Ultrasound-based molecular imaging and specific gene delivery to mesenteric vasculature by endothelial adhesion molecule targeted microbubbles in a mouse model of Crohn's disease

J Control Release. 2013 Feb 10;165(3):216-25. doi: 10.1016/j.jconrel.2012.10.021. Epub 2012 Nov 8.

Abstract

Crohn's disease (CD) is a chronic inflammatory disorder of the gastrointestinal tract (GI) for which treatments with immunosuppressive drugs have significant side-effects. Consequently, there is a clinical need for site-specific and non-toxic delivery of therapeutic genes or drugs for CD and related disorders such as inflammatory bowel disease. The aim of this study was to validate a gene delivery platform based on ultrasound-activated lipid-shelled microbubbles (MBs) targeted to inflamed mesenteric endothelium in the CD-like TNFΔARE mouse model. MBs bearing luciferase plasmid were functionalized with antibodies to MAdCAM-1 (MB-M) or VCAM-1 (MB-V), biomarkers of gut endothelial cell inflammation and evaluated in an in vitro flow chamber assay with appropriate ligands to confirm targeting specificity. Following MB retro-orbital injection in TNFΔARE mice, the mean contrast intensity in the ileocecal region from accumulated MB-M and MB-V was 8.5-fold and 3.6-fold greater, respectively, compared to MB-C. Delivery of luciferase plasmid to the GI tract in TNFΔARE mice was achieved by insonating the endothelial cell-bound agents using a commercial sonoporator. Luciferase expression in the midgut was detected 48 h later by bioluminescence imaging and further confirmed by immunohistochemical staining. The liver, spleen, heart, and kidney had no detectable bioluminescence following insonation. Transfection of the microcirculation guided by a targeted, acoustically-activated platform such as an ultrasound contrast agent microbubble has the potential to be a minimally-invasive treatment strategy to ameliorate CD and other inflammatory conditions.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cell Adhesion Molecules / metabolism*
  • Crohn Disease / diagnostic imaging
  • Crohn Disease / metabolism*
  • DNA / administration & dosage*
  • Disease Models, Animal
  • Endothelium, Vascular / metabolism
  • Female
  • Gene Transfer Techniques*
  • Luciferases, Firefly / genetics
  • Luciferases, Firefly / metabolism
  • Mesentery / metabolism
  • Mice
  • Mice, Transgenic
  • Microbubbles*
  • Molecular Imaging
  • Plasmids
  • Tumor Necrosis Factor-alpha / genetics
  • Ultrasonography
  • Vascular Cell Adhesion Molecule-1 / metabolism*

Substances

  • Cell Adhesion Molecules
  • Madcam1 protein, mouse
  • Tumor Necrosis Factor-alpha
  • Vascular Cell Adhesion Molecule-1
  • DNA
  • Luciferases, Firefly