Assessment of Myeloid Response and Iron Status among Sudanese Pediatric ESRD on Hemodialysis through Reticulocyte Parameters and β-Globin mRNA Expression

Abstract

Abstract Background: This study explores anemia in Sudanese children with End-Stage Renal Disease (ESRD) on hemodialysis, focusing on the effectiveness of Recombinant Human Erythropoietin (rHuEPO) treatment. It aims to assess myeloid response and classify anemia types using reticulocyte parameters and β-globin mRNA expression. Methods: Conducted in Khartoum State hospitals, the study included anemic children (≤15 years) undergoing hemodialysis and rHuEPO treatment. Flow Cytometry and RT-PCR were used to evaluate reticulocyte parameters and β-globin mRNA expression, respectively. Statistical analyses included Pearson correlation, logistic regression, and Receiver Operator Characteristics (ROC) curves. Results: This study evaluated β-globin gene expression in 45 anemic Sudanese children using RT-PCR, revealing variable expression levels ranging from 1.2 to 5.3 folds. Flow Cytometry showed significant correlations with mRNA Fold Change Expression (FCE). Gated Mature Reticulocytes and Immature Reticulocyte Fraction (IRF) positively correlated with FCE (p-values <0.0001 and 0.0258, respectively), whereas Hemoglobin Content of Reticulocytes (CHr) had a negative correlation (p = 0.0022). Gender analysis indicated a significant association between gender and IRF, with males showing lower IRF (p = 0.017), supported by area under the curve (AUC) analysis (AUC = 0.800). Flow Cytometry reticulocyte count correlated strongly with mRNA expression (β = 0.1018, p < 0.0001). There was no significant gender differences in hemoglobin levels (p = 0.999), but a strong positive correlation between hemoglobin and β-globin mRNA expression (r = 0.875, p < 0.000) was observed. Age was identified as a significant predictor of CHr (β = 0.394, p = 0.039). Conclusion: Significant variability in β-globin gene expression was observed. Flow Cytometry parameters showed strong correlations with mRNA expression. Gender differences influenced IRF levels, with males showing lower IRF. CHr and percentage of hypochromic red blood cells (%Hypo) were effective in assessing iron status and guiding rHuEPO dosage. The study highlights the potential of β-globin mRNA and reticulocyte parameters in diagnosing and monitoring anemia in pediatric ESRD patients. It underscores the need for precise, gender-specific approaches in treatment, laying a foundation for future research and improved therapeutic strategies.