Integrated real-time imaging of executioner caspase dynamics, apoptosis-induced proliferation, and immunogenic cell death using a stable fluorescent reporter platform
Abstract Regulated cell death plays a central role in tissue homeostasis, disease progression, and therapeutic responses. However, tools to study these processes with high spatiotemporal resolution in physiologically relevant systems remain limited. Here, we present a fluorescent reporter cell syste...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Publishing Group
2025-08-01
|
| Series: | Cell Death Discovery |
| Online Access: | https://doi.org/10.1038/s41420-025-02662-y |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849767030412017664 |
|---|---|
| author | Selen Selcen Lena Wieland Tiago De Oliveira Michael Ghadimi Lena-Christin Conradi Günter Schneider Leonie Witte Matthias Wirth |
| author_facet | Selen Selcen Lena Wieland Tiago De Oliveira Michael Ghadimi Lena-Christin Conradi Günter Schneider Leonie Witte Matthias Wirth |
| author_sort | Selen Selcen |
| collection | DOAJ |
| description | Abstract Regulated cell death plays a central role in tissue homeostasis, disease progression, and therapeutic responses. However, tools to study these processes with high spatiotemporal resolution in physiologically relevant systems remain limited. Here, we present a fluorescent reporter cell system that enables real-time visualization of caspase-3/-7 activity via a DEVD-based biosensor, alongside a constitutive fluorescent marker for assessing successful transduction and cell presence. We generated stable cell lines expressing this reporter and adapted them to both 2D and 3D culture systems, including organoids. This platform allowed dynamic tracking of apoptotic events and viability loss at single-cell resolution. Using a proliferation dye, we also detected apoptosis-induced proliferation in neighboring cells. Furthermore, the system enabled simultaneous detection of immunogenic cell death via an endpoint measurement of surface calreticulin exposure by flow cytometry, supporting its application in studying immunogenic signaling. By measuring and integrating multiple cell death readouts by live-cell imaging, our system is well-suited for high-content screening and mechanistic dissection of different modes of cell death. When combined with complementary markers of pyroptosis and necroptosis, this platform may also be extended to investigate more complex, integrated forms of cell death. |
| format | Article |
| id | doaj-art-e86abfc0653c43ffb18f8f704b52c649 |
| institution | DOAJ |
| issn | 2058-7716 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Nature Publishing Group |
| record_format | Article |
| series | Cell Death Discovery |
| spelling | doaj-art-e86abfc0653c43ffb18f8f704b52c6492025-08-20T03:04:22ZengNature Publishing GroupCell Death Discovery2058-77162025-08-011111810.1038/s41420-025-02662-yIntegrated real-time imaging of executioner caspase dynamics, apoptosis-induced proliferation, and immunogenic cell death using a stable fluorescent reporter platformSelen Selcen0Lena Wieland1Tiago De Oliveira2Michael Ghadimi3Lena-Christin Conradi4Günter Schneider5Leonie Witte6Matthias Wirth7Department of General, Visceral and Pediatric Surgery, University Medical Center GöttingenDepartment of General, Visceral and Pediatric Surgery, University Medical Center GöttingenDepartment of General, Visceral and Pediatric Surgery, University Medical Center GöttingenDepartment of General, Visceral and Pediatric Surgery, University Medical Center GöttingenDepartment of General, Visceral and Pediatric Surgery, University Medical Center GöttingenDepartment of General, Visceral and Pediatric Surgery, University Medical Center GöttingenDepartment of General, Visceral and Pediatric Surgery, University Medical Center GöttingenDepartment of General, Visceral and Pediatric Surgery, University Medical Center GöttingenAbstract Regulated cell death plays a central role in tissue homeostasis, disease progression, and therapeutic responses. However, tools to study these processes with high spatiotemporal resolution in physiologically relevant systems remain limited. Here, we present a fluorescent reporter cell system that enables real-time visualization of caspase-3/-7 activity via a DEVD-based biosensor, alongside a constitutive fluorescent marker for assessing successful transduction and cell presence. We generated stable cell lines expressing this reporter and adapted them to both 2D and 3D culture systems, including organoids. This platform allowed dynamic tracking of apoptotic events and viability loss at single-cell resolution. Using a proliferation dye, we also detected apoptosis-induced proliferation in neighboring cells. Furthermore, the system enabled simultaneous detection of immunogenic cell death via an endpoint measurement of surface calreticulin exposure by flow cytometry, supporting its application in studying immunogenic signaling. By measuring and integrating multiple cell death readouts by live-cell imaging, our system is well-suited for high-content screening and mechanistic dissection of different modes of cell death. When combined with complementary markers of pyroptosis and necroptosis, this platform may also be extended to investigate more complex, integrated forms of cell death.https://doi.org/10.1038/s41420-025-02662-y |
| spellingShingle | Selen Selcen Lena Wieland Tiago De Oliveira Michael Ghadimi Lena-Christin Conradi Günter Schneider Leonie Witte Matthias Wirth Integrated real-time imaging of executioner caspase dynamics, apoptosis-induced proliferation, and immunogenic cell death using a stable fluorescent reporter platform Cell Death Discovery |
| title | Integrated real-time imaging of executioner caspase dynamics, apoptosis-induced proliferation, and immunogenic cell death using a stable fluorescent reporter platform |
| title_full | Integrated real-time imaging of executioner caspase dynamics, apoptosis-induced proliferation, and immunogenic cell death using a stable fluorescent reporter platform |
| title_fullStr | Integrated real-time imaging of executioner caspase dynamics, apoptosis-induced proliferation, and immunogenic cell death using a stable fluorescent reporter platform |
| title_full_unstemmed | Integrated real-time imaging of executioner caspase dynamics, apoptosis-induced proliferation, and immunogenic cell death using a stable fluorescent reporter platform |
| title_short | Integrated real-time imaging of executioner caspase dynamics, apoptosis-induced proliferation, and immunogenic cell death using a stable fluorescent reporter platform |
| title_sort | integrated real time imaging of executioner caspase dynamics apoptosis induced proliferation and immunogenic cell death using a stable fluorescent reporter platform |
| url | https://doi.org/10.1038/s41420-025-02662-y |
| work_keys_str_mv | AT selenselcen integratedrealtimeimagingofexecutionercaspasedynamicsapoptosisinducedproliferationandimmunogeniccelldeathusingastablefluorescentreporterplatform AT lenawieland integratedrealtimeimagingofexecutionercaspasedynamicsapoptosisinducedproliferationandimmunogeniccelldeathusingastablefluorescentreporterplatform AT tiagodeoliveira integratedrealtimeimagingofexecutionercaspasedynamicsapoptosisinducedproliferationandimmunogeniccelldeathusingastablefluorescentreporterplatform AT michaelghadimi integratedrealtimeimagingofexecutionercaspasedynamicsapoptosisinducedproliferationandimmunogeniccelldeathusingastablefluorescentreporterplatform AT lenachristinconradi integratedrealtimeimagingofexecutionercaspasedynamicsapoptosisinducedproliferationandimmunogeniccelldeathusingastablefluorescentreporterplatform AT gunterschneider integratedrealtimeimagingofexecutionercaspasedynamicsapoptosisinducedproliferationandimmunogeniccelldeathusingastablefluorescentreporterplatform AT leoniewitte integratedrealtimeimagingofexecutionercaspasedynamicsapoptosisinducedproliferationandimmunogeniccelldeathusingastablefluorescentreporterplatform AT matthiaswirth integratedrealtimeimagingofexecutionercaspasedynamicsapoptosisinducedproliferationandimmunogeniccelldeathusingastablefluorescentreporterplatform |