Sequence- and target-independent angiogenesis suppression by siRNA via TLR3

Nature. 2008 Apr 3;452(7187):591-7. doi: 10.1038/nature06765. Epub 2008 Mar 26.

Abstract

Clinical trials of small interfering RNA (siRNA) targeting vascular endothelial growth factor-A (VEGFA) or its receptor VEGFR1 (also called FLT1), in patients with blinding choroidal neovascularization (CNV) from age-related macular degeneration, are premised on gene silencing by means of intracellular RNA interference (RNAi). We show instead that CNV inhibition is a siRNA-class effect: 21-nucleotide or longer siRNAs targeting non-mammalian genes, non-expressed genes, non-genomic sequences, pro- and anti-angiogenic genes, and RNAi-incompetent siRNAs all suppressed CNV in mice comparably to siRNAs targeting Vegfa or Vegfr1 without off-target RNAi or interferon-alpha/beta activation. Non-targeted (against non-mammalian genes) and targeted (against Vegfa or Vegfr1) siRNA suppressed CNV via cell-surface toll-like receptor 3 (TLR3), its adaptor TRIF, and induction of interferon-gamma and interleukin-12. Non-targeted siRNA suppressed dermal neovascularization in mice as effectively as Vegfa siRNA. siRNA-induced inhibition of neovascularization required a minimum length of 21 nucleotides, a bridging necessity in a modelled 2:1 TLR3-RNA complex. Choroidal endothelial cells from people expressing the TLR3 coding variant 412FF were refractory to extracellular siRNA-induced cytotoxicity, facilitating individualized pharmacogenetic therapy. Multiple human endothelial cell types expressed surface TLR3, indicating that generic siRNAs might treat angiogenic disorders that affect 8% of the world's population, and that siRNAs might induce unanticipated vascular or immune effects.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cell Line
  • Endothelial Cells / metabolism
  • Genetic Therapy / methods*
  • Humans
  • Immunity, Innate / immunology*
  • Interferon-gamma / immunology
  • Interleukin-12 / immunology
  • Macular Degeneration / complications
  • Macular Degeneration / genetics
  • Macular Degeneration / therapy
  • Mice
  • Mice, Inbred C57BL
  • Neovascularization, Pathologic / genetics
  • Neovascularization, Pathologic / immunology*
  • Neovascularization, Pathologic / prevention & control*
  • Neovascularization, Pathologic / therapy
  • RNA, Small Interfering / chemistry
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / immunology*
  • RNA, Small Interfering / metabolism*
  • Toll-Like Receptor 3 / chemistry
  • Toll-Like Receptor 3 / genetics
  • Toll-Like Receptor 3 / metabolism*
  • Vascular Endothelial Growth Factor A / genetics

Substances

  • RNA, Small Interfering
  • Toll-Like Receptor 3
  • Vascular Endothelial Growth Factor A
  • Interleukin-12
  • Interferon-gamma