title: Amphoteric liposomes enable systemic antigen-presenting cell-directed delivery of CD40 antisense and are therapeutically effective in experimental arthritis.
abstract: OBJECTIVE
  Mediation of RNA interference by oligonucleotides constitutes a powerful approach for the silencing of genes involved in the pathogenesis of inflammatory disease, but in vivo application of this technique requires effective delivery to immune cells and/or sites of inflammation. The aim of the present study was to develop a new carrier system to mediate systemic administration of oligonucleotides to rheumatoid arthritis (RA) joints, and to develop an antisense oligonucleotide (ASO)-based approach to interfere with CD40-CD154 interactions in an experimental model of RA.
  
  
  METHODS
  A novel liposomal carrier with amphoteric properties, termed Nov038, was developed and assessed for its ability to systemically deliver an ASO directed against CD40 (CD40-ASO). Male DBA/1 mice with collagen-induced arthritis were treated with Nov038-encapsulated CD40-ASO, and the effects of treatment on various parameters of disease activity, including clinical score, paw swelling, lymph node responses, and inflammatory cytokine production in the joints, were assessed.
  
  
  RESULTS
  Nov038 was well tolerated, devoid of immune-stimulatory effects, and efficacious in mediating systemic oligonucleotide delivery to sites of inflammation. In mice with collagen-induced arthritis, Nov038 enabled the therapeutic administration of CD40-ASO and improved established disease, while unassisted CD40-ASO was ineffective, and anti-tumor necrosis factor alpha (anti-TNFalpha) treatment was less effective in this model. Nov038/CD40-ASO efficacy was attributed to its tropism for monocyte/macrophages and myeloid dendritic cells (DCs), resulting in rapid down-regulation of CD40, inhibition of DC antigen presentation, and reduction in collagen-specific T cell responses, as well as decreased levels of TNFalpha, interleukin-6 (IL-6), and IL-17 in arthritic joints.
  
  
  CONCLUSIONS
  Amphoteric liposomes represent a novel carrier concept for systemic and antigen-presenting cell-targeted oligonucleotide delivery with clinical applicability and numerous potential applications, including target validation in vivo and inflammatory disease therapeutics. Moreover, Nov038/CD40-ASO constitutes a potent alternative to monoclonal antibody-based approaches for interfering with CD40-CD40L interactions.
authors.affiliation: Biomedical Research Foundation, Academy of Athens, Athens, Greece. vandreakos@bioacademy.gr
authors.forename: Evangelos
authors.initials: E
authors.lastname: Andreakos
authors.forename: Una
authors.initials: U
authors.lastname: Rauchhaus
authors.forename: Athanassios
authors.initials: A
authors.lastname: Stavropoulos
authors.forename: Gerold
authors.initials: G
authors.lastname: Endert
authors.forename: Volkmar
authors.initials: V
authors.lastname: Wendisch
authors.forename: Amina S
authors.initials: AS
authors.lastname: Benahmed
authors.forename: Stavros
authors.initials: S
authors.lastname: Giaglis
authors.forename: James
authors.initials: J
authors.lastname: Karras
authors.forename: Sam
authors.initials: S
authors.lastname: Lee
authors.forename: Hans
authors.initials: H
authors.lastname: Gaus
authors.forename: C Frank
authors.initials: CF
authors.lastname: Bennett
authors.forename: Richard O
authors.initials: RO
authors.lastname: Williams
authors.forename: Paschalis
authors.initials: P
authors.lastname: Sideras
authors.forename: Steffen
authors.initials: S
authors.lastname: Panzner