Transient Receptor Potential Ankyrin 1 (TRPA1) Modulation by 4-Hydroxynonenal (4-HNE) in Pancreatic Adenocarcinoma
Florentina Piciu1,2, Dan Domocos1,3, Gabriela Chiritoiu4
1Department of Anatomy, Animal Physiology and Biophysics, Faculty of Biology, University of Bucharest, Spl. Independentei 91-95, 050095 Bucharest, Romania.
Background:
Transient receptor potential channels (TRP) are overexpressed in some pancreatic adenocarcinoma (PDAC) patients and cell lines, settling them as putative therapeutic targets in this disease. Reactive oxygen species (ROS), with levels increased in PDAC, modulate some members of the TRP family renamed "redox channels". Here, we investigate the direct effects of 4-hydroxinonenal (4-HNE) on TRPA1, natively expressed in PDAC cell lines and in association with cell migration and cell cycle progression.
Methods:
We performed microfluorimetry experiments, while the activation of resident membrane channels was investigated using confocal microscopy. We applied a prospective molecular docking of 4-HNE using Autodock and AutoDock Tools4. Also, we simulated the diffusion of 4-HNE through the membrane from the extracellular space with the Permeability of Molecules across Membranes (PerMM) web server. The analysis of cell migration was performed using the wound healing assay, and cell cycle progression was acquired using a Beckman Coulter CytoFlex flow cytometer.
Results:
Our results show, for the first time in PDAC, that 4-HNE diffuses through the cell membrane and rapidly activates Ca2+ uptake in PDAC cells. This process depends on TRPA1 activation, as 4-HNE forms a covalent binding with a pocket-like region within the intracellular N-terminal of the channel, shaped by the cysteine residues 621, 641, and 665. The activation of TRPA1 by 4-HNE inhibits cell migration and induces cell cycle arrest in the G2/M phase.
Conclusions:
Our study brings new insights into the effects of 4-HNE, highlighting the activation of the TRPA1 channel, a druggable, putative target for PDAC-expressing tumors.
Insights
4-hydroxynonenal (4-HNE) activates the TRPA1 channel in pancreatic cancer cells, inhibiting migration and cell cycle progression. This finding identifies TRPA1 as a potential therapeutic target for pancreatic adenocarcinoma (PDAC).
Area of Science:
- Molecular Biology
- Cell Biology
- Cancer Research
Background:
- Pancreatic adenocarcinoma (PDAC) exhibits overexpression of Transient Receptor Potential (TRP) channels, making them potential therapeutic targets.
- Increased reactive oxygen species (ROS) in PDAC modulate TRP channels, termed "redox channels".
Purpose of the Study:
- To investigate the direct effects of 4-hydroxynonenal (4-HNE) on TRPA1 channels in PDAC.
- To determine the role of 4-HNE-TRPA1 interaction in PDAC cell migration and cell cycle progression.
Main Methods:
- Microfluorimetry and confocal microscopy to study channel activation.
- Molecular docking and membrane diffusion simulations to analyze 4-HNE interaction.
- Wound healing assay and flow cytometry to assess cell migration and cell cycle.
Main Results:
- 4-HNE rapidly diffuses across the cell membrane and activates Ca2+ uptake in PDAC cells via TRPA1.
- 4-HNE covalently binds to TRPA1 at cysteine residues 621, 641, and 665.
- TRPA1 activation by 4-HNE inhibits cell migration and induces G2/M phase cell cycle arrest.
Conclusions:
- 4-HNE activates TRPA1 channels in PDAC, impacting cell behavior.
- TRPA1 is a druggable target for pancreatic adenocarcinoma.
- This study provides novel insights into 4-HNE's role in PDAC progression.
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