Selective gene silencing in activated leukocytes by targeting siRNAs to the integrin lymphocyte function-associated antigen-1

Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):4095-100. doi: 10.1073/pnas.0608491104. Epub 2007 Feb 28.

Abstract

Silencing gene expression by RNAi is a powerful method for exploring gene function and validating drug targets and potentially for therapy. Lymphocytes and other primary blood cells are resistant to lipid-based transfection in vitro and are difficult to target in vivo. We show here that antibody-protamine fusion proteins targeting the human integrin lymphocyte function-associated antigen-1 (LFA-1) efficiently deliver siRNAs and specifically induce silencing in primary lymphocytes, monocytes, and dendritic cells. Moreover, a fusion protein constructed from an antibody that preferentially recognizes activation-dependent conformational changes in LFA-1 selectively targets activated leukocytes and can be used to suppress gene expression and cell proliferation only in activated lymphocytes. The siRNA-fusion protein complexes do not cause lymphocyte activation or induce IFN responses. K562 cells expressing latent WT or constitutively activated LFA-1 engrafted in the lungs of SCID mice are selectively targeted by intravenously injected fusion protein-siRNA complexes, demonstrating the potential in vivo applicability of LFA-1-directed siRNA delivery.

Publication types

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

MeSH terms

  • Animals
  • Antigens, Nuclear / biosynthesis
  • Cell Adhesion
  • Cell Proliferation
  • DNA-Binding Proteins / biosynthesis
  • Dose-Response Relationship, Drug
  • Gene Silencing*
  • Humans
  • K562 Cells
  • Ku Autoantigen
  • Lymphocyte Function-Associated Antigen-1 / metabolism
  • Lymphocyte Function-Associated Antigen-1 / physiology*
  • Mice
  • Mice, SCID
  • RNA, Small Interfering / metabolism
  • Receptors, CCR5 / metabolism
  • Recombinant Fusion Proteins / chemistry
  • T-Lymphocytes / metabolism

Substances

  • Antigens, Nuclear
  • DNA-Binding Proteins
  • Lymphocyte Function-Associated Antigen-1
  • RNA, Small Interfering
  • Receptors, CCR5
  • Recombinant Fusion Proteins
  • Xrcc6 protein, human
  • Xrcc6 protein, mouse
  • Ku Autoantigen