8,9-EET analogs protect glomerular cells from doxorubicin-induced toxicity

Introduction Doxorubicin (DOX) is an anthracycline chemotherapeutic whose clinical utility is limited by dose-dependent nephrotoxicity that contributes to acute and chronic kidney injury. Podocytes are essential components of the glomerular filtration barrier and are particularly vulnerable to DOX-induced oxidative stress, cytoskeletal disruption, loss of adhesion, and apoptosis. Therapeutic strategies that protect podocytes without compromising anticancer efficacy therefore represent an important unmet need. This study investigated the protective mechanisms and translational potential of 8,9-epoxyeicosatrienoic acid (8,9-EET) analogs against DOX-induced podocyte injury. Methods Nineteen 8,9-EET analogs were screened in cultured podocytes exposed to DOX. Protective effects were evaluated by assessing cell viability, caspase-3/7 activation, reactive oxygen species (ROS) generation, and focal adhesion integrity using confocal immunofluorescence analysis of vinculin. Selected analogs were also evaluated in DU145 prostate cancer and HCC1806 breast cancer cells to determine whether podocyte protection altered the anticancer activity of DOX. Results and Discussion Several 8,9-EET analogs significantly improved podocyte viability and attenuated DOX-induced apoptosis, as demonstrated by reduced caspase-3/7 activation. DOX markedly disrupted focal adhesion complexes, decreasing vinculin number and area, whereas co-treatment with selected analogs preserved vinculin-associated focal adhesion integrity. Effective analogs also significantly reduced DOX-induced ROS generation, consistent with attenuation of oxidative stress. Importantly, selected analogs protected podocytes without compromising DOX anticancer activity in either DU145 or HCC1806 cells, supporting separation of renoprotective effects from the desired cytotoxic action of DOX. These findings identify selected 8,9-EET analogs as promising podocyte-protective agents against DOX-induced injury. Their protective effects were associated with reduced oxidative stress and apoptosis and preservation of focal adhesion architecture, suggesting coordinated maintenance of podocyte structural integrity and survival. Preservation of DOX anticancer activity in prostate and breast cancer cells further supports the translational potential of these compounds as adjunctive therapies during anthracycline chemotherapy. Collectively, targeting epoxy lipid signaling with 8,9-EET analogs may provide a strategy to limit chemotherapy-associated podocyte injury and preserve renal function without diminishing antitumor efficacy. These results support preclinical evaluation of lead analogs, including their pharmacological properties, renal protective efficacy, and therapeutic compatibility with DOX in relevant models. These findings warrant defining efficacy, safety, dosing, and translational feasibility in vivo .