Figure 9.

Therapeutic effect of K27-D30 encapsulating LNPs on tumor growth in an HTVI-induced mouse model of hepatocellular carcinoma. (A) Experimental scheme. Mice received HTVI on day 0, began receiving treatment week 5, and were evaluated for tumor growth at an endpoint of 7 or 8 weeks for the non-luciferase and luciferase expressing subsets, respectively. (B) Tumor counts and liver weights (as a percentage of total body weight) at endpoint for: healthy (received LR only during HTVI at day 0), PBS (received PBS injections starting week 5), free protein (received 4.5 mg/kg of K27-D30 2x/week starting week 5), and LNP (received 4.5 mg/kg of LNP-encapsulated K27-D30 2x/week starting week 5) groups. n = 5 for LR and n ≧ 9 for others; *: p <0.05, **: p <0.01. Group receiving LNPs shows significantly reduced tumor counts as compared to PBS control. (C) Luminescence (collected using IVIS) of whole body and extracted livers of mice that received an additional luciferase reporter gene during HTVI. Both free protein and LNP groups show reduction in luciferase signal over PBS control, indicating reduced luciferase production in these tumors. n ≧ 4; *: p <0.05, ***: p <0.001, and ****: p <0.0001. (D) Representative images of H&E stained liver sections showing tumors in LR (healthy), PBS, free protein, and LNP groups. Fewer, smaller, and more dispersed tumors—visible here by change in color—were seen in the LNP group, when compared to free protein and PBS. Scale bar = 1000μm. (E) Serum levels of alpha-fetoprotein (AFP), which indicate the presence of primary liver cancer. At midpoint (week 5), all groups are statistically similar. At endpoint (week 7/8), PBS group is elevated compared to healthy control, and LNP group is significantly reduced, when compared to PBS control. n = 5 for LR and n ≧ 8 for others; *: p <0.05, **: p <0.01