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Access Type
WSU Access
Date of Award
January 2024
Degree Type
Thesis
Degree Name
M.S.
Department
Pharmaceutical Sciences
First Advisor
ALOKE A. DUTTA
Abstract
Investigation of Inhibition of ferroptosis process by small molecules having antioxidant propertiesBy Vibha Deshpande July 2024
Advisor: Dr. Aloke DuttaMajor: Pharmaceutical Sciences Degree: Master of Science Introduction/ Objective Ferroptosis is a caspase-independent form of regulated cell death driven by iron-induced formation of lipid peroxides that accumulate at toxic levels causing cell death. Under physiological conditions, the cell combats lipid peroxidation with the help of selenoprotein GPX4. It has a central role in the ferroptosis pathway as it reduces lipid peroxide to lipid alcohol. Our study uses the utility of employing the ferroptosis-inducing compound RSL3 in cellular models to study the mechanism of production and inhibition of ferroptosis. Specifically, our investigation aims to study the molecules with potential antioxidant and iron-chelating properties for their capacity to inhibit the process of ferroptosis. Methods Our goal is to evaluate the effect of small molecules in modulating the ferroptosis pathway in cellular models. Test compounds are evaluated in different in vitro assays in PANC-1 cell lines sensitive to production of ferroptosis in presence of ferroptosis-inducing agent RSL3. The ability of test compounds to rescue cells from toxicity and oxidative stress upon treatment with RSL3 were evaluated. Various experiments like antiferroptotic cell viability study and DCFDA assays, , total GSH assay and study of different important biomarkers were carried out to judge the potential of compounds in modulating ferroptosis process and to provide antiferroptotic activity.
ResultsThe results show that dose dependent pretreatment of PANC-1 cells with multifunctional compounds D-512, D-583 and D-700 could protect the cells from toxicity of RSL-3 significantly compared to RSL-3 treated cells alone. The effect was found to be dose dependent. RSL-3 produces robust reactive oxygen species in PANC-1 cells. However, cells pre-treated with test compounds could dose dependently significantly reduce production of ROS upon exposure to RSL-3. The protection of level of GPX4 from exposure to RSL-3 by test compounds was evaluated. RSL3 reduces the level of GPX4. However, on pretreatment with the test compounds rescued the levels of GPX4. Similarly, SLC7A11 was rescued by pretreatment with the drug from toxic effects of RSL3. Another important biomarker for ferroptosis is NRF2. The loss of levels NRF2 upon treatment with RSL3 were restored by pretreatment with the test compounds. Finally, the levels of another marker FTH were unaltered after treatment with the test compounds. The test compound D-583 was able to protect the total GSH levels in the cells from toxic effect of RSL3 on treatment with our drugs. Conclusion In conclusion, the findings from this research present promising prospects for advancing innovative therapeutic approaches in the field of Parkinson's disease and other neurodegenerative disorders. Small molecules dopamine agonist with iron chelation and antioxidant properties can modulate ferroptosis process and offer a potential avenue to disease modifying therapeutics in PD. Drug D-512, D-583 and D-700 showed promising antiferroptotic activity in PANC-1 cells against the RSL3 induced toxicity.
Recommended Citation
Deshpande, Vibha, "Investigation Of Inhibition Of Ferroptosis Process By Small Molecules Having Antioxidant Properties" (2024). Wayne State University Theses. 958.
https://digitalcommons.wayne.edu/oa_theses/958