title: Synergistic inhibition of PARP-1 and NF-κB signaling downregulates immune response against recombinant AAV2 vectors during hepatic gene therapy.
abstract: Host immune response remains a key obstacle to widespread application of adeno-associated virus (AAV) based gene therapy. Thus, targeted inhibition of the signaling pathways that trigger such immune responses will be beneficial. Previous studies have reported that DNA damage response proteins such as poly(ADP-ribose) polymerase-1 (PARP-1) negatively affect the integration of AAV in the host genome. However, the role of PARP-1 in regulating AAV transduction and the immune response against these vectors has not been elucidated. In this study, we demonstrate that repression of PARP-1 improves the transduction of single-stranded AAV vectors both in vitro (∼174%) and in vivo (two- to 3.4-fold). Inhibition of PARP-1, also significantly downregulated the expression of several proinflammatory and cytokine markers such as TLRs, ILs, NF-κB subunit proteins associated with the host innate response against self-complementary AAV2 vectors. The suppression of the inflammatory response targeted against these vectors was more effective upon combined inhibition of PARP-1 and NF-κB signaling. This strategy also effectively attenuated the AAV capsid-specific cytotoxic T-cell response, with minimal effect on vector transduction, as demonstrated in normal C57BL/6 and hemophilia B mice. These data suggest that targeting specific host cellular proteins could be useful to attenuate the immune barriers to AAV-mediated gene therapy.
authors.affiliation: Centre for Stem Cell Research, Christian Medical College, Vellore, Tamil Nadu, India.
authors.forename: Sangeetha
authors.initials: S
authors.lastname: Hareendran
authors.affiliation: Department of General Pathology, Christian Medical College, Vellore, Tamil Nadu, India.
authors.forename: Banumathi
authors.initials: B
authors.lastname: Ramakrishna
authors.affiliation: Centre for Stem Cell Research, Christian Medical College, Vellore, Tamil Nadu, India.|Department of Hematology, Christian Medical College, Vellore, Tamil Nadu, India.|Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur, Uttar Pradesh, India.
authors.forename: Giridhara R
authors.initials: GR
authors.lastname: Jayandharan
mesh_terms: D000818:Animals; D015153:Blotting, Western; D000229:Dependovirus / Q000276:immunology*; D004195:Disease Models, Animal; D015536:Down-Regulation; D004357:Drug Synergism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D015316:Genetic Therapy / Q000009:adverse effects* / Q000379:methods; D005822:Genetic Vectors / Q000009:adverse effects* / Q000276:immunology; D006367:HeLa Cells; D002836:Hemophilia B / Q000628:therapy; D006801:Humans; D008099:Liver / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000276:immunology; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases / Q000276:immunology*; D060888:Real-Time Polymerase Chain Reaction; D014161:Transduction, Genetic; D014162:Transfection
chemical_list: D004791:Enzyme Inhibitors; D016328:NF-kappa B; C499320:PARP1 protein, human; C499321:Parp1 protein, mouse; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases
keywords: AAV; Gene therapy; Hemophilia; Immune response; PARP-1