A Molecular Signature for Anastasis, Recovery from the Brink of Apoptotic Cell Death

Fung

2018

JoVE

Anastasis (Greek for “rising to life”) is a recently discovered cell recovery phenomenon whereby dying cells can reverse late-stage cell death processes that are generally assumed to be intrinsically irreversible. Promoting anastasis could in principle rescue or preserve injured cells that are difficult to replace such as cardiomyocytes or neurons, thereby facilitating tissue recovery. Conversely, suppressing anastasis in cancer cells, undergoing apoptosis after anti-cancer therapies, may ensure cancer cell death and reduce the chances of recurrence. However, these studies have been hampered by the lack of tools for tracking the fate of cells that undergo anastasis in live animals. The challenge is to identify the cells that have reversed the cell death process despite their morphologically normal appearance after recovery. To overcome this difficulty, we have developed Drosophila and mammalian CaspaseTracker biosensor systems that can identify and permanently track the anastatic cells in vitro or in vivo. Here, we present in vivo protocols for the generation and use of the CaspaseTracker dual biosensor system to detect and track anastasis in Drosophila melanogaster after transient exposure to cell death stimuli. While conventional biosensors and protocols can label cells actively undergoing apoptotic cell death, the CaspaseTracker biosensor can permanently label cells that have recovered after caspase activation - a hallmark of late-stage apoptosis, and simultaneously identify active apoptotic processes. This biosensor can also track the recovery of the cells that attempted other forms of cell death that directly or indirectly involved caspase activity. Therefore, this protocol enables us to continuously track the fate of these cells and their progeny, facilitating future studies of the biological functions, molecular mechanisms, physiological and pathological consequences, and therapeutic implications of anastasis. We also discuss the appropriate controls to distinguish cells that undergo anastasis from those that display non-apoptotic caspase activity in vivo.

Section: Discussionsupporting

confidence: 80%

Detecting Anastasis <em>In Vivo</em> by CaspaseTracker Biosensor

Fung

2018

JoVE

“…Whole‐transcriptome RNA sequencing (RNAseq) was performed on untreated cells, ethanol‐treated apoptotic cells, and anastatic cells . Based on this study, the time course of anastasis can be divided into two stages: the earlier (before 4 h) and the later (4 h to 12 h) stages, both of which were different from the dying or untreated groups.…”

Section: Apoptosismentioning

confidence: 99%

To the edge of cell death and back

et al. 2018

Programmed cell death plays a central role in maintaining homeostasis. Various studies have demonstrated that programmed cell death is not a one‐way street; cells can survive even when the core cell death processes are underway. Cell death initiation, prevention, and recovery function in a coordinated fashion to establish and maintain a homeostatic environment. In this review, we discuss how dying cells can be rescued from death's grip and the subsequent physiological consequences. We suggest a fundamental question to be answered—at least at the single cell level is, can we predict if a certain cell is more or less likely to survive or die? And importantly, what physiological and pathological consequences, as well as therapeutic approaches can we delineate from this ability to predict cell death versus survival.

Section: What Is Anastasis?mentioning

confidence: 99%

“…Recovery from caspase activation after exposure to multiple types of stimuli has been observed. These include: 1) chemical apoptosis inducers such as ethanol, DMSO, staurosporine, jasplakinolide, and cucurbitacin [ 1 , 9 ]; 2) the death-inducing ligand TNFα combined with cycloheximide [ 9 ]; 3) physical inducers such as cold shock [ 10 ]; 4) physiological stress such as protein starvation [ 10 ]. Further study will be needed to uncover the diversity of apoptotic inducers from which cells can recover.…”

Section: What Kinds Of Apoptotic Stimuli Permit Anastasis?mentioning

confidence: 99%

Q&A: Cellular near death experiences—what is anastasis?

Sun

Montell

2017

BMC Biol

Self Cite

Apoptosis is a form of programmed cell death that is carried out by proteolytic enzymes called caspases. Executioner caspase activity causes cells to shrink, bleb, and disintegrate into apoptotic bodies and has been considered a point of no return for apoptotic cells. However, relatively recent work has shown that cells can survive transient apoptotic stimuli, even after executioner caspase activation. This process is called anastasis. In this Q&A, we answer common questions that arise regarding anastasis, including how it is defined, the origin of the name, the potential physiological consequences, molecular mechanisms, and open questions for this new field of study.