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Updated: Sep 22, 2025
An Optogenetic Method to Control and Analyze Gene Expression Patterns in Cell-to-cell Interactions
Published on: March 22, 2018
Optogenetic control of NOTCH1 signaling
Joanna Kałafut1, Jakub Czapiński1, Alicja Przybyszewska-Podstawka1
1Department of Biochemistry and Molecular Biology, Medical University of Lublin, 21-093, Lublin, Poland.
Abstract:
The Notch signaling pathway is a crucial regulator of cell differentiation as well as tissue organization, whose deregulation is linked to the pathogenesis of different diseases. NOTCH1 plays a key role in breast cancer progression by increasing proliferation, maintenance of cancer stem cells, and impairment of cell death. NOTCH1 is a mechanosensitive receptor, where mechanical force is required to activate the proteolytic cleavage and release of the Notch intracellular domain (NICD). We circumvent this limitation by regulating Notch activity by light. To achieve this, we have engineered an optogenetic NOTCH1 receptor (optoNotch) to control the activation of NOTCH1 intracellular domain (N1ICD) and its downstream transcriptional activities. Using optoNotch we confirm that NOTCH1 activation increases cell proliferation in MCF7 and MDA-MB-468 breast cancer cells in 2D and spheroid 3D cultures, although causing distinct cell-type specific migratory phenotypes. Additionally, optoNotch activation induced chemoresistance on the same cell lines. OptoNotch allows the fine-tuning, ligand-independent, regulation of N1ICD activity and thus a better understanding of the spatiotemporal complexity of Notch signaling. Video Abstract.
Insights
Scientists developed optoNotch, a light-activated system to control NOTCH1 signaling. This tool precisely regulates Notch signaling in breast cancer cells, revealing its role in proliferation, migration, and chemoresistance.
Area of Science:
- Cell Biology
- Molecular Biology
- Cancer Research
Background:
- The Notch signaling pathway regulates cell differentiation and tissue organization.
- Deregulation of Notch signaling is implicated in various diseases, including breast cancer.
- NOTCH1 promotes breast cancer progression by enhancing proliferation and cancer stem cell maintenance.
Purpose of the Study:
- To develop a light-inducible system (optoNotch) for precise control of NOTCH1 signaling.
- To investigate the role of NOTCH1 activation in breast cancer cell behavior and drug resistance.
- To understand the spatiotemporal dynamics of Notch signaling in cancer.
Main Methods:
- Engineering of an optogenetic NOTCH1 receptor (optoNotch).
- Activation of optoNotch using light to control Notch intracellular domain (N1ICD) activity.
- Assessment of NOTCH1 effects on cell proliferation, migration, and chemoresistance in MCF7 and MDA-MB-468 cells using 2D and 3D cultures.
Main Results:
- OptoNotch successfully enabled light-controlled, ligand-independent activation of N1ICD.
- NOTCH1 activation significantly increased proliferation in breast cancer cells.
- Distinct cell-type specific migratory phenotypes and induced chemoresistance were observed upon optoNotch activation.
Conclusions:
- OptoNotch provides a powerful tool for fine-tuning N1ICD activity with spatiotemporal control.
- This system enhances the understanding of Notch signaling complexity in breast cancer.
- OptoNotch activation contributes to increased proliferation, altered migration, and chemoresistance in breast cancer cells.
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