Abstract PD9-12: Integrative molecular profiling of breast cancer brain metastasis and patient-derived xenograft organoids from resected breast cancer brain metastases to interrogate and prioritize therapeutic personalized strategies
Authors: Morikawa A , Robinson DR , Soellner M , Wu Y-M , Lonigro R , Gilani R , Cheng X , Lachacz E , Thomas D , McMurray K , Smerage J , Henry NL , Heth J , Chinnaiyan A , Hayes DF , Merajver S 
Published: May 2019

Abstract
Background: Breast cancer brain metastasis (BM) is an area of unmet need in metastatic breast cancer patients. Novel therapeutic interventions to help prevent and treat BM are warranted. We conducted integrative molecular profiling of BM and matched primary tumors (PT) using next-generation DNA and RNA sequencing to examine the molecular landscape. In addition, we established patient-derived xenograft/organoid (PDX/PDO) to examine drug sensitivity according to the molecular and clinical features of the BM.
Methods: Archived, formalin fixed paraffin-embedded BM was collected retrospectively. BM were also collected prospectively at the time of clinically indicated surgical resection through the central nervous system tissue banking and the Michigan Oncology Sequencing Center (MI-ONCOSEQ) protocols. Matched archived PT tissues were collected when available. Integrative next-generation sequencing was conducted using the MI-ONCOSEQ platform. The prospectively collected BM were further used to establish PDXs/ PDOs. Successfully established PDXs/PDOs were used for ex vivo drug testing via MiDrugScreen, a novel drug sensitivity testing platform, where testing was performed in a dose-response format with drug selection prioritized by clinical scenario and molecular alterations if known a priori.
Results: 12 matched BM-PT pairs were analyzed: 6 triple negative, 5 HER2 positive, and 1 ER positive HER2 negative. All except one (11/12) had TP53 mutations. When present, TP53 mutations in BM were also found in PT (except for 1 unknwon case in PT due to low coverage). ER+HER2- was the only one without TP53 mutation but had hyper-mutation (APOBEC signature). Driver mutations and unique copy number alterations (CDKN2A loss in 1/12, mutations in PIK3CA in 1/12 and ESR1 in 1/12, CCNE1 amplification in 1/12) were noted in BMs. In 75% of cases, mutational burden was higher in BM vs. PT. 2 PDX/PDO were available for drug testing. PDO-BC9 was noted to have RB1 (splice acceptor) and LOH. As predicted by this alteration, PDO-BC9 was insensitive to CDK4/6 inhibitors (palbociclib, abemaciclib) tested on MiDrugScreen panel. PDX-BC4 was established from PIK3CA and ESR1 mutated BM from an ER+HER2- patient who had previously progressed on endocrine therapy with a CDK4/6 inhibitor. As predicted, the PDX-BC4 was resistant to CDK4/6 inhibitor but interestingly sensitive to PIK3CA, ERK, and MEK inhibitors.
Conclusions: TP53 mutation was highly prevalent and may be a biomarker for increased risk of BM. Further study is warranted to see if specific TP53 mutations are associated with a risk of BM development and can be used in risk stratification for BM specific intervention. Unique molecular alterations in BM compared to matched PT may have a therapeutic implication as a target or resistance biomarker. Conducting drug testing in addition to molecular profiling has the strong potential of being informative in tailoring or prioritizing therapeutic agents in the era of precision medicine. Additional BM PDXs/PDOs from breast and other solid tumors are being examined using this novel therapeutic tailoring approach with the combination of MIONCOSEQ and MiDrugScreen.
Citation Format: Morikawa A, Robinson DR, Soellner M, Wu Y-M, Lonigro R, Gilani R, Cheng X, Lachacz E, Thomas D, McMurray K, Smerage J, Henry NL, Heth J, Chinnaiyan A, Hayes DF, Merajver S. Integrative molecular profiling of breast cancer brain metastasis and patient-derived xenograft organoids from resected breast cancer brain metastases to interrogate and prioritize therapeutic personalized strategies [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr PD9-12.