TY - JOUR
T1 - Sleeping beauty genetic screen identifies miR-23b::BTBD7 gene interaction as crucial for colorectal cancer metastasis
AU - Grisard, Eleonora
AU - Coan, Michela
AU - Cesaratto, Laura
AU - Rigo, Ilenia
AU - Zandonà, Luigi
AU - Paulitti, Alice
AU - Andreuzzi, Eva
AU - Rampioni Vinciguerra, Gian Luca
AU - Poletto, Evelina
AU - Del Ben, Fabio
AU - Brisotto, Giulia
AU - Biscontin, Eva
AU - Turetta, Matteo
AU - Dassi, Erik
AU - Mirnezami, Alex
AU - Canzonieri, Vincenzo
AU - Vecchione, Andrea
AU - Baldassarre, Gustavo
AU - Mongiat, Maurizio
AU - Spizzo, Riccardo
AU - Nicoloso, Milena S.
PY - 2019/8
Y1 - 2019/8
N2 - Background: Metastatic colorectal cancer (CRC) remains a deadly disease. Identifying locally advanced CRC patients with high risk of developing metastasis and improving outcome of metastatic CRC patients require discovering master regulators of metastasis. In this context, the non-coding part of the human genome is still largely unexplored. Methods: To interrogate the non-coding part of the human genome and disclose regulators of CRC metastasis, we combined a transposon-based forward genetic screen with a novel in vitro assay, which forces cells to grow deprived of cell-substrate and cell-cell contacts (i.e. forced single cell suspension assay - fSCS). Findings: We proved that fSCS selects CRC cells with mesenchymal and pro-metastatic traits. Moreover, we found that the transposon insertions conferred CRC cells resistance to fSCS and thus metastatic advantage. Among the retrieved transposon insertions, we demonstrated that the one located in the 3′UTR of BTBD7 disrupts miR-23b::BTBD7 interaction and contributes to pro-metastatic traits. In addition, miR-23b and BTBD7 correlate with CRC metastasis both in preclinical experiments and in clinical samples. Interpretation: fSCS is a simple and scalable in vitro assay to investigate pro-metastatic traits and transposon-based genetic screens can interrogate the non-coding part of the human genome (e.g. miRNA::target interactions). Finally, both Btbd7 and miR-23b represent promising prognostic biomarkers and therapeutic targets in CRC. Fund: This work was supported by Marie Curie Actions (CIG n. 303877) and Friuli Venezia Giulia region (Grant Agreement n°245574), Italian Association for Cancer Research (AIRC, MFAG n°13589), Italian Ministry of Health (GR-2010-2319387 and PE-2016-02361040) and 5x1000 to CRO Aviano.
AB - Background: Metastatic colorectal cancer (CRC) remains a deadly disease. Identifying locally advanced CRC patients with high risk of developing metastasis and improving outcome of metastatic CRC patients require discovering master regulators of metastasis. In this context, the non-coding part of the human genome is still largely unexplored. Methods: To interrogate the non-coding part of the human genome and disclose regulators of CRC metastasis, we combined a transposon-based forward genetic screen with a novel in vitro assay, which forces cells to grow deprived of cell-substrate and cell-cell contacts (i.e. forced single cell suspension assay - fSCS). Findings: We proved that fSCS selects CRC cells with mesenchymal and pro-metastatic traits. Moreover, we found that the transposon insertions conferred CRC cells resistance to fSCS and thus metastatic advantage. Among the retrieved transposon insertions, we demonstrated that the one located in the 3′UTR of BTBD7 disrupts miR-23b::BTBD7 interaction and contributes to pro-metastatic traits. In addition, miR-23b and BTBD7 correlate with CRC metastasis both in preclinical experiments and in clinical samples. Interpretation: fSCS is a simple and scalable in vitro assay to investigate pro-metastatic traits and transposon-based genetic screens can interrogate the non-coding part of the human genome (e.g. miRNA::target interactions). Finally, both Btbd7 and miR-23b represent promising prognostic biomarkers and therapeutic targets in CRC. Fund: This work was supported by Marie Curie Actions (CIG n. 303877) and Friuli Venezia Giulia region (Grant Agreement n°245574), Italian Association for Cancer Research (AIRC, MFAG n°13589), Italian Ministry of Health (GR-2010-2319387 and PE-2016-02361040) and 5x1000 to CRO Aviano.
KW - Colorectal cancer metastasis
KW - DNA transposons
KW - microRNA target
KW - spleeping beauty
UR - http://www.scopus.com/inward/record.url?scp=85068580726&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85068580726&partnerID=8YFLogxK
U2 - 10.1016/j.ebiom.2019.06.044
DO - 10.1016/j.ebiom.2019.06.044
M3 - Article
C2 - 31303496
AN - SCOPUS:85068580726
VL - 46
SP - 79
EP - 93
JO - EBioMedicine
JF - EBioMedicine
SN - 2352-3964
ER -