microRNA Expression Profile in Single Hormone Receptor-Positive Breast Cancers Is Mainly Dependent on HER2 Status—A Pilot Study

Abstract

Estrogen (ER) and progesterone (PgR) receptors and HER2 are crucial in the assessment of breast cancer specimens due to their prognostic and predictive significance. Single hormone receptor-positive breast cancers are less common and their clinical course is less favorable than ER(+)/PgR(+) tumors. Their molecular features, especially microRNA (miRNA) profiles, have not been investigated to date. Tumor specimens from 36 chemonaive breast cancer patients with known ER and PgR status (18 ER(+)/PgR(−) and 18 ER(−)/PgR(+) cases) were enrolled to the study. The expression of 829 miRNAs was evaluated with nCounter Human v3 miRNA expression Assay (NanoString). miRNAs differentiating between ER/PgR/HER2 phenotypes were selected based on fold change (FC) calculated for the mean normalized counts of each probe in compared groups. The differences were estimated with Student’s T-test or Two-Way ANOVA (considering also the HER2 status). The results were validated using The Cancer Genome Atlas (TCGA) dataset. Following quality control of raw data, fourcases were excluded due to low sample quality, leaving 14 ER(+)/PgR(−) and 18 ER(−)/PgR(+) cases. After correction for multiple comparisons, we did not find miRNA signature differentiating between ER(−)/PgR(+) and ER(+)/PgR(−) breast cancers. However, a trend for differing expression (p-value ≤ 0.05; FDR > 0.2; ANOVA) in eight miRNAs was observed. The ER(+)/PgR(−) group demonstrated elevated levels of four miRNAs—miR-30a-5p, miR-29c-3p, miR-141-3p and miR-423-5p—while the ER(−)/PgR(+) tumors were enriched in another four miRNAs—miR-514b-5p, miR-424-5p, miR-495-3p, and miR-92a-3p. For one of the miRNAs—miR-29c-3p—the association with the ER(+)/PgR(−) phenotype was confirmed in the TCGA cohort (p-value = 0.024; T-test). HER2 amplification/overexpression in the NanoString cohort was related to significant differences observed in 33 miRNA expression levels (FDR ≤ 0.2; ANOVA). The association with HER2 status was confirmed in the TCGA cohort for four miRNAs (miR-1180-3p, miR-223-3p, miR-30d-5p, and miR-195-5p). The main differences in miRNA expression amongst single hormone receptor-positive tumors were identified according to their HER2 status. However, ER(+)/PgR(−) cases tended to express higher levels of miRNAs associated with ER-positivity (miR-30a-5p, miR-29c-3p, miR-141-3p), whereas ER(−)/PgR(+) cancers showed elevated levels of miRNAs characteristic for double- and triple-negative tumors (miR-92a-3p, miR-424-5p). Further studies are necessary to comprehensively analyze miRNA signatures characteristic of ER(−)/PgR(+) and ER(+)/PgR(−) tumors.

Lost but Not Least—Novel Insights into Progesterone Receptor Loss in Estrogen Receptor-Positive Breast Cancer

“…Importantly, these associations are not seen in HER2(+) samples, suggesting that the effects of HER2 are dominant. This is further supported by our study on single hormone receptor BC, in which miRNA profiles of single hormone receptor-positive breast cancers were mainly dependent on the status of HER2, rather than on ERα/PgR status [83].…”

“…In our recent study, we compared miRNA profiles of two groups of single-steroid-hormone-receptor-positive BC, ER(+)/PgR(−) and ER(−)/PgR(+) [ 83 ]. The first group demonstrated elevated levels of miR-30a-5p, miR-29c-3p, miR-141-3p—members of miRNA clusters characterizing ER(+) tumors, and miR-423-5p, whose role in PgR silencing was discussed before [ 77 ].…”

“…The first group demonstrated elevated levels of miR-30a-5p, miR-29c-3p, miR-141-3p—members of miRNA clusters characterizing ER(+) tumors, and miR-423-5p, whose role in PgR silencing was discussed before [ 77 ]. Interestingly, miR-30-5p has previously been shown to suppress PgR expression in BC cell lines [ 83 ]. Additionally, the miR-29 family targets and represses transcription of the PgR-regulated gene, ATP1B1 [ 82 ].…”

“…This was confirmed also in our analysis of the TCGA dataset, where we identified 2 and 32 differently expressed genes between ER(+)/PgR(−) and double-positive or double negative tumors, respectively. Importantly, we found only 10 genes uniquely differentiating between two subtypes of single hormone receptor-positive tumors [ 83 ].…”

The many facets of miR‐223 in cancer: Oncosuppressor, oncogenic driver, therapeutic target, and biomarker of response

“…As expected, many miRNAs have been deeply involved in the regulation of different pathway in BC, such as those involved in proliferation, apoptosis, metastasis, recurrence, and drug resistance (Loh et al, 2019 ). At this regard, it has been reported that downregulation of miR‐223‐3p, as a part of a 4‐miR‐signature, is associated with HER2 status in the TCGA cohort (Kunc et al, 2020 ).…”