Distinctive expression of circulating miR-22-3p and BCL11a as molecular markers of beta thalassemia

Thalassemia is a hereditary disease due to defective globin chain production that causes the accumulation of excessive globin (unbound) in the erythrocytes leading to ineffective erythropoiesis. Despite modern technologies, there is no ultimate treatment for thalassemia and most often blood transfusion is the likely management. Pakistan is among the countries with the highest thalassemia burden globally and ∼9000 newborns are suffering from thalassemia annually. The quest for molecular targets is of utmost importance and may be used for gene therapy of thalassemic patients. Circulating microRNAs (non-coding RNAs) have emerged as promising disease markers for diagnostic and therapeutic purposes. These small RNA molecules regulate the expression of genes throughout the human system including erythropoiesis and thalassemia. The present study was designed to explore the expression of miR-22-3p and its target gene BCL11a in plasma samples of beta-thalassemia patients. Plasma was isolated from the whole blood of a total of 100 participants including thalassemic patients as cases (n = 75) and healthy individuals as controls (n = 25). The relative expression of circulating miR-22-3p and BCL11a was measured in patients and controls using quantitative real-time PCR. The normalization was achieved using reference genes miR-16 and GAPDH for miR-22-3p and BCL11a, respectively. Normalized fold expression was calculated by using the 2− ΔΔCT method. The results showed significant up-regulation in circulating miR-22-3p expression in thalassemia patients when compared with controls (p < 0.05). However, BCL11a showed a significant down-regulation in the case of β-thalassemia in contrast to control individuals (p < 0.05). Moreover, it was observed that the expression level of miR-22-3p was not affected by age and gender differences (p > 0.05). Also, circulating levels of miR-22-3p were independent of clinical parameters such as Hb, RBCs, WBCs, HBA, HBA2, and HBF. Moreover, ROC analysis represented 86.67% sensitivity and 84% specificity of miR-22-3p with an AUC of 0.97. The sensitivity of BCL11a was 98.67% while the specificity of BCL11a was 100%. These results suggested that circulating miR-22-3p and its target BCL11a may be utilized as markers for the prediction of beta thalassemia. In future, these genetic markers may be utilized as therapeutic targets for the treatment of beta-thalassemia.