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Heterogeneous atypical cell populations are present in blood of metastatic breast cancer patients.

ABSTRACT: Circulating tumor cells (CTCs) are commonly isolated from the blood by targeting the epithelial cell adhesion molecule (EpCAM) through positive selection. However, EpCAM can be downregulated during metastatic progression, or it can be initially not present. We designed the present prospective trial to characterize CTCs as well as other circulating cell populations in blood samples from women with metastatic breast cancer without EpCAM-dependent enrichment and/or isolation technology.A total of 32 patients with metastatic breast cancer were enrolled, and blood samples were processed using a previously described negative depletion immunomagnetic methodology. Samples from healthy volunteers were run as controls (n?=?5). Multistep sequential labeling was performed to label and fix cell-surface markers followed by permeabilization for cytokeratins (CK) 8, 18 and 19. Multiparametric flow cytometry (FCM) analysis was conducted using a BD LSR II flow cytometer or a BD FACSAria II or FACSAria III cell sorter. Immunocytochemical staining on postenrichment specimens for DAPI, EpCAM, CD45, CK, epidermal growth factor receptor and vimentin was performed. Expression of these markers was visualized using confocal microscopy (CM).CD45-negative/CK-positive (CD45- CK+) populations with EpCAM?+?and EpCAM?-?expression were identified with both FCM and CM from the negatively enriched patient samples. In addition, EpCAM?+?and EpCAM?-?populations that were CK?+?and coexpressing the pan-hematopoietic marker CD45 were also noted. There were more CK?+?EpCAM?-?events/ml than CK?+?EpCAM?+?events/ml in both the CD45- and CD45+ fractions (both statistically significant at P???0.0005). The number of CK?+?CD45- and CK?+?CD45+ events per milliliter in blood samples (regardless of EpCAM status) was higher in patient samples than in normal control samples (P???0.0005 and P???0.026, respectively). Further, a significant fraction of the CK?+?CD45+ events also expressed CD68, a marker associated with tumor-associated macrophages. Higher levels of CD45-CK?+?EpCAM?-?were associated with worse overall survival (P?=?0.0292).Metastatic breast cancer patients have atypical cells that are CK?+?EpCAM?-?circulating in their blood. Because a substantial number of these patients do not have EpCAM?+?CTCs, additional studies are needed to evaluate the role of EpCAM?-?circulating cells as a prognostic and predictive marker.

SUBMITTER: Lustberg MB

PROVIDER: S-EPMC4053256 | biostudies-literature | 2014 Mar

REPOSITORIES: biostudies-literature

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Heterogeneous atypical cell populations are present in blood of metastatic breast cancer patients.

Lustberg Maryam B MB Balasubramanian Priya P Miller Brandon B Garcia-Villa Alejandra A Deighan Clayton C Wu Yongqi Y Carothers Sarah S Berger Michael M Ramaswamy Bhuvaneswari B Macrae Erin R ER Wesolowski Robert R Layman Rachel M RM Mrozek Ewa E Pan Xueliang X Summers Thomas A TA Shapiro Charles L CL Chalmers Jeffrey J JJ

Breast cancer research : BCR 20140306 2

<h4>Introduction</h4>Circulating tumor cells (CTCs) are commonly isolated from the blood by targeting the epithelial cell adhesion molecule (EpCAM) through positive selection. However, EpCAM can be downregulated during metastatic progression, or it can be initially not present. We designed the present prospective trial to characterize CTCs as well as other circulating cell populations in blood samples from women with metastatic breast cancer without EpCAM-dependent enrichment and/or isolation te ...[more]

PMID: 24602188

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Project description:The importance and role of the estrogen receptor (ER) pathway have been well-recognized in both breast cancer development and progression. The treatment of choice in women with estrogen receptor-positive metastatic breast cancer (ER+ mBC) is classically divided into a variety of endocrine therapies, with three of the most common being: selective estrogen receptor modulators (SERM), aromatase inhibitors (AI), and selective estrogen receptor degraders (SERD). However, resistance develops in 30-50% of patients treated with these endocrine therapies due to a sophisticated and at times redundant interference at the molecular level between the ER, growth factors, and downstream cell-signaling pathways. Tumor response is heightened with adjunctive therapy that includes an mTORC1 inhibitor (everolimus), CDK4/6 inhibitors (palbociclib/ribociclib/abemaciclib), and an α isoform-specific PI3K inhibitor (alpelisib). Each of these inhibitors elicits potent anti-proliferative benefits; however, they fail to induce tumor cell death. Consequently, disease progression almost invariably occurs. Evasion of apoptosis is a hallmark of cancer. The p53 and BCL2 represent two important nodes of the apoptosis signaling pathway. Venetoclax, a potent and selective BCL2 inhibitor, synergizes with hormonal therapy in ER+ breast cancer models and is active in clinical trials. Similarly, an MDM2 inhibitor, AMG-232, which induces p53 is active in early clinical trials of both liquid and advanced solid tumor patients. In our ER+ BC cohort (Avera Cancer Institute, Sioux Falls, SD), we observed more than 70% of wild type TP53 and over 10% amplification of MDM2 and MDM4 as comparable with the TCGA data set. We summarized current treatment options, the molecular mechanisms that predispose to endocrine resistance, and a future pro-apoptotic treatment strategy for ER+ mBC patients. Our review presents critical analyses of the therapeutic options for the clinical management of ER+ Metastatic Breast Cancer in the light of a hypothesis targeting the induction of apoptosis in p53 wild type tumors. We reviewed not only the FDA approved current treatment approaches but also presented a discourse addressing the possibilities for novel combination strategy that can induce tumor cell apoptosis, a critical cellular mechanism delaying/denying tumor progression. Our review is unique as it presents patient data in support of our hypothesis.

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Heterogeneous atypical cell populations are present in blood of metastatic breast cancer patients.

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Heterogeneous atypical cell populations are present in blood of metastatic breast cancer patients.

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