A fluorogenic cyclic peptide for imaging and quantification of drug-induced apoptosis

Nicole D. Barth; Ramon Subiros-Funosas; Lorena Mendive-Tapia; Rodger Duffin; Mario A. Shields; Jennifer A. Cartwright; Sónia Troeira Henriques; Jesus Sot; Felix M. Goñi; Rodolfo Lavilla; John A. Marwick; Sonja Vermeren; Adriano G. Rossi; Mikala Egeblad; Ian Dransfield; Marc Vendrell

doi:10.1038/s41467-020-17772-7

A fluorogenic cyclic peptide for imaging and quantification of drug-induced apoptosis

Nicole D. Barth, Ramon Subiros-Funosas, Lorena Mendive-Tapia, Rodger Duffin, Mario A. Shields, Jennifer A. Cartwright, Sónia Troeira Henriques, Jesus Sot, Felix M. Goñi, Rodolfo Lavilla, John A. Marwick, Sonja Vermeren, Adriano G. Rossi, Mikala Egeblad, Ian Dransfield^*, Marc Vendrell^*

^*Corresponding author for this work

Medicine, Hematology Oncology Division

Research output: Contribution to journal › Article › peer-review

59 Scopus citations

Abstract

Programmed cell death or apoptosis is a central biological process that is dysregulated in many diseases, including inflammatory conditions and cancer. The detection and quantification of apoptotic cells in vivo is hampered by the need for fixatives or washing steps for non-fluorogenic reagents, and by the low levels of free calcium in diseased tissues that restrict the use of annexins. In this manuscript, we report the rational design of a highly stable fluorogenic peptide (termed Apo-15) that selectively stains apoptotic cells in vitro and in vivo in a calcium-independent manner and under wash-free conditions. Furthermore, using a combination of chemical and biophysical methods, we identify phosphatidylserine as a molecular target of Apo-15. We demonstrate that Apo-15 can be used for the quantification and imaging of drug-induced apoptosis in preclinical mouse models, thus creating opportunities for assessing the in vivo efficacy of anti-inflammatory and anti-cancer therapeutics.

Original language	English (US)
Article number	4027
Journal	Nature communications
Volume	11
Issue number	1
DOIs	https://doi.org/10.1038/s41467-020-17772-7
State	Published - Dec 1 2020

Funding

N.D.B. acknowledges funding from OPTIMA (EP/L016559/1). R.S.F. acknowledges an MSCA Individual Fellowship (659046). L.M.T. acknowledges the support of Fundacion Antonio Martin Escudero (FAME) in the form of a post-doctoral fellowship. S.T.H. is an Australian Research Council Future Fellow (FT150100398). R.L. acknowledges funding from MINECO (CTQ2015-67870-P and ERA NET PCIN-2015-224). F.M.G. acknowledges financial help from the Spanish Ministry of Economy (FEDER MINECO PGC2018-099857-B-I00) and the Basque Government (IT264-19). M.E. acknowledges funding from Cold Spring Harbor Laboratory Cancer Center (P30-CA045508), North-well Health, and the Thompson Family Foundation. M.V. acknowledges funding from an ERC Consolidator Grant (771443), the Biotechnology and Biological Sciences Research Council (BB/M025160/1), and The Royal Society (RG160289). The authors thank the technical support from the QMRI Flow Cytometry and Confocal Advanced Light Microscopy facilities at the University of Edinburgh and Yaiza Varela (UPV), Luxembourg Bio Technologies Ltd. (Rehovot) for the kind supply of reagents for peptide synthesis, and Astex Therapeutics, who kindly provided AT7519 as a gift. The authors also thank Prof. Chris Gregory (University of Edinburgh) for provision of BL-2 cells and valuable discussions.

