TY - JOUR
T1 - NR2F1 Is a Barrier to Dissemination of Early-Stage Breast Cancer Cells
AU - Rodriguez-Tirado, Carolina
AU - Kale, Nupura
AU - Carlini, Maria J.
AU - Shrivastava, Nitisha
AU - Rodrigues, Alcina A.
AU - Khalil, Bassem D.
AU - Bravo-Cordero, Jose Javier
AU - Hong, Yan
AU - Alexander, Melissa
AU - Ji, Jiayi
AU - Behbod, Fariba
AU - Sosa, Maria Soledad
N1 - Funding Information:
The authors thank Dr. Roger Davis for MKK3–/–/MKK6+/– samples and his valuable feedback. They also thank Dr. Skobe for providing the BT474 and MDA-MB-361 cells. Grant support: grant number CCR17483357 (to M.S Sosa); grant number MRF-CDA (to M.S Sosa); grant number K22CA201054 (to M.S. Sosa); CSBC Pilot Project-Sage Bionetworks (to M.S. Sosa); Schneider-Lesser Foundation Fellow Award (to M.S. Sosa); MRA (to M.S. Sosa); BCA (to M.S. Sosa); grant number CCR18547848 (to J.J. Bravo-Cordero); grant number R01CA244780 (to J.J. Bravo-Cordero); grant number K22CA196750 (J.J. Bravo-Cordero); grant number P30-CA196521 (to J.J. Bravo-Cordero); grant number NIH-R00 CA127462 (to F. Behbod); grant number NIH-R01CA207445 (to F. Behbod); grant number NIH-R21CA226567 (to F. Behbod); grant number P30 CA168524 (to F. Behbod); grant number P20 GM130423 (to F, Behbod); grant number UL1TR002366 (to F. Behbod); grant number C38317/A24043 (to F. Behbod); grant number P30CA196521–07 (Icahn School of Medicine at Mount Sinai, ISMMS); grant number 1S10RR026639 (ISMMS).
Funding Information:
Three-dimensional (3D) acini cultures were time-lapse imaged using an inverted microscope Olympus IX-70 with Live Cell enclosed chamber. We recorded four positions per condition in parallel for 2 hours with a 10-minute interval. Temperature was maintained at 37°C and CO2 at 5% throughout the imaging session. Movies were made by using either Metamorph (Molecular Devices) or the open source imaging software ImageJ (RRID:SCR_003070; ref. 18). This was done using the services of the Microscopy CoRE at Icahn School of Medicine at Mount Sinai. This research was supported in part by the Tisch Cancer Institute at Mount Sinai P30 CA196521–Cancer Center Support Grant. Protrusion number and their length-over-width (L/W) ratio were quantified. In L/W ratio plots, data points represent the mean ± SD of measurements taken by two independent operators. Invasive phenotype (a single elongated cell or cell file columns invading the Matrigel) or noninvasive phenotype was determined.
Funding Information:
J.J. Bravo-Cordero reports grants from NCI (grant nos. R01CA244780 and P30-CA196521) and grants from Susan G. Komen/CCR18547848 during the conduct of the study. F. Behbod reports grants from Cancer Research UK and KWF Kanker-bestrijding (reference no. C38317/A24043), grants from NIH (grant nos. NIH-R00 CA127462, NIH-R01CA207445, and NIH-R21CA226567); KU Cancer Center Support Grant (grant no. P30 CA168524); grants from The Kansas Institute for Precision Medicine - COBRE (grant no. P20 GM130423; AKG) during the conduct of the study; and Pilot Grant: NCATS Frontiers-CTSA grant from NCATS awarded to the University of Kansas for Frontiers (reference no. UL1TR002366). M.S. Sosa reports grants, personal fees, and other support from Susan G. Komen CCR17483357, K22 NIH K22CA201054, Melanoma Research Foundation, Melanoma Research Alliance, Schneider-Lesser Foundation Fellow Award, CSBC Pilot Project-Sage Bionetworks, Breast Cancer Alliance; grants and other support from P30CA196521-07; grants and other support from 1S10RR026639 during the conduct of the study; and nonfinancial support from HiberCell LLC outside the submitted work. No disclosures were reported by the other authors.
