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A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells
Published on: July 3, 2013
BacMam-enabled LanthaScreen cellular assays for PI3K/Akt pathway compound profiling in disease-relevant cell
Coby B Carlson1, Michael J Mashock, Kun Bi
1Invitrogen Discovery Assays and Services, Cell Systems Division, Invitrogen (Part of Life Technologies), Madison, Wisconsin 53719, USA. coby.carlson@invitrogen.com
Journal of Biomolecular Screening
|February 11, 2010
Summary
Researchers developed a new cell-based assay using BacMam technology to study the PI3K/Akt pathway. This method effectively profiles compounds in various cancer-relevant cell lines, aiding drug discovery.
Area of Science:
- Biochemistry
- Cell Biology
- Pharmacology
Background:
- The PI3K/Akt pathway is crucial for cell growth, survival, and cancer progression.
- There is a need for versatile cell-based assays for compound profiling in diverse disease contexts.
Purpose of the Study:
- To expand the LanthaScreen assay platform using BacMam gene delivery for broader application.
- To validate the utility of BacMam-enabled LanthaScreen assays for profiling inhibitors in disease-relevant cell lines.
Main Methods:
- Utilized BacMam gene delivery to adapt LanthaScreen assays across multiple cell types.
- Detected Akt-dependent PRAS40 phosphorylation at Thr246 in 10 PI3K/Akt-activated cell lines.
- Generated inhibitory profiles for 17 known pathway inhibitors.
Main Results:
- Successfully detected target phosphorylation in various cancer-associated cell lines.
- Validated the BacMam-enabled LanthaScreen assay for rapid compound profiling.
- Demonstrated that genetic alterations in PI3K/Akt signaling impact compound inhibitory profiles.
Conclusions:
- The BacMam-enabled LanthaScreen assay platform is effective for studying PI3K/Akt signaling in diverse cell backgrounds.
- This approach facilitates rapid compound profiling in disease-relevant cellular models.
- Understanding genetic alterations is key to interpreting compound efficacy in PI3K/Akt-driven diseases.
