Charlotte Rypens1,2, Melike Marsan3, Christophe Van Berckelaer4,3,5, Charlotte Billiet4,6, Kirsten Melis7, Sara Perez Lopez8, Peter van Dam4,5, Gayathri R Devi9, Pascal Finetti10, Naoto T Ueno11, François Bertucci10, Piet Dirix4,6, Patrick Neven12, Peter Vermeulen4,3, Luc Dirix3,13, Steven J Van Laere4,3. 1. Center for Oncological Research (CORE), Faculty of Medicine and Health Sciences - University of Antwerp, Campus Drie Eiken - Universiteitsplein 1, 2610, Wilrijk - Antwerp, Belgium. charlotte.rypens@uantwerpen.be. 2. Translational Cancer Research Unit (TCRU), GZA Ziekenhuizen campus Sint-Augustinus, Oosterveldlaan 24, 2610, Wilrijk - Antwerp, Belgium. charlotte.rypens@uantwerpen.be. 3. Translational Cancer Research Unit (TCRU), GZA Ziekenhuizen campus Sint-Augustinus, Oosterveldlaan 24, 2610, Wilrijk - Antwerp, Belgium. 4. Center for Oncological Research (CORE), Faculty of Medicine and Health Sciences - University of Antwerp, Campus Drie Eiken - Universiteitsplein 1, 2610, Wilrijk - Antwerp, Belgium. 5. Multidisciplinary Breast Clinic, Unit Gynaecologic Oncology, Antwerp University Hospital (UZA), Antwerp, Belgium. 6. Radiation Oncology, Iridium Kankernetwerk, Oosterveldlaan 24, 2610, Wilrijk - Antwerp, Belgium. 7. Karel de Grote Hogeschool, Antwerp, Belgium. 8. University Complutense, Madrid, Spain. 9. Division of Surgical Sciences, Department of Surgery, Duke University Medical Sciences, Durham, NC, USA. 10. Predictive Oncology Team, Centre de Recherche en Cancérologie de Marseille (CRCM), Inserm, CNRS, Aix-Marseille Université, Institut Paoli-Calmettes, Marseille, France. 11. Morgan Welch Inflammatory Breast Cancer Research Program and Clinic & Department of Breast Medical Oncology, The University of Texas, MD Anderson Cancer Center, Houston, TX, USA. 12. Department of Oncology, KU Leuven & Department of Gynaecology and Obstetrics, University Hospitals Leuven, Leuven, Belgium. 13. GZA Ziekenhuizen, campus Sint-Augustinus, Oosterveldlaan 24, 2610, Wilrijk - Antwerp, Belgium.
Abstract
PURPOSE: Inflammatory breast cancer (IBC) is an aggressive form of breast cancer with elevated metastatic potential, characterized by tumor emboli in dermal and parenchymal lymph vessels. This study has investigated the hypothesis that TGFβ signaling is implicated in the molecular biology of IBC. METHODS: TGFβ1-induced cell motility and gene expression patterns were investigated in three IBC and three non-IBC (nIBC) cell lines. Tissue samples from IBC and nIBC patients were investigated for the expression of nuclear SMAD2, SMAD3, and SMAD4. SMAD protein levels were related to gene expression data. RESULTS: TGFβ1-induced cell motility was strongly abrogated in IBC cells (P = 0.003). Genes differentially expressed between IBC and nIBC cells post TGFβ1 exposure revealed attenuated expression of SMAD3 transcriptional regulators, but overexpression of MYC target genes in IBC. IBC patient samples demonstrated a near absence of SMAD3 and -4 expression in the primary tumor compared to nIBC patient samples (P < 0.001) and a further reduction of staining intensity in tumor emboli. Integration of gene and protein expression data revealed that a substantial fraction of the IBC signature genes correlated with SMAD3 and these genes are indicative of attenuated SMAD3 signaling in IBC. CONCLUSION: We demonstrate attenuated SMAD3 transcriptional activity and SMAD protein expression in IBC, together with obliterated TGFβ1-induced IBC cell motility. The further reduction of nuclear SMAD expression levels in tumor emboli suggests that the activity of these transcription factors is involved in the metastatic dissemination of IBC cells, possibly by enabling collective invasion after partial EMT.
PURPOSE: Inflammatory breast cancer (IBC) is an aggressive form of breast cancer with elevated metastatic potential, characterized by tumor emboli in dermal and parenchymal lymph vessels. This study has investigated the hypothesis that TGFβ signaling is implicated in the molecular biology of IBC. METHODS: TGFβ1-induced cell motility and gene expression patterns were investigated in three IBC and three non-IBC (nIBC) cell lines. Tissue samples from IBC and nIBC patients were investigated for the expression of nuclear SMAD2, SMAD3, and SMAD4. SMAD protein levels were related to gene expression data. RESULTS: TGFβ1-induced cell motility was strongly abrogated in IBC cells (P = 0.003). Genes differentially expressed between IBC and nIBC cells post TGFβ1 exposure revealed attenuated expression of SMAD3 transcriptional regulators, but overexpression of MYC target genes in IBC. IBC patient samples demonstrated a near absence of SMAD3 and -4 expression in the primary tumor compared to nIBC patient samples (P < 0.001) and a further reduction of staining intensity in tumor emboli. Integration of gene and protein expression data revealed that a substantial fraction of the IBC signature genes correlated with SMAD3 and these genes are indicative of attenuated SMAD3 signaling in IBC. CONCLUSION: We demonstrate attenuated SMAD3 transcriptional activity and SMAD protein expression in IBC, together with obliterated TGFβ1-induced IBC cell motility. The further reduction of nuclear SMAD expression levels in tumor emboli suggests that the activity of these transcription factors is involved in the metastatic dissemination of IBC cells, possibly by enabling collective invasion after partial EMT.
Entities:
Keywords:
Cell motility; Inflammatory breast cancer; SMAD3; TGFβ
Authors: Maeve Kiely; Lap Ah Tse; Hela Koka; Difei Wang; Priscilla Lee; Feng Wang; Cherry Wu; Koon Ho Tsang; Wing-Cheong Chan; Sze Hong Law; Han Zhang; Eric Karlins; Bin Zhu; Amy Hutchinson; Belynda Hicks; Bin Zhu; Xiaohong R Yang Journal: Epigenetics Date: 2020-09-24 Impact factor: 4.528
Authors: C Van Berckelaer; M Van Geyt; S Linders; C Rypens; X B Trinh; W A A Tjalma; S Van Laere; C Colpaert; L Dirix; P A van Dam Journal: Breast Date: 2020-08-17 Impact factor: 4.380
Authors: Charlotte Rypens; François Bertucci; Pascal Finetti; Fredika Robertson; Sandra V Fernandez; Naoto Ueno; Wendy A Woodward; Kenneth Van Golen; Peter Vermeulen; Luc Dirix; Patrice Viens; Daniel Birnbaum; Gayathri R Devi; Massimo Cristofanilli; Steven Van Laere Journal: NPJ Breast Cancer Date: 2022-01-18