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Updated: Jul 13, 2025
SA-β-Galactosidase-Based Screening Assay for the Identification of Senotherapeutic Drugs
Published on: June 28, 2019
Genome-wide CRISPR activation screening in senescent cells reveals SOX5 as a driver and therapeutic target of
Yaobin Jing1, Xiaoyu Jiang2, Qianzhao Ji2
1State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China; Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, China; Beijing Institute for Stem Cell and Regenerative Medicine, Beijing 100101, China; School of Future Technology, University of Chinese Academy of Sciences, Beijing 100190, China.
Abstract:
Our understanding of the molecular basis for cellular senescence remains incomplete, limiting the development of strategies to ameliorate age-related pathologies by preventing stem cell senescence. Here, we performed a genome-wide CRISPR activation (CRISPRa) screening using a human mesenchymal precursor cell (hMPC) model of the progeroid syndrome. We evaluated targets whose activation antagonizes cellular senescence, among which SOX5 outperformed as a top hit. Through decoding the epigenomic landscapes remodeled by overexpressing SOX5, we uncovered its role in resetting the transcription network for geroprotective genes, including HMGB2. Mechanistically, SOX5 binding elevated the enhancer activity of HMGB2 with increased levels of H3K27ac and H3K4me1, raising HMGB2 expression so as to promote rejuvenation. Furthermore, gene therapy with lentiviruses carrying SOX5 or HMGB2 rejuvenated cartilage and alleviated osteoarthritis in aged mice. Our study generated a comprehensive list of rejuvenators, pinpointing SOX5 as a potent driver for rejuvenation both in vitro and in vivo.
Insights
SOX5 activation rejuvenates cells by resetting gene networks, promoting the expression of protective genes like HMGB2. Gene therapy with SOX5 or HMGB2 reversed aging in mice, offering new strategies for age-related diseases.
Area of Science:
- Molecular Biology
- Genetics
- Aging Research
Background:
- Cellular senescence, a key aging process, is not fully understood, hindering treatments for age-related diseases.
- Preventing stem cell senescence is crucial for ameliorating age-related pathologies.
Purpose of the Study:
- To identify genes that antagonize cellular senescence through genome-wide screening.
- To investigate the molecular mechanisms by which SOX5 promotes cellular rejuvenation.
Main Methods:
- Genome-wide CRISPR activation (CRISPRa) screening in human mesenchymal precursor cells (hMPCs).
- Epigenomic landscape analysis to understand SOX5's regulatory role.
- In vivo studies using gene therapy in aged mice to assess rejuvenation effects.
Main Results:
- SOX5 was identified as a top hit for antagonizing cellular senescence.
- SOX5 overexpression resets the transcription network of geroprotective genes, including HMGB2, by enhancing enhancer activity and histone modifications (H3K27ac, H3K4me1).
- Gene therapy with SOX5 or HMGB2 rejuvenated cartilage and alleviated osteoarthritis in aged mice.
Conclusions:
- SOX5 is a potent driver of cellular rejuvenation, both in vitro and in vivo.
- Targeting SOX5 and HMGB2 offers a promising therapeutic strategy for age-related diseases and tissue degeneration.
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