Abstract P5-06-01: A Multi-center Clinical Study to Harvest and Characterize Circulating Tumor Cells from Patients with Metastatic Breast Cancer Using the Parsortix® PC1 System in support of FDA clearance

Abstract Background: Circulating tumor cells (CTCs) captured from the blood of cancer patients may serve as a non-invasive surrogate source of tumor material to investigate tumor characteristics in real-time. However, the only FDA-cleared CTC assay is limited to the enumeration of surface marker-defined epithelial cells and not designed for further characterization of the CTCs identified. The Parsortix® PC1 system is a semi-automated microfluidic device capable of capturing and harvesting CTCs from peripheral blood based on cell size and deformability, making it cell-surface marker agnostic. Here, we demonstrate that the Parsortix® PC1 system enables the enrichment and capture of CTCs from the blood of patients with metastatic breast cancer (MBC) and their interrogation using evaluation techniques commonly available in clinical laboratories. Methods: As part of a multicenter clinical trial (NCT03427450), peripheral blood samples from 216 patients with MBC and 205 healthy volunteers (HVs) were prospectively collected at four different clinical sites located throughout the United States. Each subject provided two separate blood samples collected into K2EDTA Vacutainer® tubes to be processed using the Parsortix® PC1 system on the same day. The cells harvested from one of the blood samples collected from each subject by the Parsortix® PC1 system were deposited onto cytology slides using a cytocentrifugation method and stained with Wright-Giemsa reagents using an automated stainer. The stained slides were subjected to cytopathological evaluation by a board-certified pathologist to enumerate CTCs. As proof of principle, cells harvested from the second blood sample were evaluated using one of three additional techniques: molecular profiling by qRT-PCR, RNA sequencing, or cytogenetic analysis of HER2 amplification by FISH. Results: Cytologic examination identified one or more cells as a CTC in 48.5% (95% CI of 41.5 – 55.4%) of the 194 patients with MBC and 9.9% (95% CI of 6.4 – 14.9%) of the 192 HVs. The results from the qRT-PCR evaluation (102 HVs and 74 MBC patients) showed differential expression of cancer-related genes (KRT19, EPCAM, and TWIST1) in the patients with MBC compared to the HVs. Results from the RNA sequencing (53 HVs and 16 MBC patients) showed differential expression of several genes involved in the Kegg Cancer Pathway in the patients with MBC compared to the HVs. The results from the HER2 FISH evaluation (38 HVs and 101 MBC patients) showed that while the majority of the CTC identified had normal HER2/CEP17 ratios, detection of HER2 amplification was possible. Conclusions: The Parsortix PC1 system is capable of capturing and harvesting CTCs from the peripheral blood of patients with MBC. Harvested cells can be evaluated using standard orthogonal methodologies such as gene expression and FISH to identify and characterize CTCs. Based in part on the above results, the FDA granted a De Novo classification request (DEN200062) for the Parsortix PC1 device in May of 2022. Citation Format: Evan Cohen, Gitanjali Jayachandran, Richard Moore, Massimo Cristofanilli, Julie E. Lang, Joseph Khoury, Michael F. Press, Heather McBride, Kyu Kwang Kim, Negar Khazan, Qiang Zhang, Youbin Zhang, Roberta Guzman, Michael C. Miller, James Reuben, Naoto T. Ueno. A Multi-center Clinical Study to Harvest and Characterize Circulating Tumor Cells from Patients with Metastatic Breast Cancer Using the Parsortix® PC1 System in support of FDA clearance [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P5-06-01.