title: A genetic strategy to treat sickle cell anemia by coregulating globin transgene expression and RNA interference.
abstract: The application of RNA interference (RNAi) to stem cell-based therapies will require highly specific and lineage-restricted gene silencing. Here we show the feasibility and therapeutic potential of coregulating transgene expression and RNAi in hematopoietic stem cells. We encoded promoterless small-hairpin RNA (shRNA) within the intron of a recombinant gamma-globin gene. Expression of both gamma-globin and the lariat-embedded small interfering RNA (siRNA) was induced upon erythroid differentiation, specifically downregulating the targeted gene in tissue- and differentiation stage-specific fashion. The position of the shRNA within the intron was critical to concurrently achieve high-level transgene expression, effective siRNA generation and minimal interferon induction. Lentiviral transduction of CD34(+) cells from patients with sickle cell anemia led to erythroid-specific expression of the gamma-globin transgene and concomitant reduction of endogenous beta(S) transcripts, thus providing proof of principle for therapeutic strategies that require synergistic gene addition and gene silencing in stem cell progeny.
authors.affiliation: Laboratory of Gene Transfer and Gene Expression, Sloan-Kettering Institute, New York, New York 10021, USA.
authors.forename: Selda
authors.initials: S
authors.lastname: Samakoglu
authors.forename: Leszek
authors.initials: L
authors.lastname: Lisowski
authors.forename: Tulin
authors.initials: T
authors.lastname: Budak-Alpdogan
authors.forename: Yelena
authors.initials: Y
authors.lastname: Usachenko
authors.forename: Santina
authors.initials: S
authors.lastname: Acuto
authors.forename: Rosalba
authors.initials: R
authors.lastname: Di Marzo
authors.forename: Aurelio
authors.initials: A
authors.lastname: Maggio
authors.forename: Ping
authors.initials: P
authors.lastname: Zhu
authors.forename: John F
authors.initials: JF
authors.lastname: Tisdale
authors.forename: Isabelle
authors.initials: I
authors.lastname: Rivière
authors.forename: Michel
authors.initials: M
authors.lastname: Sadelain
mesh_terms: D000755:Anemia, Sickle Cell / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology / Q000628:therapy; D002478:Cells, Cultured; D005786:Gene Expression Regulation / Q000235:genetics; D020868:Gene Silencing; D018390:Gene Targeting / Q000379:methods; D015316:Genetic Therapy / Q000379:methods*; D005914:Globins / Q000235:genetics* / Q000378:metabolism* / Q000627:therapeutic use; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D034622:RNA Interference; D016896:Treatment Outcome
chemical_list: D005914:Globins