Evaluating miRNAs as predictive biomarkers for drug-induced autoimmune toxicity in D-penicillamine induced autoimmune diseases animal model.

Abstract

Drug-induced autoimmune (DIA) toxicity represents a significant challenge in drug safety evaluation, and traditional autoantibody biomarker antinuclear antibodies (ANA) exhibits limitations in DIA assessment, highlighting the urgent need for more reliable biomarkers. This study employed d-penicillamine to induce an autoimmune disease model in Brown Norway (BN) rats and systematically investigated the expression changes of autoimmunity-related microRNAs (miRNAs) in spleen, exploring their potential value as novel biomarkers for predicting DIA toxicity. A total of 30 male BN rats were randomly divided into control group (0 mg/kg), low-dose d-penicillamine group (150 mg/kg), and high-dose group (450 mg/kg), with necropsy performed at days 7 and 14 post-administration. ANA detection, histopathological examination of spleen, surface immune-related proteins CD11a and CD278 on splenic CD3CD4T-cells, and miRNA expression profiling in spleen tissue were conducted. Receiver operating characteristic (ROC) curves were plotted to evaluate the correlation of ANA and miRNAs with the degree of splenic pathological damage, and the relationships between miRNAs and both ANA and CD11a/CD278 were analyzed. The results demonstrated that 16 miRNAs were significantly up-regulated in spleen tissue, among which 13 miRNAs showed superior predictive performance (AUC = 0.828-0.898) compared to the traditional biomarker ANA (AUC = 0.783), with most of these miRNAs exhibiting correlation with CD11a/CD278 expression. These findings indicate that aberrant miRNA expression in the d-penicillamine-induced autoimmune disease preclinical model in BN rats holds potential for early prediction of DIA toxicity, with some miRNAs demonstrating higher diagnostic value than ANA, thereby providing novel molecular biomarker candidates for early diagnosis of DIA toxicity.