Figure 2.

Sonodelivery of Gluc Fusion Proteins In Vivo

(A) Schematic of sonodelivery for expressing Gaussia luciferase (Gluc) proteins in mouse muscle. Nan Nanoplex formation with the rNLSd polumer was performed as described in Takeda et al.,¹¹ complexed with plasmid DNA encoding Gluc. NN This nanoplex is delivered in the presence of microbubbles (MB) as described in Materials and Methods. An ultrasound stimulus (US) is applied to disrupt the MB and the nanoplex of polymer:pGluc mediates skeletal muscle cell transfection. The proteins secreted contain a C-terminal peptide tag that either targets the IL-6Rα (pepL) or is untargeted (non-specific peptide control). (B) Ex vivo Gluc imaging post-gene delivery. Bioluminescence imaging is shown using coelenterazine substrate on organs isolated from animals receiving control (Gluc.ns) or ligand targeted Gluc (Gluc.pepL). Color bar, p/sec/cm²/sr. Signals are present in the tumor:bone region only when targeted Gluc.pepL is delivered to muscle. Right plot, average Gluc signals from tumor:bone ex vivo pooled from day 10 and 14 post-delivery (∗p < 0.013 for comparing Gluc.pepL relative to Gluc.ns accumulation in tumor tissue (accumulation in normal organs not significantly different between Gluc.ns and Gluc.pepL). Normal organs included an imaged pool of either liver, lung and heart (Li, Lu, H) or kidney, small intestine, spleen, and pancreas (Kid, Int, Spl, Panc). Mice were bearing TC2R tumors intratibially. Data is expressed as mean +- the standard error of the mean.