Fig 1.
AR expression by IHC staining.
Representative images of AR-positive (A) and AR-negative (B) tumors. Arrows: tumors; arrowhead, normal breast epithelium. Original magnifications, x100.
Table 1.
Baseline patient characteristics (N = 68).
Fig 2.
Representative images of CTC subtypes identified by the Epic Sciences CTC platform.
Representative fluorescence microscopy images of subtypes of CTCs identified by the Epic Sciences CTC platform. Blood samples from patients with metastatic breast cancer were deposited on glass slides and stained with a cocktail of DAPI and antibodies against CK, CD45, and AR. After staining, CTCs were detected using a digital pathology algorithm and classified into CTC subtypes on the basis of marker expression profile into CK+ CTCs, CK- CTCs, CTC clusters, and apoptotic CTCs. The top panel shows an AR+CK+ CTC with AR expression localized in the nucleus.
Fig 3.
Prevalence of AR+ and ER+ CTCs in metastatic breast cancer samples.
(A) Dot plot depicting AR expression in CTCs identified in patients with metastatic breast cancer with respect to the threshold for AR positivity (indicated by dotted line). Each dot represents a single CTC, and the color indicates the subtype, defined as CK+, CK-, CK+ cluster, or apoptotic (Apop). AR+ CTCs were detected in 10 of 68 patients tested. The 2 patients with only apoptotic AR+ CTCs were excluded. Interpatient heterogeneity was observed in levels of AR expression and subtypes of CTCs. N-term, N-terminal region. (B) Dot plot depicting ER expression in CTCs in the 10 patients with AR+ CTCs with respect to the threshold for ER positivity (indicated by dotted line). Each dot represents a single CTC, and the color indicates the subtype, defined as CK+, CK-, CK+ cluster, or apoptotic (Apop). ER+ CTCs were identified in 3 of 10 patients tested.
Table 2.
Clinicopathological characteristics of the 12 Patients with AR+ CTCs.
Fig 4.
Single-cell CNV analysis of CTCs.
Seventy eight CTCs with various biomarker (AR, ER and HER2) positive and negative and 1 white blood cell (germline control) detected in the sample from patient 11 were sequenced and analyzed for the presence of CNVs. (A) Characteristics of CTCs sequenced for CNV analysis according to AR, ER, and HER2 expression. (B) Representative examples of the 3 different CNV patterns identified in patient #11. The bottom figure is the CNV profile of the WBC. (C) Incidences of the 3 CNV patterns in CTCs according to AR, ER, and HER2 expression.