A fluorophore-conjugated reagent enabling rapid detection, isolation and live tracking of senescent cells

Sophia Magkouta; Dimitris Veroutis; Athanasios Pousias; Angelos Papaspyropoulos; Natassa Pippa; Nikolaos Lougiakis; Konstantinos Kambas; Nefeli Lagopati; Aikaterini Polyzou; Maria Georgiou; Maria Chountoulesi; Stergios Pispas; Spyros Foutadakis; Nicole Pouli; Panagiotis Marakos; Athanassios Kotsinas; Panayotis Verginis; Dimitrios Valakos; Athanasia Mizi; Argyris Papantonis; Giannis Vatsellas; Panagiotis Galanos; Jiri Bartek; Russell Petty; Manuel Serrano; Dimitris Thanos; Charis Roussos; Marco Demaria; Konstantinos Evangelou; Vassilis G Gorgoulis

doi:10.1016/j.molcel.2023.09.006

A fluorophore-conjugated reagent enabling rapid detection, isolation and live tracking of senescent cells

Mol Cell. 2023 Oct 5;83(19):3558-3573.e7. doi: 10.1016/j.molcel.2023.09.006.

Authors

Sophia Magkouta¹, Dimitris Veroutis², Athanasios Pousias³, Angelos Papaspyropoulos², Natassa Pippa⁴, Nikolaos Lougiakis³, Konstantinos Kambas⁵, Nefeli Lagopati⁶, Aikaterini Polyzou⁷, Maria Georgiou³, Maria Chountoulesi⁸, Stergios Pispas⁹, Spyros Foutadakis¹⁰, Nicole Pouli³, Panagiotis Marakos³, Athanassios Kotsinas⁷, Panayotis Verginis¹¹, Dimitrios Valakos¹⁰, Athanasia Mizi¹², Argyris Papantonis¹², Giannis Vatsellas¹³, Panagiotis Galanos¹⁴, Jiri Bartek¹⁵, Russell Petty¹⁶, Manuel Serrano¹⁷, Dimitris Thanos¹⁸, Charis Roussos¹⁹, Marco Demaria²⁰, Konstantinos Evangelou⁷, Vassilis G Gorgoulis²¹

Affiliations

¹ Molecular Carcinogenesis Group, Department of Histology and Embryology, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece; Marianthi Simou and G.P.Livanos Labs, 1st Department of Critical Care and Pulmonary Services, School of Medicine, National & Kapodistrian University of Athens, "Evangelismos" Hospital, Athens, 10676, Greece.
² Molecular Carcinogenesis Group, Department of Histology and Embryology, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece; Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece.
³ Department of Pharmacy, Division of Pharmaceutical Chemistry, National and Kapodistrian University of Athens, Panepistimiopolis, Zografou, 15771 Athens, Greece.
⁴ Section of Pharmaceutical Technology, Department of Pharmacy, School of Health Sciences, National and Kapodistrian University of Athens, Panepistimioupolis Zografou, 15771 Athens, Greece; Theoretical and Physical Chemistry Institute, National Hellenic Research Foundation, 11635 Athens, Greece.
⁵ Hellenic Pasteur Institute, 11521 Athens, Greece.
⁶ Molecular Carcinogenesis Group, Department of Histology and Embryology, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece; Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece; Laboratory of Biology, Department of Basic Medical Sciences, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece.
⁷ Molecular Carcinogenesis Group, Department of Histology and Embryology, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece.
⁸ Section of Pharmaceutical Technology, Department of Pharmacy, School of Health Sciences, National and Kapodistrian University of Athens, Panepistimioupolis Zografou, 15771 Athens, Greece.
⁹ Theoretical and Physical Chemistry Institute, National Hellenic Research Foundation, 11635 Athens, Greece.
¹⁰ Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece.
¹¹ Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece; Laboratory of Immune Regulation and Tolerance, Division of Basic Sciences, University of Crete Medical School, 70013 Heraklion, Greece; Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology, 70013 Heraklion, Greece.
¹² Institute of Pathology, University Medical Center Göttingen, 37075 Göttingen, Germany; Clinical Research Unit 5002, University Medical Center Goettingen, 37075 Goettingen, Germany.
¹³ Greek Genome Center, Biomedical Research Foundation, Academy of Athens, 11527, Athens, Greece.
¹⁴ Genome Integrity Group, Danish Cancer Society Research Center, 2100 Copenhagen, Denmark.
¹⁵ Genome Integrity Group, Danish Cancer Society Research Center, 2100 Copenhagen, Denmark; Science for Life Laboratory, Division of Genome Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Solna, 171 77 Stockholm, Sweden.
¹⁶ Ninewells Hospital and Medical School, University of Dundee, DD19SY Dundee, UK.
¹⁷ Institute for Research in Biomedicine (IRB Barcelona), Barcelona Institute of Science and Technology (BIST), 08028 Barcelona, Spain; Altos Labs, Cambridge Institute of Science, Granta Park CB21 6GP, United Kingdom.
¹⁸ Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece; Greek Genome Center, Biomedical Research Foundation, Academy of Athens, 11527, Athens, Greece.
¹⁹ Marianthi Simou and G.P.Livanos Labs, 1st Department of Critical Care and Pulmonary Services, School of Medicine, National & Kapodistrian University of Athens, "Evangelismos" Hospital, Athens, 10676, Greece.
²⁰ European Research Institute for the Biology of Ageing (ERIBA), University Medical Center Groningen, 9713 AV Groningen, The Netherlands.
²¹ Molecular Carcinogenesis Group, Department of Histology and Embryology, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece; Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece; Ninewells Hospital and Medical School, University of Dundee, DD19SY Dundee, UK; Faculty Institute for Cancer Sciences, Manchester Academic Health Sciences Centre, University of Manchester, M20 4GJ Manchester, UK; Faculty of Health and Medical Sciences, University of Surrey, GU2 7YH Surrey, UK. Electronic address: vgorg@med.uoa.gr.

PMID: 37802028
DOI: 10.1016/j.molcel.2023.09.006

Abstract

Cellular senescence is a stress-response mechanism implicated in various physiological processes, diseases, and aging. Current detection approaches have partially addressed the issue of senescent cell identification in clinical specimens. Effective methodologies enabling precise isolation or live tracking of senescent cells are still lacking. In-depth analysis of truly senescent cells is, therefore, an extremely challenging task. We report (1) the synthesis and validation of a fluorophore-conjugated, Sudan Black-B analog (GLF16), suitable for in vivo and in vitro analysis of senescence by fluorescence microscopy and flow cytometry and (2) the development and application of a GLF16-carrying micelle vector facilitating GLF16 uptake by living senescent cells in vivo and in vitro. The compound and the applied methodology render isolation of senescent cells an easy, rapid, and precise process. Straightforward nanocarrier-mediated GLF16 delivery in live senescent cells comprises a unique tool for characterization of senescence at an unprecedented depth.

Keywords: GLF-16; flow cytometry; fluorescence microscopy; live tracking; micelle; optimized SBB analogs; senescence.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cellular Senescence*
Flow Cytometry
Indicators and Reagents

Substances

Indicators and Reagents