title: Enhancement of β-Globin Gene Expression in Thalassemic IVS2-654 Induced Pluripotent Stem Cell-Derived Erythroid Cells by Modified U7 snRNA.
abstract: The therapeutic use of patient-specific induced pluripotent stem cells (iPSCs) is emerging as a potential treatment of β-thalassemia. Ideally, patient-specific iPSCs would be genetically corrected by various approaches to treat β-thalassemia including lentiviral gene transfer, lentivirus-delivered shRNA, and gene editing. These corrected iPSCs would be subsequently differentiated into hematopoietic stem cells and transplanted back into the same patient. In this article, we present a proof of principle study for disease modeling and screening using iPSCs to test the potential use of the modified U7 small nuclear (sn) RNA to correct a splice defect in IVS2-654 β-thalassemia. In this case, the aberration results from a mutation in the human β-globin intron 2 causing an aberrant splicing of β-globin pre-mRNA and preventing synthesis of functional β-globin protein. The iPSCs (derived from mesenchymal stromal cells from a patient with IVS2-654 β-thalassemia/hemoglobin (Hb) E) were transduced with a lentivirus carrying a modified U7 snRNA targeting an IVS2-654 β-globin pre-mRNA in order to restore the correct splicing. Erythroblasts differentiated from the transduced iPSCs expressed high level of correctly spliced β-globin mRNA suggesting that the modified U7 snRNA was expressed and mediated splicing correction of IVS2-654 β-globin pre-mRNA in these cells. Moreover, a less active apoptosis cascade process was observed in the corrected cells at transcription level. This study demonstrated the potential use of a genetically modified U7 snRNA with patient-specific iPSCs for the partial restoration of the aberrant splicing process of β-thalassemia. Stem Cells Translational Medicine 2017;6:1059-1069.
authors.affiliation: Departments of Anatomy.|Stem Cell Research Group.
authors.forename: Phetcharat
authors.initials: P
authors.lastname: Phanthong
authors.affiliation: Biotechnology.|Excellent Center for Drug Discovery, Faculty of Science, Mahidol University, Bangkok, Thailand.
authors.forename: Suparerk
authors.initials: S
authors.lastname: Borwornpinyo
authors.affiliation: Stem Cell Research Group.
authors.forename: Narisorn
authors.initials: N
authors.lastname: Kitiyanant
authors.affiliation: Thalassemia Research Center.
authors.forename: Natee
authors.initials: N
authors.lastname: Jearawiriyapaisarn
authors.affiliation: Departments of Obstetrics and Gynecology.
authors.forename: Lalana
authors.initials: L
authors.lastname: Nuntakarn
authors.affiliation: Anatomy Department, Faculty of Science, Prince of Songkla University, Songkhla, Thailand.
authors.forename: Jirawat
authors.initials: J
authors.lastname: Saetan
authors.affiliation: Thalassemia Research Center.
authors.forename: Tiwaporn
authors.initials: T
authors.lastname: Nualkaew
authors.affiliation: Department of Biochemistry, Faculty of Pharmacy, Mahidol University, Bangkok, Thailand.
authors.forename: Khanit
authors.initials: K
authors.lastname: Sa-Ngiamsuntorn
authors.affiliation: Pediatrics, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand.
authors.forename: Usanarat
authors.initials: U
authors.lastname: Anurathapan
authors.affiliation: Biotalentum Ltd, Godollo, Hungary.|Molecular Animal Biotechnology Laboratory, Szent Istvan University, Godollo, Hungary.
authors.forename: Andras
authors.initials: A
authors.lastname: Dinnyes
authors.affiliation: Departments of Anatomy.|Stem Cell Research Group.|Reproductive Biology Research Group, Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom, Thailand.
authors.forename: Yindee
authors.initials: Y
authors.lastname: Kitiyanant
authors.affiliation: Pediatrics, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand.
authors.forename: Suradej
authors.initials: S
authors.lastname: Hongeng
mesh_terms: D000818:Animals; D041905:Erythroid Cells / Q000166:cytology* / Q000378:metabolism*; D015870:Gene Expression / Q000235:genetics*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D012326:RNA Splicing / Q000235:genetics / Q000502:physiology; D012342:RNA, Small Nuclear / Q000235:genetics*; D059467:Transcriptome / Q000235:genetics; D055544:beta-Globins / Q000235:genetics*; D017086:beta-Thalassemia / Q000235:genetics / Q000378:metabolism
chemical_list: D012342:RNA, Small Nuclear; C405487:U7 small nuclear RNA; D055544:beta-Globins
keywords: Induced pluripotent stem cell; Induced pluripotent stem cell-derived erythroblast; U7 snRNA; β-Thalassemia