Canonical DDR activation by EMT inducing agent 5-Fluorouracil is modulated by a cannabinoid based combinatorial
Khalid Bashir Mir1, Souneek Chakraborty2, Tanzeeba Amin1
1Academy of Scientific & Innovative Research (AcSIR), Ghaziabad, India; Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Jammu 180001, India.
Abstract:
Anastasis cascade including induction of Epithelial to Mesenchymal Transition (EMT), DNA repair, and stimulation of pro-survival mediators collectively exaggerate therapy resistance in cancer prognosis. The extensive implications of DNA-damaging agents are clinically proven futile for the rapid development of disease recurrence during treatment regime. Herein we report a glycosidic derivative of Δ9-tetrahydrocannabinol (THC-9-OG) abrogates sub-toxic doses of 5-Fluorouracil (5FU) induced EMT in colon cancer cells nullifying DNA repairing mechanism. Our in vitro and in vivo data strongly proclaims that THC-9-OG could not only abrogate 5FU mediated background EMT activation through stalling matrix degradation as well as murine 4T1 lung metastasis but also vigorously diminished Rad-51 repairing mediator along with stimulation of γ-H2AX foci formation. The combinatorial treatment (5FU + THC-9-OG) in Apc knockout colorectal carcinoma model conferred remission of the crypt progenitor phenotype which was prominently identified in 5FU treatment. Mechanistically, we demonstrated that 5FU plus THC-9-OG significantly attenuated major EMT inducer Vimentin via extensive ROS generation along with autophagy induction via LC3B I-II conversion and p62 degradation in a p-ATM dependent manner. Additionally, Cannabinoid receptor CB1 was responsible for abrogation of Vimentin since we found increase in the expression of γH2AX and decrease in vimentin expression in CB1 agonist (ACEA) plus 5FU treated cells. Nutshell, our results unveil a new direction of Cannabinoid based combinatorial approach to control background EMT along with robust enhancing of DNA damage potential of sub-toxic concentration of 5FU resulting immense inhibition of distant metastasis coupled with triggering cell death in vitro and in vivo.
Insights
A novel cannabinoid derivative, THC-9-OG, combined with 5-Fluorouracil (5FU), overcomes cancer therapy resistance by blocking DNA repair and inhibiting Epithelial to Mesenchymal Transition (EMT). This combination halts metastasis and promotes cancer cell death.
Area of Science:
- Oncology
- Cancer Therapeutics
- Molecular Biology
Background:
- Therapy resistance in cancer is exacerbated by Epithelial to Mesenchymal Transition (EMT), DNA repair mechanisms, and pro-survival signaling.
- DNA-damaging chemotherapy agents often prove ineffective due to rapid disease recurrence.
- Targeting EMT and DNA repair pathways is crucial for improving cancer treatment outcomes.
Purpose of the Study:
- To investigate the efficacy of a novel glycosidic derivative of Δ9-tetrahydrocannabinol (THC-9-OG) in combination with 5-Fluorouracil (5FU).
- To determine if THC-9-OG can abrogate 5FU-induced EMT and enhance DNA damage in colon cancer cells.
- To elucidate the underlying molecular mechanisms of the combinatorial therapy.
Main Methods:
- In vitro studies using colon cancer cells and in vivo studies using murine models (4T1 lung metastasis and Apc knockout colorectal carcinoma model).
- Assessment of EMT markers (Vimentin), DNA repair proteins (Rad-51, γ-H2AX), autophagy markers (LC3B, p62), and reactive oxygen species (ROS) generation.
- Evaluation of Cannabinoid receptor CB1 involvement using a CB1 agonist (ACEA).
Main Results:
- THC-9-OG abrogated 5FU-induced EMT, stalled matrix degradation, and inhibited lung metastasis.
- The combination significantly diminished Rad-51, stimulated γ-H2AX foci, attenuated Vimentin expression via ROS and autophagy induction (LC3B I-II, p62 degradation) in a p-ATM dependent manner.
- Cannabinoid receptor CB1 activation was responsible for Vimentin downregulation and enhanced DNA damage.
Conclusions:
- The combination of 5FU and THC-9-OG represents a novel strategy to overcome cancer therapy resistance.
- This approach effectively controls background EMT, enhances DNA damage, inhibits metastasis, and induces cell death.
- Cannabinoid-based combinatorial therapies hold significant promise for improving colorectal cancer treatment.
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