Abstract P4-05-20: Case report - 16 year life history and genomic evolution of an ER+ HER2- breast cancer
Authors: Xu Bing , Amallraja Anu , Swaminathan Padmapriya , Elsey Rachel , Davis Christel , Theel Stephanie , Viet Sarah , Petersen Jason , Krie Amy , Davies Gareth E , Williams Casey B , Ehli Erik , Meissner Tobias 
Published: March 2020

Abstract
Introduction
Metastasis is the primary cause of cancer-related death, with genomic evolution between primary and metastatic breast cancer only recently being studied in smaller numbers. We reconstructed the tumor evolution of a female patient originally diagnosed with localized ER+ HER2- disease to investigate the molecular changes that triggered relapse after long term aromatase inhibitor (AI) therapy and tracked them during subsequent treatments. The patient was diagnosed in 2002, relapsed in 2012 (regional lymph node), progressed in 2015 (liver metastasis), and passed away in 2018. We performed DNA- and RNA-sequencing on FFPE tumor samples and multiple liquid biopsies. The patient underwent eight lines of therapy including genome guided targeted therapy.
Materials and Methods
Whole exome and transcriptome sequencing was performed on the patient's primary tumor site at diagnosis (T1) and a metastatic lymph node (M1). Targeted DNA sequencing was performed on a metastatic liver sample (M2, 315 genes) and on liquid biopsies (n=11, 70 genes) every 2-3 months from 2015-2018.
Results
The mutational landscape between timepoints T1, M1 and M2 revealed low mutational burden at diagnosis which ultimately doubled at progression with active mutational processes (APOBEC, POLE) showing concordance between timepoints. DNA-based analysis revealed multiple mutations in the PI3K-Akt signaling pathway. Two mutations were shared between the samples: a clonal PIK3CA Q546R and a TP53 E180K mutation detected as clonal in M1/M2 and subclonal in T1. Individual samples carried private driver mutations, ranging from n=4 in T1, n=7 in M1 to n=16 in M2. While T1 harbored a single clonal driver mutation, with additional driver mutations being subclonal, M1 and M2 harbored multiple clonal driver mutations and a smaller percentage of subclonal driver mutations. A phylogenetic tree showed branching off between T1 and M1/M2. Liquid biopsies, while initially negative, started to detect mutations found among T1, M1 and M2, with increasing allele frequency (AF) throughout time. By the end of 2016, an ESR1 E380Q mutation not previously detected was found at a low AF that had rapidly increased by the end of 2017. RNA-Seq analysis revealed 908 (209 up, 699 down) genes to be differentially expressed between T1 and M1. Most significantly downregulated genes in M1 were TFF1 and PGR indicating loss-of-sensitivity / resistance to AI therapy. Most upregulated genes in M1 included PTHLH, S100P, and SOX2 which have been implicated in promoting tumor growth and metastasis. Pathway analysis between T1 and M1 showed enrichment for the hallmark gene sets ‘Estrogen Response early and late’ and ‘Epithelial mesenchymal transition’.
Conclusions
This phylogenetic reconstruction of the life history of a single patient's cancer revealed an increased mutational rate over time between the patient's primary tumor at diagnosis and samples taken at relapse; and a high level of heterogeneity with metastatic sites sharing only one driver mutation with the primary tumor at diagnosis and having acquired multiple de-novo driver mutations. Transcriptome profiling aided in uncovering the mechanisms that lead to the patient's initial relapse, indicating expression profile changes that promote tumor progression and metastasis as well as reduced sensitivity / resistance to the patients AI based therapy. Monitoring tumor progression through frequent liquid biopsies allowed for the detection of mutations from multiple metastatic sites, in addition to the detection and growth of an AI resistant clone harboring an ESR1 E380Q mutation that was not present in any of the solid tissue samples.
Citation Format: Bing Xu, Anu Amallraja, Padmapriya Swaminathan, Rachel Elsey, Christel Davis, Stephanie Theel, Sarah Viet, Jason Petersen, Amy Krie, Gareth E Davies, Casey B Williams, Erik Ehli, Tobias Meissner. Case report - 16 year life history and genomic evolution of an ER+ HER2- breast cancer [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P4-05-20.