Gene signatures of circulating breast cancer cell models are a source of novel molecular determinants of metastasis and improve circulating tumor cell detection in patients

Abstract

Background Progression to stage IV disease remains the main cause of breast cancer-related deaths. Increasing knowledge on the hematogenous phase of metastasis is key for exploiting the entire window of opportunity to interfere with early dissemination and to achieve a more effective disease control. Recent evidence suggests that circulating tumor cells (CTCs) possess diverse adaptive mechanisms to survive in blood and eventually metastasize, encouraging research into CTC-directed therapies. Methods On the hypothesis that the distinguishing molecular features of CTCs reveal useful information on metastasis biology and disease outcome, we compared the transcriptome of CTCs, primary tumors, lymph-node and lung metastases of the MDA-MB-231 xenograft model, and assessed the biological role of a panel of selected genes, by in vitro and in vivo functional assays, and their clinical significance in M0 and M+ breast cancer patients. Results We found that hematogenous dissemination is governed by a transcriptional program and identified a CTC signature that includes 192 up-regulated genes, mainly related to cell plasticity and adaptation, and 282 down-regulated genes, involved in chromatin remodeling and transcription. Among genes up-regulated in CTCs, FADS3 was found to increases cell membrane fluidity and promote hematogenous diffusion and lung metastasis formation. TFF3 was observed to be associated with a subset of CTCs with epithelial-like features in the experimental model and in a cohort of 44 breast cancer patients, and to play a role in cell migration, invasion and blood-borne dissemination. The analysis of clinical samples with a panel of CTC-specific genes (ADPRHL1, ELF3, FCF1, TFF1 and TFF3) considerably improved CTC detection as compared with epithelial and tumor-associated markers both in M0 and stage IV patients, and CTC kinetics informed disease relapse in the neoadjuvant setting. Conclusions Our findings provide evidence on the potential of a CTC-specific molecular profile as source of metastasis-relevant genes in breast cancer experimental models and in patients. Thanks to transcriptome analysis we generated a novel CTC signature in the MDA-MB-231 xenograft model, adding a new piece to the current knowledge on the key players that orchestrate tumor cell hematogenous dissemination and breast cancer metastasis, and expanding the list of CTC-related biomarkers for future validation studies.

Accurate Screening for Early-Stage Breast Cancer by Detection and Profiling of Circulating Tumor Cells

“…Similarly, Jin et al [ 10 ] use the CytoSorter ® CTC capture system, and show 50% and >80% sensitivity in DCIS and stage I/II BrC. Fina et al report >78% CTC detection rates in early-stage breast cancers using an antigen-independent method [ 11 ], and 65% CTC detection rate using antigen-dependent (EpCAM, ERBB2, and EGFR expression) capture, followed by quantitative polymerase chain reaction (qPCR) profiling of targeted gene panel [ 12 ]. These studies support the biological plausibility of CTC-based cancer screening approaches.…”

“…Most prior attempts at evaluating CTCs for cancer screening were based on epitope capture using the CellSearch platform, which, while not approved for CTC detection, is frequently used in research. Several prior studies highlight the lower performance of epitope capture, arising due to its inability to efficiently harvest or detect CTCs with lower expression of EpCAM and PanCK, which are the most routinely employed target markers [ 13 , 14 , 15 , 16 , 17 , 18 , 19 ], with some improvements in sensitivity when epitope capture is used in combination with gene expression profiling [ 12 ]. We previously described a novel functional enrichment method with high CTC detection sensitivity, which yields sufficient CTCs for downstream applications, such as immunocytochemistry (ICC) profiling [ 20 , 21 ].…”

Circulating Tumor Cells and Breast Cancer Metastasis: From Enumeration to Somatic Mutational Profile

“…We reviewed a total of 378 research papers in the PubMed database from 2010 to mid-2022 and observed an increasing trend in the number of publications, with an annual growth rate of 12.25%. We summarized the recent studies on the association between CTCs and breast cancer metastasis in Table 1 [ 14 , 22 , 23 , 24 , 25 , 26 , 27 , 28 ].…”

Landscape of RNA-binding proteins in diagnostic utility, immune cell infiltration and PANoptosis features of heart failure

“…EBNA1BP2 functions as a dynamic scaffold for ribosome biogenesis (Hirano et al, 2009). FCF1 is a potential marker of circulating breast cancer cells for detecting metastasis (Fina et al, 2022). The nucleolar GTPbinding protein GNL2 is essential for retinal neurogenesis in developing zebrafish (Paridaen et al, 2011).…”