Open AccessDevelopment of a Fluorescent Reporter System to Delineate Cancer Stem Cells in Triple‐Negative Breast Cancer
Author(s) -
Thiagarajan Praveena S.,
Hitomi Masahiro,
Hale James S.,
Alvarado Alvaro G.,
Otvos Balint,
Sinyuk Maksim,
Stoltz Kevin,
Wiechert Andrew,
MulkearnsHubert Erin,
Jarrar Awad M.,
Zheng Qiao,
Thomas Dustin,
Egelhoff Thomas T.,
Rich Jeremy N.,
Liu Huiping,
Lathia Justin D.,
Reizes Ofer
Publication year - 2015
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1002/stem.2021
Subject(s) - homeobox protein nanog , sox2 , cancer stem cell , cd44 , biology , triple negative breast cancer , green fluorescent protein , stem cell , cancer research , lin28 , stem cell marker , microbiology and biotechnology , cell , embryonic stem cell , cancer , induced pluripotent stem cell , breast cancer , genetics , gene
Abstract Advanced cancers display cellular heterogeneity driven by self‐renewing, tumorigenic cancer stem cells (CSCs). The use of cell lines to model CSCs is challenging due to the difficulty of identifying and isolating cell populations that possess differences in self‐renewal and tumor initiation. To overcome these barriers in triple‐negative breast cancer (TNBC), we developed a CSC system using a green fluorescent protein (GFP) reporter for the promoter of the well‐established pluripotency gene NANOG . NANOG‐GFP+ cells gave rise to both GFP+ and GFP − cells, and GFP+ cells possessed increased levels of the embryonic stem cell transcription factors NANOG, SOX2, and OCT4 and elevated self‐renewal and tumor initiation capacities. GFP+ cells also expressed mesenchymal markers and demonstrated increased invasion. Compared with the well‐established CSC markers CD24 − /CD44 + , CD49f, and aldehyde dehydrogenase (ALDH) activity, our NANOG‐GFP reporter system demonstrated increased enrichment for CSCs. To explore the utility of this system as a screening platform, we performed a flow cytometry screen that confirmed increased CSC marker expression in the GFP+ population and identified new cell surface markers elevated in TNBC CSCs, including junctional adhesion molecule‐A (JAM‐A). JAM‐A was highly expressed in GFP+ cells and patient‐derived xenograft ALDH+ CSCs compared with the GFP − and ALDH − cells, respectively. Depletion of JAM‐A compromised self‐renewal, whereas JAM‐A overexpression induced self‐renewal in GFP − cells. Our data indicate that we have defined and developed a robust system to monitor differences between CSCs and non‐CSCs in TNBC that can be used to identify CSC‐specific targets for the development of future therapeutic strategies. Stem Cells. S tem C ells 2015;33:2114–2125