title: CD4 aptamer-RORγt shRNA chimera inhibits IL-17 synthesis by human CD4(+) T cells.
abstract: Cell type specific delivery of RNAi to T cells has remained to be a challenge. Here we describe an aptamer mediated delivery of shRNA to CD4(+) T cells targeting RORγt to suppress Th17 cells. A cDNA encoding CD4 aptamer and RORγt shRNA was constructed and the chimeric CD4 aptamer-RORγt shRNA (CD4-AshR-RORγt) was generated using in vitro T7 RNA transcription. 2'-F-dCTP and 2'-F-dUTP were incorporated into CD4-AshR-RORγt for RNase resistance. CD4-AshR-RORγt was specifically uptaken by CD4(+) Karpas 299 cells and primary human CD4(+) T cells. The RORγt shRNA moiety of CD4-AshR-RORγt chimera was cleaved and released by Dicer. Furthermore, CD4-AshR-RORγt suppressed RORγt gene expression in Karpas 299 cells and CD4(+) T cells and consequently inhibited Th17 cell differentiation and IL-17 production. These results demonstrate that aptamer-facilitated cell specific delivery of shRNA represents a novel approach for efficient RNAi delivery and is potentially to be developed for therapeutics targeting specific T cells subtypes.
authors.affiliation: Division of Arthritis and Rheumatic Diseases, Oregon Health & Science University and Portland VA Medical Center, Portland, OR 97239, United States.
authors.forename: Pingfang
authors.initials: P
authors.lastname: Song
authors.affiliation: Division of Arthritis and Rheumatic Diseases, Oregon Health & Science University and Portland VA Medical Center, Portland, OR 97239, United States.
authors.forename: Yuan K
authors.initials: YK
authors.lastname: Chou
authors.affiliation: Division of Arthritis and Rheumatic Diseases, Oregon Health & Science University and Portland VA Medical Center, Portland, OR 97239, United States.
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Neuroimmunology Research, Portland VA Medical Center, Portland, OR 97239, United States.
authors.forename: Roberto
authors.initials: R
authors.lastname: Meza-Romero
authors.affiliation: Division of Arthritis and Rheumatic Diseases, Oregon Health & Science University and Portland VA Medical Center, Portland, OR 97239, United States; Department of Pediatrics, Oakland University William Beaumont Hospital, Royal Oak, MI 48073, United States.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Yomogida
authors.affiliation: Neuroimmunology Research, Portland VA Medical Center, Portland, OR 97239, United States.
authors.forename: Gil
authors.initials: G
authors.lastname: Benedek
authors.affiliation: Division of Arthritis and Rheumatic Diseases, Oregon Health & Science University and Portland VA Medical Center, Portland, OR 97239, United States. Electronic address: chuc@ohsu.edu.
authors.forename: Cong-Qiu
authors.initials: CQ
authors.lastname: Chu
mesh_terms: D015704:CD4 Antigens / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D006801:Humans; D020381:Interleukin-17 / Q000096:biosynthesis*; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3 / Q000235:genetics* / Q000276:immunology*; D034741:RNA, Small Interfering / Q000235:genetics*; D058504:Th17 Cells / Q000276:immunology*; D014162:Transfection / Q000379:methods
chemical_list: D015704:CD4 Antigens; D020381:Interleukin-17; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3; D034741:RNA, Small Interfering; C539900:RORC protein, human
keywords: Aptamer–shRNA; RORγt; Th17