TY - JOUR
T1 - Expression of vimentin alters cell mechanics, cell-cell adhesion, and gene expression profiles suggesting the induction of a hybrid EMT in human mammary epithelial cells
AU - Sivagurunathan, Suganya
AU - Vahabikashi, Amir
AU - Yang, Haiqian
AU - Zhang, Jun
AU - Vazquez, Kelly
AU - Rajasundaram, Dhivyaa
AU - Politanska, Yuliya
AU - Abdala-Valencia, Hiam
AU - Notbohm, Jacob
AU - Guo, Ming
AU - Adam, Stephen A
AU - Goldman, Robert David
N1 - Funding Information:
This work was supported by NIH P01GM096971 awarded to RG and 1R01 GM140108 awarded to MG, Ian Wong (Brown University) and RG. Imaging work was performed at the Northwestern University Center for Advanced Microscopy generously supported by CCSG P30 CA060553 awarded to the Robert H Lurie Comprehensive Cancer Center. Structured illumination microscopy was performed on a Nikon N-SIM system, purchased through the support of NIH 1S10OD016342-01. Flow cytometry work was supported by the Northwestern University RHLCCC Flow Cytometry Facility and a Cancer Center Support Grant (NCI CA060553). Flow Cytometry Cell Sorting was performed on a BD FACSAria SORP system and BD FACSymphony S6 SORP system, purchased through the support of NIH 1S10OD011996-01 and 1S10OD026814-01. RNA sequencing was carried out in the Metabolomics Core -“Integrative Genomics” at the Robert H. Lurie Comprehensive Cancer Center.
Funding Information:
This work was supported by NIH P01GM096971 awarded to RG and 1R01 GM140108 awarded to MG, Ian Wong (Brown University) and RG. Imaging work was performed at the Northwestern University Center for Advanced Microscopy generously supported by CCSG P30 CA060553 awarded to the Robert H Lurie Comprehensive Cancer Center. Structured illumination microscopy was performed on a Nikon N-SIM system, purchased through the support of NIH 1S10OD016342-01. Flow cytometry work was supported by the Northwestern University RHLCCC Flow Cytometry Facility and a Cancer Center Support Grant (NCI CA060553). Flow Cytometry Cell Sorting was performed on a BD FACSAria SORP system and BD FACSymphony S6 SORP system, purchased through the support of NIH 1S10OD011996-01 and 1S10OD026814-01. RNA sequencing was carried out in the Metabolomics Core -“Integrative Genomics” at the Robert H. Lurie Comprehensive Cancer Center.
Publisher Copyright:
Copyright © 2022 Sivagurunathan, Vahabikashi, Yang, Zhang, Vazquez, Rajasundaram, Politanska, Abdala-Valencia, Notbohm, Guo, Adam and Goldman.
PY - 2022/9/19
Y1 - 2022/9/19
N2 - Vimentin is a Type III intermediate filament (VIF) cytoskeletal protein that regulates the mechanical and migratory behavior of cells. Its expression is considered to be a marker for the epithelial to mesenchymal transition (EMT) that takes place in tumor metastasis. However, the molecular mechanisms regulated by the expression of vimentin in the EMT remain largely unexplored. We created MCF7 epithelial cell lines expressing vimentin from a cumate-inducible promoter to address this question. When vimentin expression was induced in these cells, extensive cytoplasmic VIF networks were assembled accompanied by changes in the organization of the endogenous keratin intermediate filament networks and disruption of desmosomes. Significant reductions in intercellular forces by the cells expressing VIFs were measured by quantitative monolayer traction force and stress microscopy. In contrast, laser trapping micro-rheology revealed that the cytoplasm of MCF7 cells expressing VIFs was stiffer than the uninduced cells. Vimentin expression activated transcription of genes involved in pathways responsible for cell migration and locomotion. Importantly, the EMT related transcription factor TWIST1 was upregulated only in wild type vimentin expressing cells and not in cells expressing a mutant non-polymerized form of vimentin, which only formed unit length filaments (ULF). Taken together, our results suggest that vimentin expression induces a hybrid EMT correlated with the upregulation of genes involved in cell migration.
AB - Vimentin is a Type III intermediate filament (VIF) cytoskeletal protein that regulates the mechanical and migratory behavior of cells. Its expression is considered to be a marker for the epithelial to mesenchymal transition (EMT) that takes place in tumor metastasis. However, the molecular mechanisms regulated by the expression of vimentin in the EMT remain largely unexplored. We created MCF7 epithelial cell lines expressing vimentin from a cumate-inducible promoter to address this question. When vimentin expression was induced in these cells, extensive cytoplasmic VIF networks were assembled accompanied by changes in the organization of the endogenous keratin intermediate filament networks and disruption of desmosomes. Significant reductions in intercellular forces by the cells expressing VIFs were measured by quantitative monolayer traction force and stress microscopy. In contrast, laser trapping micro-rheology revealed that the cytoplasm of MCF7 cells expressing VIFs was stiffer than the uninduced cells. Vimentin expression activated transcription of genes involved in pathways responsible for cell migration and locomotion. Importantly, the EMT related transcription factor TWIST1 was upregulated only in wild type vimentin expressing cells and not in cells expressing a mutant non-polymerized form of vimentin, which only formed unit length filaments (ULF). Taken together, our results suggest that vimentin expression induces a hybrid EMT correlated with the upregulation of genes involved in cell migration.
KW - cell-cell adhesion
KW - desmoplakin
KW - hybrid/partial EMT
KW - intercellular forces
KW - intracellular mechanics
KW - Twist1
KW - vimentin
UR - http://www.scopus.com/inward/record.url?scp=85139212054&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85139212054&partnerID=8YFLogxK
U2 - 10.3389/fcell.2022.929495
DO - 10.3389/fcell.2022.929495
M3 - Article
C2 - 36200046
AN - SCOPUS:85139212054
VL - 10
JO - Frontiers in Cell and Developmental Biology
JF - Frontiers in Cell and Developmental Biology
SN - 2296-634X
M1 - 929495
ER -