Abstract P2-01-11: Single-cell sequencing of the blood T cell repertoire before and after trastuzumab treatment in early stage HER2+ breast cancer
Authors: Vivian John , Walter Kimberly , Drabek Elliott , Haaser Nicole , Levin Maren K. , Rodriguez Esther San Roman , Peck Kendra , Nguyen Ngan , Millward Carl , Benjamin Jonathan , Robinson William H. , O'Shaughnessy Joyce A. 
Published: March 2022

Abstract
Background: Human epidermal growth factor receptor 2 (HER2) positivity in breast cancer portends an aggressive tumor behavior and poor prognosis and is associated with a positive response to trastuzumab treatment. Prior immunohistochemistry and RNA sequencing of breast tumor tissues suggest that trastuzumab may recruit and activate anti-tumor T cells. Tumor infiltrating lymphocytes have been associated with improved response in patients with HER2+ early breast cancer treated with neoadjuvant trastuzumab plus chemotherapy. However, these cells have not previously been characterized at the single cell level in tumor tissue or in the periphery. Assessing the T cell component in the peripheral blood via single-cell sequencing enables high sensitivity detection of rare cells, may identify T cell antigen receptors (TCR) involved in the anti-tumor response, and could lead to a non-invasive means of monitoring trastuzumab-mediated immune activity. Here we perform single cell sequencing of the blood T cell repertoire in breast cancer patients pre- and post-trastuzumab treatment. Methods: To characterize T cell response in trastuzumab plus chemotherapy treated patients, we profiled peripheral CD3+ T cells using 10x Genomics VDJ single-cell sequencing in paired samples from five patients with HER2+ breast cancer. Patients had stage IIA (n=2), stage IIIC (n=2) or stage IV (n=1) breast cancer and were treated with preoperative docetaxel, carboplatin, trastuzumab, pertuzumab (TCHP). Two patients had a pathological complete response (pCR), and three patients had partial clinical response with residual disease at surgery. Peripheral blood mononuclear cells (PMBCs) were collected at a C1D1 pre-treatment and day 1 of cycles 3, 4, or 5. Results: Eleven T cell subpopulations, including naïve and memory CD4+ and CD8+ T cells, activated CD4+ and CD8+ T effector cells, activated CD4+ T regulatory cells, were characterized in the five patients’ peripheral blood based on their transcriptional profiles. T cell subpopulation distribution and clonal expansion profiles were similar in pre- and post- treatment samples in all five donors. Large T cell clonal expansions were detected in the peripheral blood of the two patients who had a pCR, but were not detected in the three patients who had residual disease at surgery. The patients who had a pCR exhibited large expansions in activated CD8+ terminal effector memory/effector memory (TEM/EM) cells followed by expansions in activated CD4+ TEM/EM cells. A minor increasing trend in activated CD4+ Treg cells was observed across all patients upon treatment with TCHP. Conclusions: Single-cell sequencing enables high-resolution characterization of immune cell subsets and represents a promising approach to assess the immune response to trastuzumab and other cancer therapeutics. In this study, single-cell sequencing of peripheral CD3+ T cells identified clonal expansions in activated CD8+ and CD4+ T cells in HER2+ breast cancer patients who had a pCR with preoperative TCHP treatment. These data are consistent with the model that T cells play a key role in trastuzumab-mediated tumor control, and warrant further investigation in a larger sample population.
Citation Format: John Vivian, Kimberly Walter, Elliott Drabek, Nicole Haaser, Maren K. Levin, Esther San Roman Rodriguez, Kendra Peck, Ngan Nguyen, Carl Millward, Jonathan Benjamin, William H. Robinson, Joyce A. O'Shaughnessy. Single-cell sequencing of the blood T cell repertoire before and after trastuzumab treatment in early stage HER2+ breast cancer [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P2-01-11.