administration of fluorescein-labelled CLSYYPSYC peptide to tumour-bearing nude mice (H460 cells xenograft model) treated with a single dose of an anticancer drug (camp-tothecin) indicated peptide homing to the tumour. The histological examination of tumour tissues showed intense staining of the tumour vasculature and apoptotic tumour cells. With these results, the CLSYYPSYC peptide is recognized as a novel PS-recognizing moiety which may possibly be developed into a molecular probe for the imaging of apoptosis in vivo. This application would clearly be relevant to assessments of the efficacy of anticancer therapy in tumours.

KW - Annexin V

KW - Apoptosis

KW - Peptide

KW - Phage display

KW - Phosphatidylserine

UR - http://www.scopus.com/inward/record.url?scp=54049124091&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=54049124091&partnerID=8YFLogxK

U2 - 10.1111/j.1582-4934.2008.00305.x

DO - 10.1111/j.1582-4934.2008.00305.x

M3 - Article

VL - 12

SP - 1649

EP - 1660

JO - Journal of Cellular and Molecular Medicine

JF - Journal of Cellular and Molecular Medicine

SN - 1582-1838

IS - 5A

ER -