Figure 8.

Intravenous administration of MECOM-depleted ADV/protein complex in orthotopic xenograft model

(A–C) A diagram (A) for intravenous administration of ADV/protein complex with MECOM depletion in H520 orthotopic xenograft (NOD-SCID mice). MECOM-depleted ADV was incubated with CFS and HF proteins to form ADV/protein complex for 2 h. The complex was injected into the mice via tail vein for 2 rounds with a 14-day interval. The ADV amount was 0.7 × 10¹⁰ VP, and the protein concentration was 1.0 × 10⁻⁷ pmol/VP. The bioluminescent imaging (BI) was taken on days 0, 14, and 28 after ADV delivery to demonstrate the tumor growth. Bioluminescent images (B) and growth curves (C) of orthotopic tumors treated with MECOM-depleted ADV/protein complex (MECOM-KO) via tail-vein injection. n = 4. ∗p ＜ 0.05, unpaired 2-tailed t test.

(D) Immunohistochemistry staining demonstrates MECOM, ABCG2, and CD44 expression in the H520 orthotopic xenografts treated with ADV/protein complex including control and MECOM depletion groups. Scale bar, 100 μm.

(E) Western blots show the indicated proteins in the H520 orthotopic xenografts treated with ADV/protein complex, including control and MECOM depletion groups. β-Actin was used as loading control. The numerical value under the band shows densitometric analyses of the indicated protein expression, compared to the corresponding control, which was normalized as “1.0.”

(F) On-target and off-target mutations detected by WES sequencing in tumor tissues of orthotopic models with MECOM depletion. The frequencies and locations of various on-target and off-target mutations were respectively presented in bar plots with different colors.

(G) The charts show the numbers of SNVs and indels detected in the tumors derived from H520 orthotopic xenograft with ADV/protein treatment.