RNAseq data LUN 156 parental, adagrasib resistant and RMC-4998 resistant LUN156 tumors

LUN156 is a patient-derived xenograft (PDX) tumor model derived from 73-year-old male Chinese non-small cell lung cancer (NSCLC) patient with a tumor with a KRASG12C mutation. Overall design: PDX model development: To generate an animal tumor model, fresh tumor fragments were obtained from hospitals with informed consents from the patients in accordance with protocols approved by the Hospital Institutional Ethical Committee (IEC). The tumors were cut into 30 to 60 mm3 fragments after arrival at CRO animal facility and serially passaged after implanting subcutaneously into BALB/c nude mice. Briefly, five to six-weeks-old female Balb/c nude mice were purchased from Vital River Lab Animal Technology Co. Ltd., Beijing, China. Animals were held for a minimum of 7 days for acclimation prior to the beginning of study. Animals were housed in individually ventilated cages (IVC) under specific pathogen free (SPF) conditions with access to standard diet and water ad libitum. During the study period, the animals were monitored daily for any effects of tumor growth and treatments on normal behavior, such as mobility, food, and water consumption. Study-related parameters like body weight and tumor volume (tumor volume = ((width)2 × length)/2, where width and length are measured in mm) were measured twice weekly. Recovered tumor fragments of about 15-30 mm3 in volume were implanted subcutaneously into right flanks of Balb/c female nude mice and mice were randomized by blocks when average tumor sizes reached ~200 mm3 in volume into three treatment groups with three mice in each group. The animals were administered vehicle for RMC-4998 formulation as a control, RMC-4998 100 mg/kg, and adagrasib 100 mg/kg as a daily treatment through oral gavage for 28 days, and end of study tumor samples were collected. In an independent study, to derive a stable adagrasib or RMC-4998 resistant (adagrasib-R or RMC-4998-R) LUN156 PDX tumor model, animals were treated with 100 mg/kg of each inhibitor for 90 days to determine on-treatment tumor relapse. After treatment period, the mouse showing on-treatment tumor relapse was collected (Passage 0 (P0)). The 30-50 mm3 volume tumor fragments from P0 were further subcutaneously implanted into the right flanks of female Balb/c nude mice and allowed to grow. Once the tumor became palpable (70-100 mm3), respective KRASG12C inhibitor treatments were provided daily and observed for tumor growth until the tumor reached an average of 1,000 mm3 in volume (P1). The animal from each inhibitor resistant cohort showing the least doubling time for a tumor to grow was selected and passaged further to generate P2. The in vivo serial passage procedure with respective KRASG12C inhibitor was repeated until P5 to ensure resistant phenotype in LUN156 tumor model, and samples were frozen for efficacy and single dose pharmacodynamic use. During these passages, snap frozen tumor fragments were collected for resistance mechanism analyses