Protein profiling of fine‐needle aspirates reveals subtype‐associated immune signatures and involvement of chemokines in breast cancer

Abstract

There are increasing demands for informative cancer biomarkers, accessible via minimally invasive procedures, both for initial diagnostics and for follow‐up of personalized cancer therapy, including immunotherapy. Fine‐needle aspiration (FNA) biopsy provides ready access to relevant tissue samples; however, the minute amounts of sample require sensitive multiplex molecular analysis to be of clinical biomarker utility. We have applied proximity extension assays (PEA) to analyze 167 proteins in FNA samples from patients with breast cancer (BC; n = 25) and benign lesions (n = 32). We demonstrate that the FNA BC samples could be divided into two main clusters, characterized by differences in expression levels of the estrogen receptor (ER) and the proliferation marker Ki67. This clustering corresponded to some extent to established BC subtypes. Our analysis also revealed several proteins whose expression levels differed between BC and benign lesions (e.g., CA9, GZMB, IL‐6, VEGFA, CXCL11, PDL1, and PCD1), as well as several chemokines correlating with ER and Ki67 status (e.g., CCL4, CCL8, CCL20, CXCL8, CXCL9, and CXCL17). Finally, we also identified three signatures that could predict Ki67 status, ER status, and tumor grade, respectively, based on a small subset of proteins, which was dominated by chemokines. To our knowledge, expression profiles of CCL13 in benign lesions and BC have not previously been described but were shown herein to correlate with proliferation (P = 0.00095), suggesting a role in advanced BC. Given the broad functional range of the proteins analyzed, immune‐related proteins were overrepresented among the observed alterations. Our pilot study supports the emerging role of chemokines in BC progression. Due to the minimally traumatic sampling and clinically important molecular information for therapeutic decisions, this methodology is promising for future immunoscoring and monitoring of treatment efficacy in BC.

Immune related proteins and tumor infiltratingCD8+ lymphocytes in hypopharyngeal cancer in relation to human papillomavirus (HPV) and clinical outcome

“…This expression pattern is very similar to the pattern earlier obtained for tonsillar and base of tongue cancer analyzed with the same Proseek panel 27 . Also in a study on breast cancer, a similar expression pattern with upregulation of CCL2‐4 and 7‐8, CXCL 9‐11, and IL‐6 in cancer vs benign lesions was observed 30,31 . Notably, upregulation of CCL2‐4, 8, and CXCL9‐11 and 13 are all part of a 12‐chemokine gene expression signature associated with lymph node‐like structures in melanoma, colorectal cancer, breast cancer and melanoma, and proposed to indicate a potential for immune therapy 32‐34 .…”

Apoptotic caspases inhibit abscopal responses to radiation and identify a new prognostic biomarker for breast cancer patients

“…49 Differential gene expression analysis of patient subgroups with good versus bad survival (based on 4 independent signatures of apoptotic proficiency) identified 11 genes similarly enriched in either of these groups, 8 of which had previously been attributed an impact on breast cancer progression or patient survival ( Figures 5 and 6). 34,[37][38][39][40][41][42][43] The expression levels of these genes also conveyed prognostic value in patients from the METABRIC dataset ( Figure 6 and Tables 1and 2), with the potential exception of CYP4X1. Of the remaining three genes, LTB failed to impact cancer-specific overall survival in the METABRIC cohort (Tables 1 and 2), while both CLIC6 and SLC7A2 were strongly associated with improved disease outcome ( Figure 6 and Tables 1 and 2), although only SLC7A2 as an independent prognostic biomarker ( Table 2).…”

“…Strikingly, 7 of these 11 genes have previously been attributed an impact on breast cancer progression and patient survival that is in line with our findings. These genes are AGR3, 34 ANKRD30A, 35 BMPR1B, 36,37 STC2 38,39 and SUSD3, 40 which are all indicators of early disease or good prognosis, as well as CXCL9, 41,42 and CALML5, 43 both of which have previously been linked to disease progression or poor prognosis. Conversely, CYP4X1 (which is our hands was enriched in patients with good survival) has previously been positively correlated with tumor grade.…”

The Importance of Breast Adipose Tissue in Breast Cancer

“…Chemokines are cytokines with a chemotactic behavior. They are widely studied in BC and are usually associated with inflammation and tumorigenesis [ 165 , 166 , 167 ]. Zhao et al, 2018, showed that CCL5 secreted by adipose tissue derived stem cells (ADSC) increases BC proliferation [ 168 ].…”