Abstract P5-06-01: Proteomic analysis of conserved kinases between PDX tumors and corresponding PDX-derived cell lines
Authors: Han A , Kim B-J , Chan DW , Matsunuma R , Singh P , Ellis MJ 
Published: February 2018

(Background) Patient-derived xenograft (PDX)s are valuable models for precision oncology as they are thought to recapitulate the biological and genomic characteristics of the human tumors they were derived from. Even though this model is widely used, it is still difficult to manipulate genes in this system and it is not as convenient as in vitro cell culture for drug sensitivity screening. On the other hand in vitro cell cultures have a highly artificial microenvironment and also have undergone selection which might generate misleading data. To address these issues cell lines from PDX tumors with different intrinsic subtypes were characterized by kinobead precipitation/mass spectrometry analysis (KiP/MS) to profile each PDX line in vivo and ex vivo for therapeutic targets.

(Materials and Methods)Washington University Human in Mice (WHIM) tumors and Hutsman Cancer Institute (HCI) tumors were transplanted into mammary fat pads of female severe combined immunodeficiency/beige (SCID/beige) mice. PDXs tumors were harvested when they reached 1˜1.5cm. PDX tumor-derived ex vivo cells/organoids were routinely cultured with Rock inhibitor support. Estradiol was applied only for the tumors originating from E2 supplementation in vivo. Cells and tumors were harvested and lysed by sonication. Kinases in soluble lysates are enriched with drug-bound beads (kinobead) and digested with trypsin. Digested peptides are analyzed by mass spectrometry.

(Results) 5 of WHIM tumors and 4 of HCI tumors were successfully dissociated and cultured ex vivo for further analysis and experiments. Hierarchical clustering of tumors and corresponding cells showed ex vivo cultured cells cluster together with their original PDX tumors. Tumors/ex vivo cultured cells clustered by intrinsic subtype and enrichment analysis identified specific kinases for each PDX tumor/cell line. The WHIM 4 tumor-cell pair showed high level of PIM kinase and EGFR and other PDX tumors such as WHIM18, WHIM 20 (Luminal subtypes), WHIM 35(HER2 enriched subtype) also showed model unique intrinsic kinases, such as JAK2 for WHIM18, EPHB4 for WHIM20.

(Conclusion) PDX tumor-derived ex vivo lines could be routinely cultured with Rock inhibitor support. Each PDX tumor and cell line pair were cluster together in hierarchical clustering and categorized into the same intrinsic subtype based on kinome profiling, suggesting these cells maintain their tumor specific intrinsic kinase signaling. A subset of kinases exhibit activity/expression that is conserved after ex vivo culture, we hypothesize these are intrinsic kinases might be promising target for treatment, because they are tumor intrinsic, i.e. their high expression is maintained despite the strong contrast in the microenvironment of in vitro versus in vitro growth. Ongoing studies with drugs and knock down reagents are examining whether the in vivo/ex vivo comparative KIP analysis indeed identifies therapeutic targets, which will be presented at the meeting more in detail.

Citation Format: Han A, Kim B-J, Chan DW, Matsunuma R, Singh P, Ellis MJ. Proteomic analysis of conserved kinases between PDX tumors and corresponding PDX-derived cell lines [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P5-06-01.