Transient Receptor Potential Ankyrin 1 (TRPA1) Modulation by 4-Hydroxynonenal (4-HNE) in Pancreatic Adenocarcinoma Cell Lines: Putative Roles for Therapies

Florentina Piciu; Dan Domocos; Gabriela Chiritoiu; Marioara Chiritoiu-Butnaru; Maria Mernea; Cezar Gabriel Popescu; Dragos Paul Mihai; Bianca Galateanu; Ariana Hudita; Alexandru Babes; Dana Cucu

doi:10.3390/ph17030344

Transient Receptor Potential Ankyrin 1 (TRPA1) Modulation by 4-Hydroxynonenal (4-HNE) in Pancreatic Adenocarcinoma Cell Lines: Putative Roles for Therapies

Pharmaceuticals (Basel). 2024 Mar 6;17(3):344. doi: 10.3390/ph17030344.

Authors

Florentina Piciu^{1

2}, Dan Domocos^{1

3}, Gabriela Chiritoiu⁴, Marioara Chiritoiu-Butnaru⁴, Maria Mernea¹, Cezar Gabriel Popescu¹, Dragos Paul Mihai⁵, Bianca Galateanu⁶, Ariana Hudita⁶, Alexandru Babes¹, Dana Cucu^{1

2}

Affiliations

¹ Department of Anatomy, Animal Physiology and Biophysics, Faculty of Biology, University of Bucharest, Spl. Independentei 91-95, 050095 Bucharest, Romania.
² Research Institute of the University of Bucharest (ICUB), University of Bucharest, 90-92 Sos. Panduri, 050663 Bucharest, Romania.
³ Cell Signalling Research Group, Institute of Biochemistry of the Romanian Academy, Splaiul Independenţei 296, 060031 Bucharest, Romania.
⁴ Department of Molecular and Cellular Biology, Institute of Biochemistry of the Romanian Academy, Splaiul Independentei 296, 060031 Bucharest, Romania.
⁵ Faculty of Pharmacy, "Carol Davila" University of Medicine and Pharmacy, 6 Traian Vuia Street, 020956 Bucharest, Romania.
⁶ Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Bucharest, Spl. Independentei 91-95, 050095 Bucharest, Romania.

Abstract

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.

Keywords: 4-HNE; PDAC; TRPA1; pancreatic adenocarcinoma.

Grants and funding

This work was supported by a grant from the Ministry of Research, Innovation and Digitization, CNCS-UEFISCDI, project number PN-III-P1-1.1-PD-2021-0076, within PNCDI III. The APC was funded by the University of Bucharest.