An enhanced bioluminescence-based Annexin V probe for apoptosis detection in vitro and in vivo

The process of controlled cellular death known as apoptosis has an important central role not only in normal homeostatic maintenance of tissues, but also in numerous diseases such as cancer, neurodegenerative, autoimmune, and cardiovascular diseases. As a result, new technologies with the capability...

Full description

Saved in:
Bibliographic Details
Published in:Cell death & disease 2017-05, Vol.8 (5), p.e2826-e2826
Main Authors: Head, Trajen, Dau, Peter, Duffort, Stephanie, Daftarian, Pirouz, Joshi, Pratibha M, Vazquez-Padron, Roberto, Deo, Sapna K, Daunert, Sylvia
Format: Article
Language:English
Subjects:
Age
Animal models
Animals
Annexin A5 - metabolism
Annexin V
Apoptosis
Arteriosclerosis
Bioluminescence
Biosensors
Calorimetry
Cancer
Cardiovascular diseases
Cell death
Cornea
Data processing
Disease
Disease Models, Animal
Female
Fusion protein
Humans
Ischemia
Jurkat Cells
Luciferases, Renilla - metabolism
Luminescence
Macular degeneration
Mice, Inbred BALB C
Models, Biological
Molecular Probes - metabolism
Neurodegeneration
Original
Rats
Recombinant Fusion Proteins - metabolism
Reperfusion
Retina
Surgery
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The process of controlled cellular death known as apoptosis has an important central role not only in normal homeostatic maintenance of tissues, but also in numerous diseases such as cancer, neurodegenerative, autoimmune, and cardiovascular diseases. As a result, new technologies with the capability to selectively detect apoptotic cells represent a central focus of research for the study of these conditions. We have developed a new biosensor for the detection of apoptotic cells, incorporating the targeted selectivity for apoptotic cells from Annexin V with the sensitivity of bioluminescence signal generation from a serum-stable mutant of Renilla luciferase (RLuc8). Our data presents a complete characterization of the structural and biochemical properties of this new Annexin-Renilla fusion protein (ArFP) construct, as well as a validation of its ability to detect apoptosis in vitro. Moreover, this work represents the first report of a bioluminescent Annexin V apoptosis sensor utilized in vivo. With this new construct, we examine apoptosis within disease-relevant animal models of surgery-induced ischemia/reperfusion, corneal injury, and retinal cell death as a model of age-related macular degeneration. In each of these experiments, we demonstrate successful application of the ArFP construct for detection and bioluminescence imaging of apoptosis within each disease or treatment model. ArFP represents an important new tool in the continuously growing kit of technologies for apoptosis detection, and our results from both in vitro and in vivo experiments suggest a diverse range of potential clinically relevant applications including cancer therapeutic screening and efficacy analysis, atherosclerosis and cardiovascular disease detection, and the monitoring of any number of other conditions in which apoptosis has a central role.
ISSN:2041-4889
2041-4889
DOI:10.1038/cddis.2017.141