Vimentin intermediate filaments modulate cell traction force but not cell sensitivity to substrate stiffness

Thanh, Minh-Tri Ho; Grella, Alexandra; Kole, Denis; Ambady, Sakthikumar

doi:10.1002/cm.21675

Cited by 15 publications

(15 citation statements)

References 38 publications

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“…To estimate the forces exerted by the colony on the substrate, the stress at the surface was reconstructed from the tracer displacement maps via the finite element method (FEM) ( 47 – 49 ) (Methods). The stress at the surface increased over time, and—surprisingly—the stress was 10-fold higher for colonies on stiff substrates compared to soft ones (Fig.…”

Section: Resultsmentioning

confidence: 99%

Spreading rates of bacterial colonies depend on substrate stiffness and permeability

et al. 2022

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The ability of bacteria to colonize and grow on different surfaces is an essential process for biofilm development. Here, we report the use of synthetic hydrogels with tunable stiffness and porosity to assess physical effects of the substrate on biofilm development. Using time-lapse microscopy to track the growth of expanding Serratia marcescens colonies, we find that biofilm colony growth can increase with increasing substrate stiffness, unlike what is found on traditional agar substrates. Using traction force microscopy-based techniques, we find that biofilms exert transient stresses correlated over length scales much larger than a single bacterium, and that the magnitude of these forces also increases with increasing substrate stiffness. Our results are consistent with a model of biofilm development in which the interplay between osmotic pressure arising from the biofilm and the poroelastic response of the underlying substrate controls biofilm growth and morphology.

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Section: Resultsmentioning

confidence: 99%

Spreading rates of bacterial colonies depend on substrate stiffness and permeability

et al. 2022

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“…VIM has been implicated in two major cellular functions: mesenchymal cell motility and preservation of nuclear-cytoskeletal integrity, especially in response to compressive stress. It has also been found to play an important role in cell spreading on firm 2D substrates [62][63][64][65][66][67][68][69][70].…”

Section: Cells In the Pacc State Have Increased Vimentin Expressionmentioning

confidence: 99%

Cells in the Polyaneuploid Cancer Cell (PACC) state have increased metastatic potential

Mallin

Kim

Choudhury

et al. 2022

Preprint

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Although metastasis is the leading cause of cancer death, it is quite rare at the cellular level. Only a rare subset of cancer cells (~1 in 1.5 billion) can complete the entire metastatic cascade: invasion, intravasation, survival in the circulation, extravasation, and colonization (i.e. are metastasis competent). We propose that cells engaging a Polyaneuploid Cancer Cell (PACC) phenotype are metastasis competent. PACCs are enlarged, non-dividing cells with increased genomic content that form in response to stress. Single-cell tracking using time-lapse microscopy reveals that PACCs are more motile than nonPACCs. Additionally, PACCs exhibit increased capacity for environment-sensing and directional migration in chemotactic environments, predicting successful invasion. Magnetic Twisting Cytometry and Atomic Force Microscopy reveal that cells in the PACC state display hyper-elastic properties like increased peripheral deformability and maintained peri-nuclear cortical integrity that predict successful intravasation and extravasation. Furthermore, four orthogonal methods reveal that PACCs have increased expression of Vimentin, a known hyper-elastic biomolecule. Lastly, anoikis-resistance assays and detection of PACCs in the blood of a patient with metastatic castrate-resistant prostate cancer using a selection-free circulating tumor cell detection platform reveal that PACCs are capable of surviving in the circulation. Taken together with the knowledge that PACCs are capable of eventual depolyploidization and progeny formation (as a potential route to colonization), these data support PACCs as candidate metastasis-competent cells worthy of further analysis.

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“…On the other hand, PLN and MYH6 CM-specific gene expression was not detected in NKX2.5 GFP -cFib when compared with CBiPS5 NKX2.5 GFP -CM (Figure 3E). In contrast, NKX2.5 GFP -cFib expressed fibroblast-specific markers such as THY1 (cell surface marker for distinct fibroblast populations), POSTN (expressed in cFib during embryogenesis (Snider et al, 2009) and increeased in adult cFib upon cardiac injury (Kanisicak et al, 2016;Fu et al, 2018)), VIM (an intermediate filament that modulates fibroblast traction force (Ho Thanh et al, 2021)) and TCF21 (a transcription factor required for cFib development (Acharya et al, 2012;Kanisicak et al, 2016)). We investigated the expression of these fibroblast-specific markers in NKX2.5 GFP -cFib during the first three passages.…”

Section: Generation Of Functional Nkx25 Gfp Reporter Human Fibroblastsmentioning

confidence: 99%

Generation of NKX2.5GFP Reporter Human iPSCs and Differentiation Into Functional Cardiac Fibroblasts

López-Muneta

Linares

Casis

et al. 2022

Front. Cell Dev. Biol.

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Direct cardiac reprogramming has emerged as an interesting approach for the treatment and regeneration of damaged hearts through the direct conversion of fibroblasts into cardiomyocytes or cardiovascular progenitors. However, in studies with human cells, the lack of reporter fibroblasts has hindered the screening of factors and consequently, the development of robust direct cardiac reprogramming protocols.In this study, we have generated functional human NKX2.5GFP reporter cardiac fibroblasts. We first established a new NKX2.5GFP reporter human induced pluripotent stem cell (hiPSC) line using a CRISPR-Cas9-based knock-in approach in order to preserve function which could alter the biology of the cells. The reporter was found to faithfully track NKX2.5 expressing cells in differentiated NKX2.5GFP hiPSC and the potential of NKX2.5-GFP + cells to give rise to the expected cardiac lineages, including functional ventricular- and atrial-like cardiomyocytes, was demonstrated. Then NKX2.5GFP cardiac fibroblasts were obtained through directed differentiation, and these showed typical fibroblast-like morphology, a specific marker expression profile and, more importantly, functionality similar to patient-derived cardiac fibroblasts. The advantage of using this approach is that it offers an unlimited supply of cellular models for research in cardiac reprogramming, and since NKX2.5 is expressed not only in cardiomyocytes but also in cardiovascular precursors, the detection of both induced cell types would be possible. These reporter lines will be useful tools for human direct cardiac reprogramming research and progress in this field.

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Vimentin intermediate filaments modulate cell traction force but not cell sensitivity to substrate stiffness

Cited by 15 publications

References 38 publications

Spreading rates of bacterial colonies depend on substrate stiffness and permeability

Spreading rates of bacterial colonies depend on substrate stiffness and permeability

Cells in the Polyaneuploid Cancer Cell (PACC) state have increased metastatic potential

Generation of NKX2.5GFP Reporter Human iPSCs and Differentiation Into Functional Cardiac Fibroblasts

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