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Monitoring AKT activity and targeting in live tissue and disease contexts using a real-time Akt-FRET biosensor mouse
James R W Conway1,2,3,4, Sean C Warren1,2,4, Young-Kyung Lee1,4
1Garvan Institute of Medical Research & The Kinghorn Cancer Centre, Sydney, NSW 2010, Australia.
Abstract:
Aberrant AKT activation occurs in a number of cancers, metabolic syndrome, and immune disorders, making it an important target for the treatment of many diseases. To monitor spatial and temporal AKT activity in a live setting, we generated an Akt-FRET biosensor mouse that allows longitudinal assessment of AKT activity using intravital imaging in conjunction with image stabilization and optical window technology. We demonstrate the sensitivity of the Akt-FRET biosensor mouse using various cancer models and verify its suitability to monitor response to drug targeting in spheroid and organotypic models. We also show that the dynamics of AKT activation can be monitored in real time in diverse tissues, including in individual islets of the pancreas, in the brown and white adipose tissue, and in the skeletal muscle. Thus, the Akt-FRET biosensor mouse provides an important tool to study AKT dynamics in live tissue contexts and has broad preclinical applications.
Insights
A novel Akt-FRET biosensor mouse enables real-time monitoring of AKT activity in live tissues. This tool aids in studying cancer, metabolic, and immune disorders, advancing preclinical research.
Area of Science:
- Biomedical research
- Molecular biology
- Cancer research
Background:
- Aberrant AKT signaling is implicated in various diseases including cancers, metabolic syndrome, and immune disorders.
- AKT is a critical therapeutic target for numerous conditions.
- Monitoring AKT activity in real-time is crucial for understanding disease mechanisms and treatment response.
Purpose of the Study:
- To develop a novel Akt-FRET biosensor mouse for longitudinal assessment of AKT activity in vivo.
- To validate the biosensor's sensitivity and utility in various disease models.
- To demonstrate real-time monitoring of AKT dynamics in diverse live tissues.
Main Methods:
- Generation of a genetically encoded Akt-FRET biosensor mouse.
- Intravital imaging with image stabilization and optical window technology.
- Application in cancer models, spheroid and organotypic cultures, and various live tissues (pancreas, adipose tissue, skeletal muscle).
Main Results:
- The Akt-FRET biosensor mouse allows sensitive, longitudinal monitoring of AKT activity.
- The biosensor is effective in various cancer models and suitable for assessing drug response.
- Real-time AKT dynamics were successfully visualized in pancreatic islets, adipose tissue, and skeletal muscle.
Conclusions:
- The Akt-FRET biosensor mouse is a valuable tool for studying AKT signaling dynamics in live tissue.
- This technology offers broad preclinical applications for understanding disease and evaluating therapeutic interventions.
- Enables real-time insights into AKT pathway behavior in complex biological systems.
