In Vivo Tumor Growth Study in PDX Models

The in vivo tumor growth study was performed in accordance with the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health and approved by MD Anderson’s Institutional Animal Care and Use Committee. The PDX models were generated as described previously (17 (link)) and subjected to qPCR as described to determine the HPV type (18 (link),19 (link)). Briefly, tumor tissue was cut into small fragments (5–6 mm) and implanted subcutaneously into the flanks of nude mice. The skin incisions were closed with skin clips that were removed after 10 to 15 days. Once tumors reached an average of 150 to 300 mm³, the mice were randomized into either vehicle group or Alisertib group. Alisertib (Takeda Pharmaceuticals, Lexington, MA; 10 mg/kg in 10% β-cyclodextrin) was administered via oral gavage once daily on a weekly schedule of 6 days on and 1 day off. Tumors were monitored daily, and tumor volume (length × width² × 0.5) was evaluated twice per week with digital calipers. Mice were euthanized when tumors reached 2000 mm³. The endpoint for the survival studies was the tumor burden. Survival was measured using the Kaplan-Meier method (20 (link)). Survival curve analysis was performed with GraphPad Prism software version 9 (RRID:SCR_002798).