Multiplex imaging reveals novel patterns of MRTFA/B activation in the breast cancer microenvironment

Abstract Background Breast cancer progression and metastasis involve the action of multiple transcription factors in tumors and in the cells of the tumor microenvironment (TME) and understanding how these transcription factors are coordinated can guide novel therapeutic strategies. Myocardin-related...

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Main Authors: Stephanie M. Wilk, Kihak Lee, Caitlyn C. Castillo, Mohamed Haloul, Alexa M. Gajda, Virgilia Macias, Elizabeth L. Wiley, Zhengjia Chen, Xinyi Liu, Xiaowei Wang, Maria Sverdlov, Kent F. Hoskins, Ekrem Emrah Er
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Language:English
Published: BMC 2025-05-01
Series:Journal of Translational Medicine
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Online Access:https://doi.org/10.1186/s12967-025-06559-3
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author Stephanie M. Wilk
Kihak Lee
Caitlyn C. Castillo
Mohamed Haloul
Alexa M. Gajda
Virgilia Macias
Elizabeth L. Wiley
Zhengjia Chen
Xinyi Liu
Xiaowei Wang
Maria Sverdlov
Kent F. Hoskins
Ekrem Emrah Er
author_facet Stephanie M. Wilk
Kihak Lee
Caitlyn C. Castillo
Mohamed Haloul
Alexa M. Gajda
Virgilia Macias
Elizabeth L. Wiley
Zhengjia Chen
Xinyi Liu
Xiaowei Wang
Maria Sverdlov
Kent F. Hoskins
Ekrem Emrah Er
author_sort Stephanie M. Wilk
collection DOAJ
description Abstract Background Breast cancer progression and metastasis involve the action of multiple transcription factors in tumors and in the cells of the tumor microenvironment (TME) and understanding how these transcription factors are coordinated can guide novel therapeutic strategies. Myocardin-related transcription factors A and B (MRTFA/B also known as MKL1/2) are two related transcription factors that redundantly control cancer cell invasion and metastasis in mouse models of breast cancer, but their roles in human cancer are incompletely understood. Here, we investigated the expression and activation of these transcription factors to better assess their tumorigenic and metastatic impact on breast cancer and cells of the tumor microenvironment. Methods We used a multiplexed immunofluorescence approach to label MRTFA, MRTFB, tumor cells by using pan Cytokeratin, endothelial cells by using CD31, and antigen presenting cells (APCs) by using HLA-DRA on two different breast cancer tissue microarrays (TMA): The breast cancer progression TMA provided by the Cooperative Human Tissue Network (CHTN_BrCaProg3) and the University of Illinois Breast Cancer Working Group (TMA BCWG UIC-001-TMA) that included primary tumor and lymph node metastases from patients residing in the West Side and South Side of Chicago. We also used bioinformatics analyses of the TCGA and METABRIC databases and the Broad Institute’s single-cell RNA sequencing portal to investigate MRTFA/B expression patterns in the cells of the tumor microenvironment (TME). Results We found that in human tumors, MRTFA/B are concurrently activated in cancer cells, but they show distinct patterns of expression across different histological subtypes and in the cells of the TME. Importantly, MRTFA expression was elevated in metastatic tumors of African American patients, who disproportionately die from breast cancer. Interestingly, in contrast to publicly available mRNA expression data, MRTFA was similarly expressed across estrogen receptor (ER) positive and negative breast tumors, while MRTFB expression was highest in ER+ breast tumors. Furthermore, MRTFA was specifically expressed in the perivascular antigen-presenting cells (APCs), which has been previously associated with immune suppression and breast cancer progression. We also found that MRTFA expression correlated with the expression of the immune checkpoint protein V-set immunoregulatory receptor (VSIR) in the TCGA data and found that MRTFA activity promotes VSIR expression in THP-1 monocytes and cultured HEK293 cells. Conclusions Our results provide unique insights into how MRTFA and MRTFB promote metastasis in human cancer, the differences of their expression patterns, and their immune suppressive function within the breast cancer TME. Our results will guide future studies on targeting MRTFA/B transcriptional activity and the resulting immune suppression in breast cancer. Graphical Abstract
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spelling doaj-art-bdfa5973ba3c4e13900b580fa1c441f72025-08-20T02:00:03ZengBMCJournal of Translational Medicine1479-58762025-05-0123111910.1186/s12967-025-06559-3Multiplex imaging reveals novel patterns of MRTFA/B activation in the breast cancer microenvironmentStephanie M. Wilk0Kihak Lee1Caitlyn C. Castillo2Mohamed Haloul3Alexa M. Gajda4Virgilia Macias5Elizabeth L. Wiley6Zhengjia Chen7Xinyi Liu8Xiaowei Wang9Maria Sverdlov10Kent F. Hoskins11Ekrem Emrah Er12Department of Physiology and Biophysics, College of Medicine, University of Illinois ChicagoDepartment of Physiology and Biophysics, College of Medicine, University of Illinois ChicagoDepartment of Physiology and Biophysics, College of Medicine, University of Illinois