Loss of E-cadherin leads to Id2-dependent inhibition of cell cycle progression in metastatic lobular breast cancer

TY - JOUR

T1 - Loss of E-cadherin leads to Id2-dependent inhibition of cell cycle progression in metastatic lobular breast cancer

AU - Rätze, Max A K

AU - Koorman, Thijs

AU - Sijnesael, Thijmen

AU - Bassey-Archibong, Blessing

AU - van de Ven, Robert

AU - Enserink, Lotte

AU - Visser, Daan

AU - Jaksani, Sridevi

AU - Viciano, Ignacio

AU - Bakker, Elvira R M

AU - Richard, François

AU - Tutt, Andrew

AU - O'Leary, Lynda

AU - Fitzpatrick, Amanda

AU - Roca-Cusachs, Pere

AU - van Diest, Paul J

AU - Desmedt, Christine

AU - Daniel, Juliet M

AU - Isacke, Clare M

AU - Derksen, Patrick W B

N1 - Funding Information: We thank Jonathan R. Keller (National Cancer Institute, Frederick, MA, USA) for the Id2 reporter, and Livio Kleij and Nikki Kolsters for technical support. Members from the De Bruin and Daniel laboratories are acknowledged for input and fruitful discussions. We thank the UMC Utrecht Cell Microscopy Center for imaging support. Rebecca Marlow and Eleanor Knight (BCN Organoid Facility) for their advice on the ILC PDO generation and Angela Clifford (KCL) in her role as BTBC trial coordinator. We acknowledge NHS funding to the NIHR Biomedical Research Centres at both Guy’s and St Thomas’ NHS Foundation Trust and King’s College London, and the Royal Marsden NHS Foundation Trust and the Canadian Breast Cancer Foundation, the Juravinski Hospital and Cancer Center Foundation (JHCCF). BIB-A was partly supported by a Schlumberger Faculty for the Future Fellowship. We received financial support by Breast Cancer Now (CTR-Q4-Y3), NC3Rs (NC/P001262/1) (CMI/AT) and a Medical Research Council Clinical Research Training Fellowship (AF), grants from the Netherlands Organization for Scientific Research (NWO/ZonMW-VIDI 016.096.318), the Dutch Cancer Society (KWF-UU-2011-5230, KWF-UU-2014-7201 and KWF-UU-2016-10456), Breast Cancer Now (2018NovPCC1297) which is supported by funding from Pfizer, and the European Union’s Horizon 2020 FET Proactive program under the grant agreement No. 731957 (MECHANO-CONTROL). This publication is based upon work from COST action LOBSTERPOT (CA19138), supported by COST (European Cooperation in Science and Technology). Funding Information: We thank Jonathan R. Keller (National Cancer Institute, Frederick, MA, USA) for the Id2 reporter, and Livio Kleij and Nikki Kolsters for technical support. Members from the De Bruin and Daniel laboratories are acknowledged for input and fruitful discussions. We thank the UMC Utrecht Cell Microscopy Center for imaging support. Rebecca Marlow and Eleanor Knight (BCN Organoid Facility) for their advice on the ILC PDO generation and Angela Clifford (KCL) in her role as BTBC trial coordinator. We acknowledge NHS funding to the NIHR Biomedical Research Centres at both Guy’s and St Thomas’ NHS Foundation Trust and King’s College London, and the Royal Marsden NHS Foundation Trust and the Canadian Breast Cancer Foundation, the Juravinski Hospital and Cancer Center Foundation (JHCCF). BIB-A was partly supported by a Schlumberger Faculty for the Future Fellowship. We received financial support by Breast Cancer Now (CTR-Q4-Y3), NC3Rs (NC/P001262/1) (CMI/AT) and a Medical Research Council Clinical Research Training Fellowship (AF), grants from the Netherlands Organization for Scientific Research (NWO/ZonMW-VIDI 016.096.318), the Dutch Cancer Society (KWF-UU-2011-5230, KWF-UU-2014-7201 and KWF-UU-2016-10456), Breast Cancer Now (2018NovPCC1297) which is supported by funding from Pfizer, and the European Union’s Horizon 2020 FET Proactive program under the grant agreement No. 731957 (MECHANO-CONTROL). This publication is based upon work from COST action LOBSTERPOT (CA19138), supported by COST (European Cooperation in Science and Technology). Publisher Copyright: © 2022, The Author(s).

