title: Defective antiviral responses of induced pluripotent stem cells to baculoviral vector transduction.
abstract: Genetic engineering of induced pluripotent stem cells (iPSCs) is important for their clinical applications, and baculovirus (BV) holds promise as a gene delivery vector. To explore the feasibility of using BV for iPSCs transduction, in this study we first examined how iPSCs responded to BV. We determined that BV transduced iPSCs efficiently, without inducing appreciable negative effects on cell proliferation, apoptosis, pluripotency, and differentiation. BV transduction slightly perturbed the transcription of 12 genes involved in the Toll-like receptor (TLR) signaling pathway, but at the protein level BV elicited no well-known cytokines (e.g., interleukin-6 [IL-6], tumor necrosis factor alpha [TNF-α], and beta interferon [IFN-β]) except for IP-10. Molecular analyses revealed that iPSCs expressed no TLR1, -6, -8, or -9 and expressed merely low levels of TLR2, -3, and -4. In spite of evident expression of such RNA/DNA sensors as RIG-I and AIM2, iPSCs barely expressed MDA5 and DAI (DNA-dependent activator of IFN regulatory factor [IRF]). Importantly, BV transduction of iPSCs stimulated none of the aforementioned sensors or their downstream signaling mediators (IRF3 and NF-κB). These data together confirmed that iPSCs responded poorly to BV due to the impaired sensing and signaling system, thereby justifying the transduction of iPSCs with the baculoviral vector.
authors.affiliation: Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Guan-Yu
authors.initials: GY
authors.lastname: Chen
authors.forename: Shiaw-Min
authors.initials: SM
authors.lastname: Hwang
authors.forename: Hung-Ju
authors.initials: HJ
authors.lastname: Su
authors.forename: Chien-Yi
authors.initials: CY
authors.lastname: Kuo
authors.forename: Wen-Yi
authors.initials: WY
authors.lastname: Luo
authors.forename: Kai-Wei
authors.initials: KW
authors.lastname: Lo
authors.forename: Cheng-Chieh
authors.initials: CC
authors.lastname: Huang
authors.forename: Chiu-Ling
authors.initials: CL
authors.lastname: Chen
authors.forename: Sheng-Han
authors.initials: SH
authors.lastname: Yu
authors.forename: Yu-Chen
authors.initials: YC
authors.lastname: Hu
mesh_terms: D000818:Animals; D017209:Apoptosis; D016367:Baculoviridae / Q000276:immunology*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D016207:Cytokines / Q000276:immunology; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases / Q000276:immunology; D004268:DNA-Binding Proteins; D005822:Genetic Vectors / Q000276:immunology*; D006023:Glycoproteins / Q000276:immunology; D057026:Induced Pluripotent Stem Cells / Q000276:immunology* / Q000473:pathology; D050838:Interferon Regulatory Factor-3 / Q000276:immunology; D000072640:Interferon-Induced Helicase, IFIH1; D051379:Mice; D016328:NF-kappa B / Q000276:immunology; D009687:Nuclear Proteins / Q000276:immunology; D016601:RNA-Binding Proteins; D051193:Toll-Like Receptors / Q000276:immunology; D014161:Transduction, Genetic
chemical_list: C540422:Aim2 protein, mouse; D016207:Cytokines; D004268:DNA-Binding Proteins; D006023:Glycoproteins; D050838:Interferon Regulatory Factor-3; C494233:Irf3 protein, mouse; D016328:NF-kappa B; D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; D051193:Toll-Like Receptors; C402632:Zbp1 protein, mouse; C503356:Ddx58 protein, mouse; C510592:Ifih1 protein, mouse; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases; D000072640:Interferon-Induced Helicase, IFIH1