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Optimal cannabinoid-terpene combination ratios suppress mutagenicity of gastric reflux in normal and metaplastic esophageal cells
Aaron Goldman, Gabriel Gonzalez, Svetlana A Karpova, Leutz Buon, Masood A Shammas, Hiroshi Mashimo, Markus H Frank, Natasha Y Frank
Aaron Goldman, Department of Medicine, Brigham and Women’s Hospital, Boston, MA 02115, United States
Gabriel Gonzalez, Svetlana A Karpova, Natasha Y Frank, Department of Medicine, VA Boston Healthcare, Boston, MA 02132, United States
Gabriel Gonzalez, Svetlana A Karpova, Natasha Y Frank, Division of Genetics, Department of Medicine, Brigham and Women’s Hospital, Boston, MA 02115, United States
Leutz Buon, Department of Medicine, Dana Farber Harvard Cancer Institute, Boston, MA 02115, United States
Masood A Shammas, Department of Medical Oncology, Dana Farber Cancer Institute, Boston, MA 02132, United States
Hiroshi Mashimo, Gastroenterology Section, VA Boston Healthcare System, Harvard Medical School, Boston, MA 02130, United States
Markus H Frank, Transplant Research Program, Boston Children’s Hospital, Boston, MA 02115, United States
Markus H Frank, Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02139, United States
Co-corresponding authors: Aaron Goldman and Natasha Y Frank.
Author contributions: Goldman A and Gonzalez G designed and performed in vitro experiments and analyzed data; Karpova SA, Shammas MA, and Buon L assisted with experiments; Mashimo H, Frank MH and Frank NY supervised the project; Goldman A, Gonzalez G and Frank NY wrote and revised the paper; all authors reviewed and approved the final manuscript.
Supported by the National Institutes of Health, No. R01EY025794, No. R01HL161087, No. P01AG071463 and No. 1K01CA226375-01A1; and Veterans Administration Research and Development Merit Review Awards, No. 1I01BX000516, No. 1I01RX000989 and No. 1I01BX006004.
Institutional review board statement: As indicated in the revised manuscript, all tissue samples used for microarray analysis and tissue culture were obtained with patient consent from the respective institutions.
Conflict-of-interest statement: Goldman A, Gonzalez G, and Frank NY are inventors of a United States patent assigned to Brigham and Women’s Hospital and the VA Boston Healthcare System, Boston, MA, United States; Frank MH and Frank NY are inventors or co-inventors of United States and international patents assigned to Brigham and Women’s Hospital, Boston Children’s Hospital, the Massachusetts Eye and Ear Infirmary, and the VA Boston Healthcare System, Boston, MA, United States, licensed to Rheacell GmbH and CoKG (Heidelberg, Germany); Frank MH holds equity in and serves as a scientific advisor to Rheacell GmbH and Co. KG.
Data sharing statement: The datasets generated during and/or analyzed in the current study are available from the corresponding author upon reasonable request.
Corresponding author: Natasha Y Frank, MD, Department of Medicine, VA Boston Healthcare, 1400 VFW Parkway, Building 3, Suite 2A105, Boston, MA 02132, United States.
nyfrank@bwh.harvard.edu
Received: July 1, 2025
Revised: September 12, 2025
Accepted: February 4, 2026
Published online: April 14, 2026
Processing time: 276 Days and 15.3 Hours
BACKGROUND
Esophageal adenocarcinoma (EAC) frequently arises from chronic exposure to acid and bile reflux, with secondary bile acids, such as deoxycholic acid (DCA), contributing to its pathogenesis through mechanisms involving reactive oxygen species (ROS), oxidative DNA damage, and resistance to apoptosis. The human endocannabinoid system (ECS) regulates diverse anti-inflammatory, antioxidant, and analgesic pathways implicated in disease modulation and may counteract the harmful effects of gastroesophageal reflux. Despite its therapeutic promise, effective pharmacological activation of the ECS remains challenging.
AIM
To evaluate whether specific cannabinoid-terpene combinations targeting the ECS could attenuate the mutagenic and cytotoxic effects of bile acid-induced stress in esophageal cell models, and to assess the clinical significance of ECS-related protein receptors in the progression of EAC.
METHODS
Human esophageal epithelial cells exposed to DCA, as well as a Barrett’s esophagus cell line subjected to low potential of hydrogen and a bile acid cocktail, were treated with various ratios of phyto-cannabinoids and terpenes. Endpoints included DNA damage, mitochondrial membrane potential, and ROS production to identify optimal compound combinations. Expression of ECS-related receptor proteins in clinical samples was assessed by immunohistochemistry.
RESULTS
A 1:5 ratio of cannabigerol to phytol significantly reduced DCA-induced DNA damage, preserved mitochondrial membrane potential, and decreased ROS levels. This combination also promoted apoptosis in damaged cells and reduced mutagenicity. Analysis of patient samples demonstrated that expression of the ECS-associated receptor protein CB1 correlated with EAC progression, indicating a broader clinical role for ECS modulation in cancer prevention.
CONCLUSION
Modulation of the ECS using systematically selected cannabinoid-terpene ratios can attenuate bile acid-induced esophageal damage and may reduce carcinogenic progression. These findings support further in vivo studies and raise the possibility of expanding cannabinoid-terpene therapeutics to other diseases with comparable pathogenic mechanisms.
Core Tip: Activation of the endocannabinoid system (ECS) with specific cannabinoid-terpene combinations and ratios has been shown to protect esophageal cells from bile acid-induced DNA damage and mutations, which are key drivers of esophageal adenocarcinoma (EAC). In vitro models revealed that a 1:5 ratio of cannabigerol to phytol or β-caryophyllene significantly reduced DNA damage and oxidative stress while preserving mitochondrial function. This combination also promoted apoptosis in damaged cells, while the ECS receptor expression patterns correlated with EAC progression. These findings indicate that ECS modulation may represent a novel therapeutic strategy for EAC and related conditions.