PY - 2022/5/20

Y1 - 2022/5/20

N2 - Invasive lobular breast carcinoma (ILC) is characterized by proliferative indolence and long-term latency relapses. This study aimed to identify how disseminating ILC cells control the balance between quiescence and cell cycle re-entry. In the absence of anchorage, ILC cells undergo a sustained cell cycle arrest in G0/G1 while maintaining viability. From the genes that are upregulated in anchorage independent ILC cells, we selected Inhibitor of DNA binding 2 (Id2), a mediator of cell cycle progression. Using loss-of-function experiments, we demonstrate that Id2 is essential for anchorage independent survival (anoikis resistance) in vitro and lung colonization in mice. Importantly, we find that under anchorage independent conditions, E-cadherin loss promotes expression of Id2 in multiple mouse and (organotypic) human models of ILC, an event that is caused by a direct p120-catenin/Kaiso-dependent transcriptional de-repression of the canonical Kaiso binding sequence TCCTGCNA. Conversely, stable inducible restoration of E-cadherin expression in the ILC cell line SUM44PE inhibits Id2 expression and anoikis resistance. We show evidence that Id2 accumulates in the cytosol, where it induces a sustained and CDK4/6-dependent G0/G1 cell cycle arrest through interaction with hypo-phosphorylated Rb. Finally, we find that Id2 is indeed enriched in ILC when compared to other breast cancers, and confirm cytosolic Id2 protein expression in primary ILC samples. In sum, we have linked mutational inactivation of E-cadherin to direct inhibition of cell cycle progression. Our work indicates that loss of E-cadherin and subsequent expression of Id2 drive indolence and dissemination of ILC. As such, E-cadherin and Id2 are promising candidates to stratify low and intermediate grade invasive breast cancers for the use of clinical cell cycle intervention drugs.

AB - Invasive lobular breast carcinoma (ILC) is characterized by proliferative indolence and long-term latency relapses. This study aimed to identify how disseminating ILC cells control the balance between quiescence and cell cycle re-entry. In the absence of anchorage, ILC cells undergo a sustained cell cycle arrest in G0/G1 while maintaining viability. From the genes that are upregulated in anchorage independent ILC cells, we selected Inhibitor of DNA binding 2 (Id2), a mediator of cell cycle progression. Using loss-of-function experiments, we demonstrate that Id2 is essential for anchorage independent survival (anoikis resistance) in vitro and lung colonization in mice. Importantly, we find that under anchorage independent conditions, E-cadherin loss promotes expression of Id2 in multiple mouse and (organotypic) human models of ILC, an event that is caused by a direct p120-catenin/Kaiso-dependent transcriptional de-repression of the canonical Kaiso binding sequence TCCTGCNA. Conversely, stable inducible restoration of E-cadherin expression in the ILC cell line SUM44PE inhibits Id2 expression and anoikis resistance. We show evidence that Id2 accumulates in the cytosol, where it induces a sustained and CDK4/6-dependent G0/G1 cell cycle arrest through interaction with hypo-phosphorylated Rb. Finally, we find that Id2 is indeed enriched in ILC when compared to other breast cancers, and confirm cytosolic Id2 protein expression in primary ILC samples. In sum, we have linked mutational inactivation of E-cadherin to direct inhibition of cell cycle progression. Our work indicates that loss of E-cadherin and subsequent expression of Id2 drive indolence and dissemination of ILC. As such, E-cadherin and Id2 are promising candidates to stratify low and intermediate grade invasive breast cancers for the use of clinical cell cycle intervention drugs.

UR - http://www.scopus.com/inward/record.url?scp=85128251885&partnerID=8YFLogxK

U2 - 10.1038/s41388-022-02314-w

DO - 10.1038/s41388-022-02314-w

M3 - Article

C2 - 35437308

SN - 0950-9232

VL - 41

SP - 2932

EP - 2944

JO - Oncogene

JF - Oncogene

IS - 21

ER -