Feedback-Driven Mechanisms Between Phosphorylated Caveolin-1 and Contractile Actin Assemblies Instruct Persistent Cell Migration

Zhao

et al. 2021

Front. Cell Dev. Biol.

Mesenchymal stem cells (MSCs) secrete cytokines in a paracrine or autocrine manner to regulate immune response and tissue regeneration. Our previous research revealed that MSCs use the complex of Fas/Fas-associated phosphatase-1 (Fap-1)/caveolin-1 (Cav-1) mediated exocytotic process to regulate cytokine and small extracellular vesicles (EVs) secretion, which contributes to accelerated wound healing. However, the detailed underlying mechanism of cytokine secretion controlled by Cav-1 remains to be explored. We show that Gingiva-derived MSCs (GMSCs) could secrete more C-X-C motif chemokine ligand 10 (CXCL10) but showed lower phospho-Cav-1 (p-Cav-1) expression than skin-derived MSCs (SMSCs). Moreover, dephosphorylation of Cav-1 by a Src kinase inhibitor PP2 significantly enhances CXCL10 secretion, while activating phosphorylation of Cav-1 by H2O2 restraints CXCL10 secretion in GMSCs. We also found that Fas and Fap-1 contribute to the dephosphorylation of Cav-1 to elevate CXCL10 secretion. Tumor necrosis factor-α serves as an activator to up-regulate Fas, Fap-1, and down-regulate p-Cav-1 expression to promote CXCL10 release. Furthermore, local applying p-Cav-1 inhibitor PP2 could accelerate wound healing, reduce the expression of α-smooth muscle actin and increase cleaved-caspase 3 expression. These results indicated that dephosphorylation of Cav-1 could inhibit fibrosis during wound healing. The present study establishes a previously unknown role of p-Cav-1 in controlling cytokine release of MSC and may present a potential therapeutic approach for promoting scarless wound healing.

Section: Resultsmentioning

confidence: 99%

Dephosphorylation of Caveolin-1 Controls C-X-C Motif Chemokine Ligand 10 Secretion in Mesenchymal Stem Cells to Regulate the Process of Wound Healing

Zhao

et al. 2021

Front. Cell Dev. Biol.

“…Next, we explored the mechanism by which the NS1643-induced dephosphorylation of Cav-1 inhibits cancer cell migration. Although the phosphorylation of Cav-1 was shown to promote cell migration by facilitating actin remodeling, focal adhesion turnover, pseudopodial protrusions, and stress fiber formation [ 25 , 48 ], in a previous study, we showed that the activation of Kv11.1 significantly increases β-catenin levels at the junctional membrane in MCF-7 cells [ 9 ]. Here, we observed the same effect of NS1643 in triple negative MDA-MB-231 cells, which was absent in Cav-1 depleted cells.…”

Section: Discussionmentioning

confidence: 99%

“…Phosphorylated Cav-1 reciprocally enhances RhoA activity and facilitates focal adhesion turnover, leading to enhanced tumor cell migration. Cytoplasmic Cav-1 was shown to be associated with contractile actin filaments [ 25 ]. These authors showed that cytoplasmic Cav-1-positive vesicles move along actin filaments, but once Cav-1 is dephosphorylated, the motility of these vesicles is decreased, due to reduced RhoA-myosin Ⅱ activity and increased Rac1-PAK1-Cofillin activation, resulting in the disorganization of contractile stress fibers and compromised directional movement.…”

Section: Introductionmentioning

confidence: 99%

Pharmacological Activation of Potassium Channel Kv11.1 with NS1643 Attenuates Triple Negative Breast Cancer Cell Migration by Promoting the Dephosphorylation of Caveolin-1

Jiang

Senyuk

et al. 2022

Cells

The prevention of metastasis is a central goal of cancer therapy. Caveolin-1 (Cav-1) is a structural membrane and scaffolding protein shown to be a key regulator of late-stage breast cancer metastasis. However, therapeutic strategies targeting Cav-1 are still lacking. Here, we demonstrate that the pharmacological activation of potassium channel Kv11.1, which is uniquely expressed in MDA-MB-231 triple negative breast cancer cells (TNBCs) but not in normal MCF-10A cells, induces the dephosphorylation of Cav-1 Tyr-14 by promoting the Ca2+-dependent stimulation of protein tyrosine phosphatase 1B (PTP1B). Consequently, the dephosphorylation of Cav-1 resulted in its disassociation from β-catenin, which enabled the accumulation of β-catenin at cell borders, where it facilitated the formation of cell–cell adhesion complexes via interactions with R-cadherin and desmosomal proteins. Kv11.1 activation-dependent Cav-1 dephosphorylation induced with NS1643 also reduced cell migration and invasion, consistent with its ability to regulate focal adhesion dynamics. Thus, this study sheds light on a novel pharmacological mechanism of promoting Cav-1 dephosphorylation, which may prove to be effective at reducing metastasis and promoting contact inhibition.

Journal of Molecular Cell Biology

“…Previous studies revealed the intimate links between caveolae/CAV-1 and stress fibers ( Echarri and Del Pozo, 2015 ). Our previous work also found that CAV-1 knockout led to disordered stress fibers and prominent lamellipodia by regulating the active level of RhoA–myosin II and Rac1–PAK1–Cofilin ( Shi et al, 2021 ). Inhibition of stress fibers, but not Arp2/3-dependent branched actin filaments, diminished the phosphorylation of CAV-1 on site Tyr14 ( Shi et al, 2021 ).…”

Section: Introductionmentioning

confidence: 86%

“…Our previous work also found that CAV-1 knockout led to disordered stress fibers and prominent lamellipodia by regulating the active level of RhoA–myosin II and Rac1–PAK1–Cofilin ( Shi et al, 2021 ). Inhibition of stress fibers, but not Arp2/3-dependent branched actin filaments, diminished the phosphorylation of CAV-1 on site Tyr14 ( Shi et al, 2021 ). It has been reported that CAV-1-regulated actin-related mechanosensitive pathways was closely related to its regulation on RhoA activity ( Peng et al, 2007 ).…”

Section: Introductionmentioning

confidence: 86%

Actin nucleator formins regulate the tension-buffering function of caveolin-1

Shi

Tang

Xing

et al. 2021

Self Cite

Both the mechanosensitive actin cytoskeleton and caveolae contribute to active processes such as cell migration, morphogenesis, and vesicular trafﬁcking. Although distinct actin components are well studied, how they contribute to cytoplasmic caveolae, especially in the context of mechano-stress, has remained elusive. Here, we identify two actin-associated mobility stereotypes of caveolin-1 (CAV-1)-marked intracellular vesicles, which are characterized as ‘dwelling’ and ‘go and dwelling’. In order to exploit the reason for their distinct dynamics, elongated actin-associated formin functions are perturbed. We find drastically decreased density, increased clustering, and compromised motility of cytoplasmic CAV-1 vesicles resulting from lacking actin nucleator formins by both chemical treatment and RNA silencing of formin genes. Furthermore, hypo-osmosis-stimulated diminishing of CAV-1 is dramatically intensified upon blocking formins. The clustering of CAV-1 vesicles when cells are cultured on soft substrate is also aggravated under formin inhibition condition. Together, we reveal that actin-associated formins are essential for maintaining the dynamic organization of cytoplasmic CAV-1 and importantly its sensitivity upon mechanical challenge. We conclude that tension-controlled actin formins act as a safety valve dampening excessive tension on CAV-1 and safeguarding CAV-1 against mechanical damage.