Assessment of myeloid response and iron status among Sudanese pediatric ESKD on hemodialysis through reticulocyte parameters and β-globin mRNA expression

Abstract

Background: Sudanese children with End-Stage Kidney Disease (ESKD) often show limited improvement in hemoglobin levels despite treatment with recombinant human erythropoietin (rHuEPO). This study aims to assess the response to rHuEPO therapy by analyzing β-globin mRNA expression and reticulocyte parameters. Additionally, it classifies anemia among Sudanese pediatric patients based on iron status, considering age and gender as biological markers for evaluating treatment response.

Methods: A prospective observational cohort study was conducted from January 2019 to February 2020 in Khartoum, Sudan, involving 45 anemic children aged 2 to 15 years diagnosed with ESKD. The treatment protocol included rHuEPO injections and maintenance hemodialysis. Laboratory assessments consisted of complete blood count (CBC), absolute reticulocyte count, ferritin, and transferrin measurements. β-globin mRNA expression was quantified using reverse transcription polymerase chain reaction (RT-PCR), and reticulocyte parameters, including Reticulocyte Hemoglobin Content (CHr), percentage of hypochromic reticulocytes (HYPO%), and Immature Reticulocyte Fraction (IRF), were measured via flow cytometry.

Results: Significant variations in hemoglobin levels were observed across different age groups (p = 0.011). Gender analysis revealed a significant association with IRF, showing a lower IRF in male patients (p = 0.017). However, there were no significant differences in hemoglobin levels between genders (p = 0.999). β-globin mRNA expression showed considerable variability, with a strong positive correlation with hemoglobin levels (r = 0.875, p < 0.0001).

Conclusion: Age and gender significantly influence treatment responses in children with ESKD, highlighting the need to consider growth physiology in anemia management. This study underscores the variability in β-globin mRNA expression and its association with Flow Cytometry parameters, demonstrating their effectiveness in evaluating iron status and guiding rHuEPO dosage.

Keywords: End-stage kidney disease (ESKD); Ferritin; Flow cytometry; Hypochromic cells (HYPO%); Immature reticulocyte fraction (IRF); Reticulocyte hemoglobin content (CHr); Sudanese children; β-globin mRNA expression.

Fig. 1

Comparison between Flow Cytometry and Manual Reticulocyte Counts in β-globin mRNA Expression

Fig. 2

Estimated Marginal Means of Hemoglobin among Anemic Sudanese Children with ESKD

Fig. 3

β-globin expression fold changes among studied patients

Fig. 4

The area under curve ROC curve expression of anemic Sudanese gender effect on IRF

Fig. 5

CHr shows an inverse correlation with MCV among the studied anemic Sudanese children with ESKD

Fig. 6

Significant correlation between patient’s age and CHr among the studied anemic Sudanese children with ESKD

Fig. 7

Positive insignificant correlation between Hb and CHr among the studied anemic Sudanese children with ESKD

Fig. 8

Positive insignificant correlation between MCHC and CHr among the studied anemic Sudanese children with ESKD

Fig. 9

Positive insignificant correlation between Ferritin and CHr among the studied anemic Sudanese children with ESKD

Fig. 10

Negative insignificant correlation between Transferrin and CHr among the studied anemic Sudanese children with ESKD

Fig. 11

The ROC curve of CHr, mRNA, and Hb versus gender among the studied anemic Sudanese children with ESKD

Fig. 12

The ROC curve of CHr, mRNA, Hb versus Hypo% among the studied anemic Sudanese children with ESKD