Open AccessMonoclonal Antibodies Against Lgr5 Identify Human Colorectal Cancer Stem Cells
Author(s) -
Kemper Kristel,
Prasetyanti Pramudita R.,
De Lau Wim,
Rodermond Hans,
Clevers Hans,
Medema Jan Paul
Publication year - 2012
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.1233
Subject(s) - lgr5 , cancer stem cell , biology , stem cell , stem cell marker , cd44 , cancer research , clonogenic assay , wnt signaling pathway , colorectal cancer , population , metastasis , immunology , cancer , microbiology and biotechnology , cell , medicine , genetics , signal transduction , environmental health
Abstract In colorectal cancer (CRC), a subpopulation of tumor cells, called cancer stem cell (CSC) fraction, is suggested to be responsible for tumor initiation, growth, and metastasis. The search for a reliable marker to identify these CSCs is ongoing as current markers, like CD44 and CD133, are more broadly expressed and therefore are not highly selective and currently also lack function in CSC biology. Here, we analyzed whether the Wnt target Lgr5, which has earlier been identified as a marker for murine intestinal stem cells, could potentially serve as a functional marker for CSCs. Fluorescence‐activated cell sorting‐based detection of Lgr5, using three newly developed antibodies, on primary colorectal tumor cells revealed a clear subpopulation of Epcam + Lgr5 + cells. Similarly, primary CRC‐derived spheroid cultures, known to be enriched for CSCs, contain high levels of Lgr5 + cells, which decrease upon in vitro differentiation of these CSCs. Selection of the Lgr5 high CRC cells identified the clonogenic fraction in vitro as well as the tumorigenic population in vivo. Finally, we confirm that Lgr5 expression is dependent on the Wnt pathway and show that Lgr5 overexpression induces clonogenic growth. We thus provide evidence that Lgr5 is, next to a functional intestinal stem cell marker, a selective marker for human colorectal CSCs. S TEM C ELLS 2012;30:2378–2386