Funding Information:
Statistical analysis was done using Prism Software (RRID: SCR_002798). Differences were considered significant if P values were <0.05. For the majority of in vitro experiments, two-tailed Student t test was performed unless otherwise specified. For mouse experiments and human samples two-tailed Mann–Whitney test was used. Sample size chosen was done empirically. To determine the frequency of samples carrying NR2F1LOW/PRRX1HIGH ratio between benign adjacent and DCIS samples, Fisher exact test was used. Research reported in this publication was supported in part by the NCI Cancer Center Support Grant P30CA196521–07 awarded to the Tisch Cancer Institute of the Icahn School of Medicine at Mount Sinai and used the Biostatistics Shared Resource Facility. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.
Publisher Copyright:
© 2022 American Association for Cancer Research
PY - 2022/6/15
Y1 - 2022/6/15
N2 - Cancer cells can disseminate during very early and sometimes asymptomatic stages of tumor progression. Though biological barriers to tumorigenesis have been identified and characterized, the mechanisms that limit early dissemination remain largely unknown. We report here that the orphan nuclear receptor nuclear receptor subfamily 2, group F, member 1 (NR2F1)/COUP-TF1 serves as a barrier to early dissemination. NR2F1 expression was decreased in patient ductal carcinoma in situ (DCIS) samples. High-resolution intravital imaging of HER2þ early-stage cancer cells revealed that loss of function of NR2F1 increased in vivo dissemination and was accompanied by decreased E-cadherin expression, activation of wingless-type MMTV integration site family, member 1 (WNT)dependent b-catenin signaling, disorganized laminin 5 deposition, and increased expression of epithelial–mesenchymal transition (EMT) genes such as twist basic helix-loop-helix transcription factor 1 (TWIST1), zinc finger E-box binding homeobox 1 (ZEB1), and paired related homeobox 1 (PRRX1). Furthermore, downregulation of NR2F1 promoted a hybrid luminal/basal phenotype. NR2F1 expression was positively regulated by p38a signaling and repressed by HER2 and WNT4 pathways. Finally, early cancer cells with NR2F1LOW/PRRX1HIGH staining were observed in DCIS samples. Together, these findings reveal the existence of an inhibitory mechanism of dissemination regulated by NR2F1 in early-stage breast cancer cells. Significance: During early stages of breast cancer progression, HER2-mediated suppression of NR2F1 promotes dissemination by inducing EMT and a hybrid luminal/basal-like program.
AB - Cancer cells can disseminate during very early and sometimes asymptomatic stages of tumor progression. Though biological barriers to tumorigenesis have been identified and characterized, the mechanisms that limit early dissemination remain largely unknown. We report here that the orphan nuclear receptor nuclear receptor subfamily 2, group F, member 1 (NR2F1)/COUP-TF1 serves as a barrier to early dissemination. NR2F1 expression was decreased in patient ductal carcinoma in situ (DCIS) samples. High-resolution intravital imaging of HER2þ early-stage cancer cells revealed that loss of function of NR2F1 increased in vivo dissemination and was accompanied by decreased E-cadherin expression, activation of wingless-type MMTV integration site family, member 1 (WNT)dependent b-catenin signaling, disorganized laminin 5 deposition, and increased expression of epithelial–mesenchymal transition (EMT) genes such as twist basic helix-loop-helix transcription factor 1 (TWIST1), zinc finger E-box binding homeobox 1 (ZEB1), and paired related homeobox 1 (PRRX1). Furthermore, downregulation of NR2F1 promoted a hybrid luminal/basal phenotype. NR2F1 expression was positively regulated by p38a signaling and repressed by HER2 and WNT4 pathways. Finally, early cancer cells with NR2F1LOW/PRRX1HIGH staining were observed in DCIS samples. Together, these findings reveal the existence of an inhibitory mechanism of dissemination regulated by NR2F1 in early-stage breast cancer cells. Significance: During early stages of breast cancer progression, HER2-mediated suppression of NR2F1 promotes dissemination by inducing EMT and a hybrid luminal/basal-like program.
UR - http://www.scopus.com/inward/record.url?scp=85132049820&partnerID=8YFLogxK
U2 - 10.1158/0008-5472.CAN-21-4145
DO - 10.1158/0008-5472.CAN-21-4145
M3 - Article
C2 - 35471456
AN - SCOPUS:85132049820
SN - 0008-5472
VL - 82
SP - 2313
EP - 2326
JO - Cancer Research
JF - Cancer Research
IS - 12
ER -