title: Correction of aberrant pre-mRNA splicing by antisense oligonucleotides in beta-thalassemia Egyptian patients with IVSI-110 mutation.
abstract: The splicing mutation in intron 1 of beta-globin gene (IVS1-110) is the most common mutation in Egyptian thalassemics that causes aberrant splicing of pre-mRNA and deficient beta-globin chain synthesis. Antisense oligonucleotides (ASONs) are compounds that redirect pre-mRNA splicing and modify gene expression. Our aim was ex vivo correction of the aberrant splicing of beta-globin110 pre-mRNA by ASON against the 3' aberrant splice site. Peripheral blood mononuclear cells of 10 thalassemic patients with IVS1-110 mutation were duplicated and 1 was treated with 20 micromoL/mL morpholino ASON targeted against the 3' aberrant splice site. The level of total hemoglobin (Hb), fetal Hb, and mRNA were estimated in the duplicate samples. Five cases (50%) showed correction with ASON treatment, of which 2 cases showed the appearance of corrected mRNA band with absence of the aberrant band and 3 cases showed an increased ratio of the corrected to the aberrant mRNA band from 2:1 to 3:1, and 4:1. The total Hb showed significant increase in the 5 corrected cases. In conclusion, ASON can restore correct splicing of beta-globin pre-mRNA leading to correct gene product in cultured erythropoietic cells. These results suggest the applicability of ASON for the treatment of thalassemia.
authors.affiliation: Pediatric Department, Faculty of Medicine, Cairo University, Cairo, Egypt.
authors.forename: Amal
authors.initials: A
authors.lastname: El-Beshlawy
authors.forename: Azza
authors.initials: A
authors.lastname: Mostafa
authors.forename: Ilham
authors.initials: I
authors.lastname: Youssry
authors.forename: Hala
authors.initials: H
authors.lastname: Gabr
authors.forename: Iman M
authors.initials: IM
authors.lastname: Mansour
authors.forename: Manar
authors.initials: M
authors.lastname: El-Tablawy
authors.forename: Mona
authors.initials: M
authors.lastname: Aziz
authors.forename: Ibtissam R
authors.initials: IR
authors.lastname: Hussein
mesh_terms: D000293:Adolescent; D000328:Adult; D017398:Alternative Splicing; D002648:Child; D002675:Child, Preschool; D004534:Egypt; D005260:Female; D005914:Globins / Q000235:genetics*; D006720:Homozygote; D006801:Humans; D007438:Introns; D008297:Male; D009154:Mutation; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D012322:RNA Precursors / Q000235:genetics*; D012326:RNA Splicing / Q000187:drug effects; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017086:beta-Thalassemia / Q000235:genetics*
chemical_list: D016376:Oligonucleotides, Antisense; D012322:RNA Precursors; D005914:Globins