Hydraulic resistance induces cell phenotypic transition in confinement

Abstract

Cells penetrating into confinement undergo mesenchymal-to-amoeboid transition. The topographical features of the microenvironment expose cells to different hydraulic resistance levels. How cells respond to hydraulic resistance is unknown. We show that the cell phenotype shifts from amoeboid to mesenchymal upon increasing resistance. By combining automated morphological tracking and wavelet analysis along with fluorescence recovery after photobleaching (FRAP), we found an oscillatory phenotypic transition that cycles from blebbing to short, medium, and long actin network formation, and back to blebbing. Elevated hydraulic resistance promotes focal adhesion maturation and long actin filaments, thereby reducing the period required for amoeboid-to-mesenchymal transition. The period becomes independent of resistance upon blocking the mechanosensor TRPM7. Mathematical modeling links intracellular calcium oscillations with actomyosin turnover and force generation and recapitulates experimental data. We identify hydraulic resistance as a critical physical cue controlling cell phenotype and present an approach for connecting fluorescent signal fluctuations to morphological oscillations.

The fluid shear stress sensor TRPM7 regulates tumor cell intravasation

“…These authors reported that TRPM7 knockdown conferred a "contractile" phenotype, suppressed migration velocity and wound closure, and metastasis (45). These findings are in distinct contrast to (i) the "slender" shape observed in prior studies (8,40) and in this work, (ii) our previous data showing that TRPM7 promotes actomyosin contractility (9), and (iii) the slower track velocity and displacement at the front of in vivo tumors of HT-1080 cells overexpressing TRPM7. It is also noteworthy that Kaplan-Meier analysis of human patients shows that patients with osteosarcoma, breast, gastric, and liver cancer expressing high levels of TRPM7 had improved survival (fig.…”

“…In line with these observations, chelation of extracellular calcium with BAPTA or inhibition of ROCK activity or myosin-II function blocked the shear-induced directional change of fibroblasts, thereby promoting their intravasation in the presence of shear flow. Although fibroblasts typically exhibit a mesenchymal phenotype, their exposure to fluid shear triggers RhoA activation, which supports a bleb-based phenotype (17,18,40). Along these lines, half of the fibroblasts, which reversed migration direction to avert exposure to fluid flow, displayed a blebbing phenotype.…”

Effective Force Generation During Mammalian Cell Migration Under Different Molecular and Physical Mechanisms

“…This difference suggests that cells will reorganize their cytoskeleton structure upon external mechanical input. Experimentally, it has indeed been observed that the actin network can further polarize towards the front of the cell when cells experience high external hydrostatic pressure ( Zhao et al, 2019 ; Zhao et al, 2021 ).…”

The interplay between physical cues and mechanosensitive ion channels in cancer metastasis

“…Increased cation entry can affect cellular function directly or by enhancing intracellular signaling cascades which lead to the downstream release of calcium ( Pedersen and Nilius, 2007 ; Petho et al, 2019 ). Importantly, TRP channels also regulate the proliferation, differentiation, migration, invasion and chemoresistance of cancer cells by sensing osmotic perturbations, shear forces and hydrostatic pressure ( Clapham, 2003 ; Liedtke et al, 2003 ; Gomis et al, 2008 ; Mendoza et al, 2010 ; Shen et al, 2015 ; Zhao et al, 2019 ; Yankaskas et al, 2021 ; Zhao et al, 2021 ) ( Table 1 ). Several members of the TRP superfamily also participate in actin remodeling and focal adhesion dynamics in response to mechanical stimuli ( Kobayashi and Sokabe, 2010 ; Shen et al, 2015 ; Zhao et al, 2019 ).…”

“…CRISPR/Cas9 knockout of TRPM7 blinds cells to hydraulic resistance and causes them to choose migratory paths based on cross-sectional area ( Zhao et al, 2019 ). TRPM7 not only influences the directional navigation of cancer cells but also provides cells with plasticity to counter elevated hydraulic resistances ( Zhao et al, 2021 ). Elevated hydraulic resistance induces a shift in cell migration phenotype from amoeboid to mesenchymal ( Zhao et al, 2021 ), which relies on the formation of actin-rich lamellipodia and integrin-based cell-matrix adhesions rather than membrane blebs ( Friedl and Wolf, 2010 ; Pankova et al, 2010 ).…”