NERDG 2026
Poster 6 Abstract
Potential of L-Canavanine for Cancer Therapy
Kevin Ramirez Garcia (1), Mounika Pathuri (2), Raman Bahal (2), Anisha Gupta (1)
(1) School of Pharmacy and Physician Assistant Studies, University of Saint Joseph, CT, 06117 USA (2) School of Pharmacy, University of Connecticut, Storrs, CT, 06269 USA
Presenting Author: Kevin Ramirez Garcia
Corresponding Author: Anisha Gupta, [email protected]
Purpose
This study evaluates the cytotoxic and DNA-damaging effects of canavanine in human cancer cell lines. Double-strand breaks (DSBs) are particularly lethal forms of DNA damage, as they disrupt both DNA strands, impede replication and transcription, and often result in cell cycle arrest or apoptosis. Therapeutic strategies in oncology commonly exploit DSB induction, as observed with radiation and chemotherapeutic agents such as bleomycin, although their use is limited by toxicity and resistance. Natural products have long contributed to anticancer drug discovery, and attention has recently shifted to small molecules that modulate DNA repair or cause direct DNA damage. L-canavanine is a naturally occurring non-proteinogenic amino acid, structurally analogous to L-arginine. Owing to its ability to interfere with arginine-dependent cellular processes, L-canavanine has emerged as a promising anticancer candidate, particularly for tumors exhibiting arginine auxotrophy and high metabolic demand. This study highlights the therapeutic potential of L-canavanine in cancer treatment, focusing on its molecular mechanisms, selectivity toward malignant cells, and future translational prospects.
Methods
We used cell viability assays and γ-H2AX immunofluorescence to demonstrate that canavanine induces DSBs and selectively suppresses cancer cell growth, warranting further investigation. To evaluate the cytotoxic effects of L-Canavanine, A549, HeLa, and HepG2 cells were treated with increasing concentrations of the compound, and viability was measured using MTS and Cell Titer Glo assays. Both assays demonstrated a dose-dependent decrease in cell viability across all cell lines. DNA double-strand breaks were quantified using the γ-H2AX foci assay and Comet assay.
Results
Cell viability assays showed a dose-dependent decrease across cancer cell lines. Immunofluorescence microscopy revealed a robust γ-H2AX signal in L-Canavanine-treated A549 cells, comparable to that observed with the positive control bleomycin (150 µM), whereas untreated cells displayed minimal background staining. These results confirm that L-Canavanine is capable of inducing DNA double-strand breaks in A549 cells. In continuation of these studies, we are testing the effect of L-canavanine on various cancer cell lines.
Conclusion
These findings suggest a potential therapeutic role for L-Canavanine as a natural product-based DNA-damaging agent for cancer therapy.
Keywords
L-Canavanine, arginine metabolism, cancer therapy, DNA double-strand breaks, replication stress, apoptosis
Poster 6 Abstract
Potential of L-Canavanine for Cancer Therapy
Kevin Ramirez Garcia (1), Mounika Pathuri (2), Raman Bahal (2), Anisha Gupta (1)
(1) School of Pharmacy and Physician Assistant Studies, University of Saint Joseph, CT, 06117 USA (2) School of Pharmacy, University of Connecticut, Storrs, CT, 06269 USA
Presenting Author: Kevin Ramirez Garcia
Corresponding Author: Anisha Gupta, [email protected]
Purpose
This study evaluates the cytotoxic and DNA-damaging effects of canavanine in human cancer cell lines. Double-strand breaks (DSBs) are particularly lethal forms of DNA damage, as they disrupt both DNA strands, impede replication and transcription, and often result in cell cycle arrest or apoptosis. Therapeutic strategies in oncology commonly exploit DSB induction, as observed with radiation and chemotherapeutic agents such as bleomycin, although their use is limited by toxicity and resistance. Natural products have long contributed to anticancer drug discovery, and attention has recently shifted to small molecules that modulate DNA repair or cause direct DNA damage. L-canavanine is a naturally occurring non-proteinogenic amino acid, structurally analogous to L-arginine. Owing to its ability to interfere with arginine-dependent cellular processes, L-canavanine has emerged as a promising anticancer candidate, particularly for tumors exhibiting arginine auxotrophy and high metabolic demand. This study highlights the therapeutic potential of L-canavanine in cancer treatment, focusing on its molecular mechanisms, selectivity toward malignant cells, and future translational prospects.
Methods
We used cell viability assays and γ-H2AX immunofluorescence to demonstrate that canavanine induces DSBs and selectively suppresses cancer cell growth, warranting further investigation. To evaluate the cytotoxic effects of L-Canavanine, A549, HeLa, and HepG2 cells were treated with increasing concentrations of the compound, and viability was measured using MTS and Cell Titer Glo assays. Both assays demonstrated a dose-dependent decrease in cell viability across all cell lines. DNA double-strand breaks were quantified using the γ-H2AX foci assay and Comet assay.
Results
Cell viability assays showed a dose-dependent decrease across cancer cell lines. Immunofluorescence microscopy revealed a robust γ-H2AX signal in L-Canavanine-treated A549 cells, comparable to that observed with the positive control bleomycin (150 µM), whereas untreated cells displayed minimal background staining. These results confirm that L-Canavanine is capable of inducing DNA double-strand breaks in A549 cells. In continuation of these studies, we are testing the effect of L-canavanine on various cancer cell lines.
Conclusion
These findings suggest a potential therapeutic role for L-Canavanine as a natural product-based DNA-damaging agent for cancer therapy.
Keywords
L-Canavanine, arginine metabolism, cancer therapy, DNA double-strand breaks, replication stress, apoptosis
