PDK1 regulates cancer cell motility by antagonising inhibition of ROCK1 by RhoE

Pinner, Sophie

doi:10.1038/ncb1675

Cited by 243 publications

(238 citation statements)

References 42 publications

Supporting

Mentioning

216

Contrasting

Unclassified

Order By: Relevance

“…Using a combination of cell culture, ex vivo, and in vivo approaches, we have identified evidence supporting that Rnd3 has barrier‐protective properties under certain conditions, such as following acute inflammatory challenge. The involvement of Rnd3 in the control of endothelial barrier function adds to a growing list of recently described functions ascribed to Rnd family proteins, including Ca 2+ homeostasis in the heart,53 regulation of Notch1 signaling,54 control of cell cycle progression,55, 56, 57 development of the central nervous system,58, 59, 60 control of cell motility and metastatic potential of certain cancers,29, 61, 62, 63, 64 and a potential role in cardioprotection against heart failure 65. In addition, we provide novel evidence that delivery of Rnd3 protein can serve as a potential therapeutic tool to ameliorate microvascular hyperpermeability caused by tissue injury.…”

Section: Discussionmentioning

confidence: 99%

Rnd3 as a Novel Target to Ameliorate Microvascular Leakage

Breslin

Daines

Doggett

et al. 2016

JAHA

View full text Add to dashboard Cite

BackgroundMicrovascular leakage of plasma proteins is a hallmark of inflammation that leads to tissue dysfunction. There are no current therapeutic strategies to reduce microvascular permeability. The purpose of this study was to identify the role of Rnd3, an atypical Rho family GTPase, in the control of endothelial barrier integrity. The potential therapeutic benefit of Rnd3 protein delivery to ameliorate microvascular leakage was also investigated.Methods and ResultsUsing immunofluorescence microscopy, Rnd3 was observed primarily in cytoplasmic areas around the nuclei of human umbilical vein endothelial cells. Permeability to fluorescein isothiocyanate–albumin and transendothelial electrical resistance of human umbilical vein endothelial cell monolayers served as indices of barrier function, and RhoA, Rac1, and Cdc42 activities were determined using G‐LISA assays. Overexpression of Rnd3 significantly reduced the magnitude of thrombin‐induced barrier dysfunction, and abolished thrombin‐induced Rac1 inactivation. Depleting Rnd3 expression with siRNA significantly extended the time course of thrombin‐induced barrier dysfunction and Rac1 inactivation. Time‐lapse microscopy of human umbilical vein endothelial cells expressing GFP‐actin showed that co‐expression of mCherry‐Rnd3 attenuated thrombin‐induced reductions in local lamellipodia that accompany endothelial barrier dysfunction. Lastly, a novel Rnd3 protein delivery method reduced microvascular leakage in a rat model of hemorrhagic shock and resuscitation, assessed by both intravital microscopic observation of extravasation of fluorescein isothiocyanate–albumin from the mesenteric microcirculation, and direct determination of solute permeability in intact isolated venules.ConclusionsThe data suggest that Rnd3 can shift the balance of RhoA and Rac1 signaling in endothelial cells. In addition, our findings suggest the therapeutic, anti‐inflammatory potential of delivering Rnd3 to promote endothelial barrier recovery during inflammatory challenge.

show abstract

Section: Discussionmentioning

confidence: 99%

Rnd3 as a Novel Target to Ameliorate Microvascular Leakage

Breslin

Daines

Doggett

et al. 2016

JAHA

View full text Add to dashboard Cite

show abstract

“…For short-term lung colonization assays, melanoma cell populations were differentially labelled with 5 mM CellTracker Green CMFDA or CellTracker Orange CMTMR dyes as described 56 . Equal numbers (7.5 Â 10 5 ) were co-injected into the tail veins of nude mice, which were killed 30 min or 24 h later.…”

Section: Methodsmentioning

confidence: 99%

Tumour-derived SPARC drives vascular permeability and extravasation through endothelial VCAM1 signalling to promote metastasis

et al. 2015

View full text Add to dashboard Cite

Disruption of the endothelial barrier by tumour-derived secreted factors is a critical step in cancer cell extravasation and metastasis. Here, by comparative proteomic analysis of melanoma secretomes, we identify the matricellular protein SPARC as a novel tumour-derived vascular permeability factor. SPARC deficiency abrogates tumour-initiated permeability of lung capillaries and prevents extravasation, whereas SPARC overexpression enhances vascular leakiness, extravasation and lung metastasis. SPARC-induced paracellular permeability is dependent on the endothelial VCAM1 receptor and p38 MAPK signalling. Blocking VCAM1 impedes melanoma-induced endothelial permeability and extravasation. The clinical relevance of our findings is highlighted by high levels of SPARC detected in tumour from human pulmonary melanoma lesions. Our study establishes tumour-produced SPARC and VCAM1 as regulators of cancer extravasation, revealing a novel targetable interaction for prevention of metastasis.

show abstract

“…siRNAs against mouse Fmnl2 5 0 -AUGUGG ACGAAUUUGAAGAUU, GUGCUAAGAGAACUGGAA AUU and against human FMNL2 #1 5 0 -GUGG-UAGCAGG UAACUCUGUU and #2 GCCUGAACAACAAGAAUCC UU were from IBA. Rho GTPase SMARTpools were kindly provided by E Sahai (Cancer Research UK) and have been described earlier (Pinner and Sahai, 2008). A total of 100 nM siRNA oligos were transfected using magnet-assisted transfection (MATra) reagent according to the manufacturer's (IBA, Go¨ttingen, Germany) instructions; 48-72 h after siRNA transfection, cells were used for subsequent analysis.…”

Section: Methodsmentioning

confidence: 99%

Formin-like 2 drives amoeboid invasive cell motility downstream of RhoC

et al. 2010

View full text Add to dashboard Cite

Invasive cell migration is a key step for cancer metastasis and involves Rho GTPase-controlled reorganization of the actin cytoskeleton. Altered Rho GTPase expression is found in various malignancies. Particularly, the closely related GTPases RhoA and RhoC are upregulated in many aggressive tumours, but specific effectors that distinguish between these two GTPases to explain mechanistic differences have not been identified. The formins are by far the largest family of Rho GTPase effectors and are characterized by the actin-nucleating formin homology 2 domain. Using siRNA-based screening against all 15 human formins, we systematically analysed their functions in 3D cell motility using three different cancer cell lines. These results reveal distinct requirements for specific formins in amoeboid versus mesenchymal invasive cell migration. Importantly, by knocking down all Rho proteins, we identified formin-like 2 (FMNL2) as a specific RhoC effector, showing selective interaction of FMNL2 with active RhoC, but not RhoA or RhoB. Functional analysis shows that RhoC regulates autoinhibition of FMNL2, whereas suppression of FMNL2 inhibits RhoC-, but not RhoA-dependent, rounded invasive cell migration. Thus, our data uncover a novel regulatory and functional interaction between RhoC and FMNL2 for modulating cell shape and invasiveness and provide mechanistic insight into RhoC-specific signalling events.

show abstract

PDK1 regulates cancer cell motility by antagonising inhibition of ROCK1 by RhoE

Cited by 243 publications

References 42 publications

Rnd3 as a Novel Target to Ameliorate Microvascular Leakage

Rnd3 as a Novel Target to Ameliorate Microvascular Leakage

Tumour-derived SPARC drives vascular permeability and extravasation through endothelial VCAM1 signalling to promote metastasis

Formin-like 2 drives amoeboid invasive cell motility downstream of RhoC

Contact Info

Product

Resources

About