CUT&Tag-seq Analysis of HCC-PDX Samples Treated with SP2509

Nude mice harboring HCC-PDX were subjected to treatment with either vehicle or SP2509, followed by analysis of tumor samples using CUT&Tag-seq assays. Tumor sample fragments (20-30 mm3) derived from an HCC patient into male nude mice (6 weeks old). After one month, twelve mice with tumors reaching an average size of approximately 163 mm3 were selected and randomly divided into 2 groups: six mice in the control group received intraperitoneal injection of 10μl/g vehicle (10% DMSO, 90% corn oil), and six mice in the test group received SP2509 treatment (i.p., 10μl/g, 25 mg/kg BW). Subsequently, tumor xenograft volumes and mice body weights were measured every three days for three weeks. The formula for calculating tumor volume was V = 0.5 × a × b2, where a and b represent the long and short diameters of the tumor, respectively. On the 21st day, mice were euthanized. The dissected tumor samples from PDX models were then subjected to the following real-time PCR assays and CUT&Tag-seq assays. This experimental protocol was approved by the Institutional Animal Care and Use Committee (IACUC) of Shanghai GeneChem Organism.