ASJC Scopus subject areas

General Chemistry
General Biochemistry, Genetics and Molecular Biology
General Physics and Astronomy

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1038/s41467-020-17772-7

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Barth, N. D., Subiros-Funosas, R., Mendive-Tapia, L., Duffin, R., Shields, M. A., Cartwright, J. A., Henriques, S. T., Sot, J., Goñi, F. M., Lavilla, R., Marwick, J. A., Vermeren, S., Rossi, A. G., Egeblad, M., Dransfield, I., & Vendrell, M. (2020). A fluorogenic cyclic peptide for imaging and quantification of drug-induced apoptosis. Nature communications, 11(1), Article 4027. https://doi.org/10.1038/s41467-020-17772-7

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title = "A fluorogenic cyclic peptide for imaging and quantification of drug-induced apoptosis",

abstract = "Programmed cell death or apoptosis is a central biological process that is dysregulated in many diseases, including inflammatory conditions and cancer. The detection and quantification of apoptotic cells in vivo is hampered by the need for fixatives or washing steps for non-fluorogenic reagents, and by the low levels of free calcium in diseased tissues that restrict the use of annexins. In this manuscript, we report the rational design of a highly stable fluorogenic peptide (termed Apo-15) that selectively stains apoptotic cells in vitro and in vivo in a calcium-independent manner and under wash-free conditions. Furthermore, using a combination of chemical and biophysical methods, we identify phosphatidylserine as a molecular target of Apo-15. We demonstrate that Apo-15 can be used for the quantification and imaging of drug-induced apoptosis in preclinical mouse models, thus creating opportunities for assessing the in vivo efficacy of anti-inflammatory and anti-cancer therapeutics.",

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Barth, ND, Subiros-Funosas, R, Mendive-Tapia, L, Duffin, R, Shields, MA, Cartwright, JA, Henriques, ST, Sot, J, Goñi, FM, Lavilla, R, Marwick, JA, Vermeren, S, Rossi, AG, Egeblad, M, Dransfield, I & Vendrell, M 2020, 'A fluorogenic cyclic peptide for imaging and quantification of drug-induced apoptosis', Nature communications, vol. 11, no. 1, 4027. https://doi.org/10.1038/s41467-020-17772-7

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AU - Barth, Nicole D.

AU - Subiros-Funosas, Ramon

AU - Mendive-Tapia, Lorena

AU - Duffin, Rodger

AU - Shields, Mario A.

AU - Cartwright, Jennifer A.

AU - Henriques, Sónia Troeira

AU - Sot, Jesus

AU - Goñi, Felix M.

AU - Lavilla, Rodolfo

AU - Marwick, John A.

AU - Vermeren, Sonja

AU - Rossi, Adriano G.

AU - Egeblad, Mikala

AU - Dransfield, Ian

AU - Vendrell, Marc

PY - 2020/12/1

Y1 - 2020/12/1

N2 - Programmed cell death or apoptosis is a central biological process that is dysregulated in many diseases, including inflammatory conditions and cancer. The detection and quantification of apoptotic cells in vivo is hampered by the need for fixatives or washing steps for non-fluorogenic reagents, and by the low levels of free calcium in diseased tissues that restrict the use of annexins. In this manuscript, we report the rational design of a highly stable fluorogenic peptide (termed Apo-15) that selectively stains apoptotic cells in vitro and in vivo in a calcium-independent manner and under wash-free conditions. Furthermore, using a combination of chemical and biophysical methods, we identify phosphatidylserine as a molecular target of Apo-15. We demonstrate that Apo-15 can be used for the quantification and imaging of drug-induced apoptosis in preclinical mouse models, thus creating opportunities for assessing the in vivo efficacy of anti-inflammatory and anti-cancer therapeutics.

AB - Programmed cell death or apoptosis is a central biological process that is dysregulated in many diseases, including inflammatory conditions and cancer. The detection and quantification of apoptotic cells in vivo is hampered by the need for fixatives or washing steps for non-fluorogenic reagents, and by the low levels of free calcium in diseased tissues that restrict the use of annexins. In this manuscript, we report the rational design of a highly stable fluorogenic peptide (termed Apo-15) that selectively stains apoptotic cells in vitro and in vivo in a calcium-independent manner and under wash-free conditions. Furthermore, using a combination of chemical and biophysical methods, we identify phosphatidylserine as a molecular target of Apo-15. We demonstrate that Apo-15 can be used for the quantification and imaging of drug-induced apoptosis in preclinical mouse models, thus creating opportunities for assessing the in vivo efficacy of anti-inflammatory and anti-cancer therapeutics.

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A fluorogenic cyclic peptide for imaging and quantification of drug-induced apoptosis

Abstract

Funding

ASJC Scopus subject areas

UN SDGs

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