Sleeping beauty genetic screen identifies miR-23b::BTBD7 gene interaction as crucial for colorectal cancer metastasis.

Adaptor Proteins, Signal Transducing / genetics Cell Communication Cell Line, Tumor Cell Proliferation Colorectal Neoplasms / genetics Epithelial-Mesenchymal Transition / genetics Extracellular Matrix / metabolism Gene Expression Regulation, Neoplastic Genetic Testing Humans MicroRNAs / genetics Neoplasm Metastasis Neoplasm Staging RNA Interference

Colorectal cancer metastasis DNA transposons microRNA target spleeping beauty

Journal

EBioMedicine

ISSN: 2352-3964

Titre abrégé: EBioMedicine

Pays: Netherlands

ID NLM: 101647039

Informations de publication

Date de publication:
Aug 2019

Historique:

received: 09 07 2018

revised: 21 06 2019

accepted: 21 06 2019

pubmed: 16 7 2019

medline: 17 1 2020

entrez: 16 7 2019

Statut: ppublish

Résumé

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. 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). 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. 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.

Sections du résumé

BACKGROUND BACKGROUND

METHODS 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 RESULTS

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 CONCLUSIONS

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.

Identifiants

DOI: 10.1016/j.ebiom.2019.06.044 PMID: 31303496 PMC: PMC6710852

pubmed: 31303496

pii: S2352-3964(19)30425-6

doi: 10.1016/j.ebiom.2019.06.044

pmc: PMC6710852

pii:

doi:

Substances chimiques

Adaptor Proteins, Signal Transducing 0

BTBD7 protein, human 0

MIRN23a microRNA, human 0

MicroRNAs 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

Pagination

79-93

Informations de copyright

Références

Biochim Biophys Acta. 1999 Aug 12;1451(1):1-16

pubmed: 10446384

Cancer Res. 2000 Jan 1;60(1):156-63

pubmed: 10646868

Nature. 2001 Feb 15;409(6822):860-921

pubmed: 11237011

Cancer Res. 2001 Feb 15;61(4):1707-16

pubmed: 11245487

Nature. 2004 Aug 26;430(7003):1034-9

pubmed: 15329723

Cancer Biol Ther. 2005 Apr;4(4):365-70

pubmed: 15846061

Cells Tissues Organs. 2005;179(1-2):56-65

pubmed: 15942193

Nat Rev Cancer. 2006 Jun;6(6):449-58

pubmed: 16723991

Gastroenterology. 2007 Feb;132(2):587-600

pubmed: 17258732

Nucleic Acids Res. 2007 Jul;35(Web Server issue):W71-4

pubmed: 17485472

Methods Mol Biol. 2008;435:95-108

pubmed: 18370070

Cell. 2008 May 16;133(4):704-15

pubmed: 18485877

Mol Biol Cell. 2009 Feb;20(3):1020-9

pubmed: 19056678

Nat Genet. 2009 Jun;41(6):753-61

pubmed: 19412179

Nat Methods. 2009 Jun;6(6):415-22

pubmed: 19478801

J Clin Invest. 2009 Jun;119(6):1420-8

pubmed: 19487818

Cell. 2009 Aug 21;138(4):645-659

pubmed: 19682730

N Engl J Med. 2009 Dec 17;361(25):2449-60

pubmed: 20018966

Cancer Res. 2010 Apr 1;70(7):2789-98

pubmed: 20332227

J Am Coll Surg. 2010 May;210(5):744-52, 752-5

pubmed: 20421043

Science. 2010 Jul 30;329(5991):562-5

pubmed: 20671187

Nat Rev Cancer. 2010 Oct;10(10):696-706

pubmed: 20844553

Mob DNA. 2010 Dec 07;1(1):25

pubmed: 21138556

J Cell Sci. 2011 Oct 1;124(Pt 19):3189-97

pubmed: 21940791

Cell. 2011 Oct 14;147(2):275-92

pubmed: 22000009

Nat Commun. 2011 Nov 22;2:554

pubmed: 22109528

Oncogene. 2012 Oct 25;31(43):4577-87

pubmed: 22266873

Nat Rev Cancer. 2012 May 11;12(6):425-36

pubmed: 22576165

Nat Methods. 2012 Jun 28;9(7):676-82

pubmed: 22743772

Nature. 2012 Jul 18;487(7407):330-7

pubmed: 22810696

Hepatology. 2013 Jun;57(6):2326-37

pubmed: 23325674

J Cell Sci. 2013 Jan 1;126(Pt 1):21-9

pubmed: 23516327

Nat Protoc. 2013 May;8(5):870-91

pubmed: 23558786

Nucleic Acids Res. 2013 May 1;41(10):5400-12

pubmed: 23580553

BMC Med Ethics. 2013 Jul 30;14:30

pubmed: 23899250

Nat Methods. 2013 Oct;10(10):965-71

pubmed: 24161985

Cell Death Dis. 2013 Nov 07;4:e915

pubmed: 24201814

Oncology (Williston Park). 2013 Nov;27(11):1074-8

pubmed: 24575534

Sci Transl Med. 2014 Jun 11;6(240):240ps7

pubmed: 24920658

BMC Cancer. 2014 Sep 24;14:704

pubmed: 25253020

Front Bioeng Biotechnol. 2014 Sep 29;2:38

pubmed: 25325056

Nat Genet. 2015 Jan;47(1):47-56

pubmed: 25485836

Ecancermedicalscience. 2015 Apr 09;9:520

pubmed: 25932044

Nature. 2015 Nov 26;527(7579):472-6

pubmed: 26560033

Nat Biotechnol. 2016 Jun;34(6):631-3

pubmed: 27111720

Cell. 2016 Jun 30;166(1):21-45

pubmed: 27368099

Nat Biotechnol. 2017 Jun;35(6):561-568

pubmed: 28369033

Int Rev Cell Mol Biol. 2017;333:173-228

pubmed: 28729025

Sci Data. 2017 Aug 29;4:170112

pubmed: 28850106

Nat Rev Cancer. 2018 Feb;18(2):128-134

pubmed: 29326430

Nat Cell Biol. 2018 Feb;20(2):211-221

pubmed: 29358704

Nucleic Acids Res. 2019 Jan 8;47(D1):D766-D773

pubmed: 30357393

Nucleic Acids Res. 2019 Jan 8;47(D1):D506-D515

pubmed: 30395287

Cell. 1997 Nov 14;91(4):501-10

pubmed: 9390559