Fig. 5. Validation of anlotinib response in OMM organoid–derived subcutaneous xenografts in vivo.

(A) Representative images demonstrating the formation of xenografted tumors derived from OMM organoid lines. (B) A summary table of the xenografted tumors for organoid lines (details in table S3). (C) H&E and IHC staining of SOX10, S100, and Ki67 markers in xenografted tumors derived from OMM organoid lines. (D) Dose-response curves for anlotinib treatment in OMM2_OS1-P13 and OMM3_OLN-P17 organoids, using two forms of anlotinib: one for in vitro drug screening (anlotinib, research use only) and the other for in vivo xenografts (FuKeWei, anlotinib product). (E) Overview of the treatment schedule for OMM organoid–derived xenografts. (F) In vivo activity of anlotinib (2 mg/kg) in OMM2_OS1 organoid-derived xenografts, showing the tumor volumes at different time points. Mice were treated with the drug or vehicle daily for 28 days. *P < 0.05, Mann-Whitney test, two-tailed. (G) Bar plots illustrating the tumor weights of xenografts in the treated and control groups. (H) Kaplan-Meier curves for anlotinib- or vehicle-treated mice bearing OMM3_OLN xenografts. ns, not significant. (I) Bar plots presenting the tumor weights of OMM3_OLN xenografts in the anlotinib or vehicle control group. (J) H&E staining results displaying tumor-associated vessels and necrotic areas in OMM organoid–derived xenografts in the anlotinib/vehicle groups. CTRL, control. (K) Quantification of tumor-associated CD31-positive vessels to assess microvascular changes in OMM organoid–derived xenografts in response to anlotinib. OMM2_OS1 (left) and OMM3_OLN (right) (n = 5 animals per group, n = 10 high-power-field tumor areas per animal for statistical scoring). For (F) to (I), n = 5 in the anlotinib group; n = 6 in the vehicle group. Data in (F), (G), (I), and (K) are presented as the means ± SEM, and a two-tailed unpaired Student’s t test was used for the P values. *P < 0.05 and **P < 0.01.