Multi-Omics Analysis of Red Blood Cells Reveals Molecular Pathways Underlying Thalassemia Severity Beyond Globin Gene Mutations

BackgroundHemoglobinopathies are the most common monogenic genetic disorders, primarily managed through blood transfusions or bone marrow transplantation. Clinical severity other than mutational effect not well investigated and still unknown. This study aims to identify dysregulated molecular pathways in red blood cells contributing to thalassemia severity. MethodFrom a cohort of 285 hemoglobinopathy patients, 10 age-matched individuals with identical compound heterozygous mutations (IVS 1-5 G>C and CD 26 G>A) were screened. Five had severe thalassemia requiring regular transfusions, while five had a non-severe form requiring fewer transfusions. RNA sequencing and proteome analysis were conducted on isolated RBCs, through Novaseq and Orbitrap MS platform respectively. Bioconductor-R and different bioinformatics tools were utilized subsequently. ResultsTranscriptome analysis revealed an increased percentage of snRNA transcripts in all over thalassemia group. Pathways related to autophagy, mitophagy, and chaperone-mediated folding were enriched in the severe group. Thus, dysregulated genes, linked to ineffective erythropoiesis were fished out also. ConclusionIn this this study, thalassemia subjects were of same mutational genotype, but clinically opposite severity. Accordingly, first time identified six pathways which are responsible for thalassemia severity independent of mutational burden. These dysregulated pathways can be further be explored and targeted experimentally for drug development.