Related Experiment Video
Updated: Sep 11, 2025
siRNA Screening to Identify Ubiquitin and Ubiquitin-like System Regulators of Biological Pathways in Cultured Mammalian Cells
Published on: May 24, 2014
Identification of novel SIRT1 up-regulators using a cell-based high-throughput screening assay.
Yanjia Shen1, Huilin Yang1, Peng Gao2
1Beijing Key Laboratory of Drug Targets Identification and Drug Screening, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, China.
Researchers developed a high-throughput screening assay to find FDA-approved drugs that boost Sirtuin 1 (SIRT1) expression. They identified 17 compounds, including HDAC inhibitors, that increase SIRT1, offering potential new treatments for neurodegenerative diseases.
Area of Science:
- Biochemistry
- Pharmacology
- Neuroscience
Background:
- Sirtuin 1 (SIRT1) is a key target for treating neurodegenerative disorders.
- Resveratrol is known to increase SIRT1 expression and activity.
- Identifying existing drugs to modulate SIRT1 offers a therapeutic repositioning strategy.
Purpose of the Study:
- To develop and validate a cell-based high-throughput screening (HTS) assay for identifying SIRT1 transcriptional up-regulators.
- To screen FDA-approved drugs for their ability to increase SIRT1 transcription.
- To evaluate potential drug candidates for neurodegenerative disease treatment.
Main Methods:
- Established a stable 293A cell line with a SIRT1-promoter-luciferase reporter construct.
- Utilized a 96-well microplate format for HTS, with resveratrol as a positive control.
- Screened 1523 FDA-approved drugs and performed dose-response and molecular docking analyses.
Main Results:
- The HTS assay demonstrated high reliability (Z' = 0.67).
- 17 out of 1523 screened drugs significantly upregulated SIRT1 transcription (>200%).
- Top compounds, including HDAC inhibitors (belinostat, panobinostat, vorinostat), enhanced SIRT1 mRNA and protein levels and interacted with SIRT1.
- Belinostat showed efficacy in an animal model of multiple sclerosis (EAE mice).
Conclusions:
- The developed HTS assay is effective for identifying SIRT1 up-regulators.
- Several FDA-approved drugs, particularly HDAC inhibitors, can significantly increase SIRT1 transcription and expression.
- These identified compounds represent promising candidates for drug repositioning in neurodegenerative diseases.
