Background: The cardiotoxic effects of doxorubicin (DOX), a powerful chemotherapeutic drug, are widely recognized. Cuproptosis, a unique copper-dependent form of controlled cell death, may be involved in DOX-induced cardiomyopathy, according to recent findings. This study employs both in vivo and in silico procedures to investigate the protective effects of the copper chelator penicillamine (PEN) and the role of cuproptosis in DOX-related cardiotoxicity. Methods: Thirty-two adult Sprague Dawley rats were allocated into four groups (n = 8): control, DOX, DOX+PEN, and PEN. Cardiac function was assessed via echocardiography. Serum cardiac biomarkers (LDH, CK-MB, CTnI), oxidative stress markers (SOD, GPX, MDA), and expression levels of cuproptosis-related genes (FDX1, LIAS, SLC31A1, ATP7A) were evaluated. Histopathological examinations and immunohistochemical staining for FDX1, SLC31A1, and DLAT were performed. Molecular docking simulated PEN’s interaction with cuproptosis-related proteins. Network pharmacology and molecular docking studies were also conducted to identify core molecular targets and simulate PEN’s binding interactions with key cuproptosis regulators. Results: DOX administration induced significant cardiac dysfunction, oxidative stress, and upregulation of cuproptosis markers. PEN treatment mitigated these effects, improved cardiac function, reduced fibrosis, and suppressed the expression of cuproptosis-related genes and proteins. Docking results confirmed strong interactions between PEN and cuproptosis-regulatory proteins. Network pharmacology revealed 14 key overlapping targets linking PEN with cuproptosis and DOX-induced cardiotoxicity. Conclusion: This study provides experimental evidence implicating cuproptosis in DOX-induced cardiomyopathy. PEN exerts cardioprotection, potentially by targeting this pathway, offering a promising therapeutic strategy.

Background: Differentiated thyroid carcinoma (DTC) is the most common malignant tumor of the endocrine system. Our study is aimed to investigate the role of the α- and γ-Klotho genes in the development and progression of DTC. Methods: The expression of the α- and γ-Klotho genes was assessed by quantitative real-time polymerase chain reaction (RT-qPCR) in thyroid tissues of 40 DTC patients and 40 age- and sex-matched subjects diagnosed as goiter and included as a control group. The demographic, laboratory and clinicopathological data of the subjects were reviewed to detect their association with the Klotho genes. Results: The α-Klotho gene expression was statistically significantly lower in DTC tissues compared to goiter tissues ( p < 0.001). However, there was no statistically significant association between the γ-Klotho gene expression and DTC ( p = 0.643). Receiver operating characteristic (ROC) curve analysis showed the diagnostic value of the α-Klotho mRNA levels, by providing an AUC of 0.954 value (95% CI = 0.910–0.999; p < 0.001). Lower α-Klotho gene expression associated significantly with higher stages of DTC ( p = 0.026). Logistic regression analysis declared that low α-Klotho mRNA expression was demonstrated to be a significant predictor for the likelihood of DTC on top of goiter ( p = 0.001). Conclusions: Our study showed the role of the α-Klotho gene as a potential novel biomarker for discrimination between DTC and goiter tissues. Low α-Klotho mRNA expression was found to be a significant predictor for the likelihood of DTC on top of goiter, as well as higher stages of this tumor.