ChicagoDepartment of Physiology and Biophysics, College of Medicine, University of Illinois ChicagoDepartment of Physiology and Biophysics, College of Medicine, University of Illinois ChicagoDepartment of Pathology, University of Illinois ChicagoDepartment of Pathology, University of Illinois ChicagoDivision of Epidemiology and Biostatistics, School of Public Health, University of Illinois ChicagoDepartment of Pharmacology & Regenerative Medicine, College of Medicine, University of Illinois ChicagoDepartment of Pharmacology & Regenerative Medicine, College of Medicine, University of Illinois ChicagoResearch Histology Core, Research Resources Center, University of Illinois ChicagoDivision of Hematology/Oncology, College of Medicine, University of Illinois ChicagoDepartment of Physiology and Biophysics, College of Medicine, University of Illinois ChicagoAbstract Background Breast cancer progression and metastasis involve the action of multiple transcription factors in tumors and in the cells of the tumor microenvironment (TME) and understanding how these transcription factors are coordinated can guide novel therapeutic strategies. Myocardin-related transcription factors A and B (MRTFA/B also known as MKL1/2) are two related transcription factors that redundantly control cancer cell invasion and metastasis in mouse models of breast cancer, but their roles in human cancer are incompletely understood. Here, we investigated the expression and activation of these transcription factors to better assess their tumorigenic and metastatic impact on breast cancer and cells of the tumor microenvironment. Methods We used a multiplexed immunofluorescence approach to label MRTFA, MRTFB, tumor cells by using pan Cytokeratin, endothelial cells by using CD31, and antigen presenting cells (APCs) by using HLA-DRA on two different breast cancer tissue microarrays (TMA): The breast cancer progression TMA provided by the Cooperative Human Tissue Network (CHTN_BrCaProg3) and the University of Illinois Breast Cancer Working Group (TMA BCWG UIC-001-TMA) that included primary tumor and lymph node metastases from patients residing in the West Side and South Side of Chicago. We also used bioinformatics analyses of the TCGA and METABRIC databases and the Broad Institute’s single-cell RNA sequencing portal to investigate MRTFA/B expression patterns in the cells of the tumor microenvironment (TME). Results We found that in human tumors, MRTFA/B are concurrently activated in cancer cells, but they show distinct patterns of expression across different histological subtypes and in the cells of the TME. Importantly, MRTFA expression was elevated in metastatic tumors of African American patients, who disproportionately die from breast cancer. Interestingly, in contrast to publicly available mRNA expression data, MRTFA was similarly expressed across estrogen receptor (ER) positive and negative breast tumors, while MRTFB expression was highest in ER+ breast tumors. Furthermore, MRTFA was specifically expressed in the perivascular antigen-presenting cells (APCs), which has been previously associated with immune suppression and breast cancer progression. We also found that MRTFA expression correlated with the expression of the immune checkpoint protein V-set immunoregulatory receptor (VSIR) in the TCGA data and found that MRTFA activity promotes VSIR expression in THP-1 monocytes and cultured HEK293 cells. Conclusions Our results provide unique insights into how MRTFA and MRTFB promote metastasis in human cancer, the differences of their expression patterns, and their immune suppressive function within the breast cancer TME. Our results will guide future studies on targeting MRTFA/B transcriptional activity and the resulting immune suppression in breast cancer. Graphical Abstracthttps://doi.org/10.1186/s12967-025-06559-3Myocardin related transcription factorsMRTFAMRTFBMALSRFMKL1
spellingShingle Stephanie M. Wilk
Kihak Lee
Caitlyn C. Castillo
Mohamed Haloul
Alexa M. Gajda
Virgilia Macias
Elizabeth L. Wiley
Zhengjia Chen
Xinyi Liu
Xiaowei Wang
Maria Sverdlov
Kent F. Hoskins
Ekrem Emrah Er
Multiplex imaging reveals novel patterns of MRTFA/B activation in the breast cancer microenvironment
Journal of Translational Medicine
Myocardin related transcription factors
MRTFA
MRTFB
MAL
SRF
MKL1
title Multiplex imaging reveals novel patterns of MRTFA/B activation in the breast cancer microenvironment
title_full Multiplex imaging reveals novel patterns of MRTFA/B activation in the breast cancer microenvironment
title_fullStr Multiplex imaging reveals novel patterns of MRTFA/B activation in the breast cancer microenvironment
title_full_unstemmed Multiplex imaging reveals novel patterns of MRTFA/B activation in the breast cancer microenvironment
title_short Multiplex imaging reveals novel patterns of MRTFA/B activation in the breast cancer microenvironment
title_sort multiplex imaging reveals novel patterns of mrtfa b activation in the breast cancer microenvironment
topic Myocardin related transcription factors
MRTFA
MRTFB
MAL
SRF
MKL1
url https://doi.org/10.1186/s12967-025-06559-3
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