An orthotopic bladder tumor model and the evaluation of intravesical saRNA treatment

J Vis Exp. 2012 Jul 28:(65):4207. doi: 10.3791/4207.

Abstract

We present a novel method for treating bladder cancer with intravesically delivered small activating RNA (saRNA) in an orthotopic xenograft mouse bladder tumor model. The mouse model is established by urethral catheterization under inhaled general anesthetic. Chemical burn is then introduced to the bladder mucosa using intravesical silver nitrate solution to disrupt the bladder glycosaminoglycan layer and allows cells to attach. Following several washes with sterile water, human bladder cancer KU-7-luc2-GFP cells are instilled through the catheter into the bladder to dwell for 2 hours. Subsequent growth of bladder tumors is confirmed and monitored by in vivo bladder ultrasound and bioluminescent imaging. The tumors are then treated intravesically with saRNA formulated in lipid nanoparticles (LNPs). Tumor growth is monitored with ultrasound and bioluminescence. All steps of this procedure are demonstrated in the accompanying video.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Administration, Intravesical
  • Animals
  • Cell Line, Tumor
  • Female
  • Humans
  • Luminescent Measurements / methods
  • Mice
  • Mice, Nude
  • Nanoparticles
  • RNA / administration & dosage*
  • RNA / genetics
  • Ultrasonography
  • Urinary Bladder Neoplasms / diagnostic imaging
  • Urinary Bladder Neoplasms / genetics
  • Urinary Bladder Neoplasms / pathology*
  • Urinary Bladder Neoplasms / therapy*
  • Xenograft Model Antitumor Assays / methods*

Substances

  • RNA