c-Abl, Lamellipodin, and Ena/VASP Proteins Cooperate in Dorsal Ruffling of Fibroblasts and Axonal Morphogenesis

Michael, Magdalene; Vehlow, Anne; Navarro, Christel; Krause, Matthias

doi:10.1016/j.cub.2010.03.048

Cited by 74 publications

(109 citation statements)

References 36 publications

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“…Nonetheless, based on the varying endothelial barrier properties and adhesion molecules used for extravasation into different tissues, it is possible that the Ena/ VASP family may have a more or less profound impact on trafficking to different anatomical sites. The observation that deletion of EVL and VASP impaired activated T-cell trafficking in vivo is consistent with the literature implicating Ena/VASP proteins in cellular motility (27,(47)(48)(49)(50)(51). Although we identified reduced actin polymerization in response to chemokine triggering in activated EVL/VASP dKO T cells, surprisingly we found no major effect on activated T-cell chemotaxis or crawling in vitro.…”

Section: Discussionsupporting

confidence: 92%

Ena/VASP proteins regulate activated T-cell trafficking by promoting diapedesis during transendothelial migration

Estin

Thompson

Traxinger

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Vasodilator-stimulated phosphoprotein (VASP) and Ena-VASP-like (EVL) are cytoskeletal effector proteins implicated in regulating cell morphology, adhesion, and migration in various cell types. However, the role of these proteins in T-cell motility, adhesion, and in vivo trafficking remains poorly understood. This study identifies a specific role for EVL and VASP in T-cell diapedesis and trafficking. We demonstrate that EVL and VASP are selectively required for activated T-cell trafficking but are not required for normal T-cell development or for naïve T-cell trafficking to lymph nodes and spleen. Using a model of multiple sclerosis, we show an impairment in trafficking of EVL/VASP-deficient activated T cells to the inflamed central nervous system of mice with experimental autoimmune encephalomyelitis. Additionally, we found a defect in trafficking of EVL/VASP double-knockout (dKO) T cells to the inflamed skin and secondary lymphoid organs. Deletion of EVL and VASP resulted in the impairment in α4 integrin (CD49d) expression and function. Unexpectedly, EVL/VASP dKO T cells did not exhibit alterations in shear-resistant adhesion to, or in crawling on, primary endothelial cells under physiologic shear forces. Instead, deletion of EVL and VASP impaired T-cell diapedesis. Furthermore, T-cell diapedesis became equivalent between control and EVL/VASP dKO T cells upon α4 integrin blockade. Overall, EVL and VASP selectively mediate activated T-cell trafficking by promoting the diapedesis step of transendothelial migration in a α4 integrin-dependent manner.A ctivated T-cell trafficking across the vascular endothelium is essential for ongoing immune surveillance of tissues and for effective immune responses to conditions such as infection and cancer. Conversely, in situations of immune dysregulation, inhibition of self-reactive T-cell trafficking represents a promising target for therapeutic immunomodulation. Disruption of these pathways, such as by antibody blockade of α4 integrins, is a highly effective approach to immunomodulation (1, 2). However, the molecular mechanisms by which chemokine receptor and adhesion molecule signaling induce the T-cell cytoskeletal machinery to promote extravasation are not yet fully elucidated.Transendothelial migration (TEM), the process by which T cells extravasate from the blood into tissues, is characterized by four distinct steps: rolling along the vascular wall, arrest or adhesion, intravascular crawling, and diapedesis across the endothelial barrier (3). Surface adhesion molecules play well-characterized roles in each step of the process. For example, the initial rolling step of TEM is facilitated by interactions between T-cell and endothelial selectins, whereas the adhesion, intravascular crawling, and diapedesis steps of TEM are mainly regulated by chemokineand shear force-stimulated modulation of lymphocyte functionassociated antigen 1 (LFA-1, αLβ2 integrin, CD11a/CD18) and very late antigen 4 (VLA-4, α4β1 integrin, CD49d/CD29) interactions with intracellular adhesion molecule 1 ...

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Section: Discussionsupporting

confidence: 92%

Ena/VASP proteins regulate activated T-cell trafficking by promoting diapedesis during transendothelial migration

Estin

Thompson

Traxinger

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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“…Lamellipodin, a phosphoinositide binding protein, was previously shown to play an important role in recruiting Ena/VASP to the leading edge in fibroblasts and melanoma cells (note that lamellipodin targets to the leading edge independently of Ena/ VASP) (21,22). Lamellipodin depletion by siRNA markedly reduced VASP accumulation at the leading edge in profilin1 knockdown MDA-231 cells (Fig.…”

Section: Profilin1 Inhibits Mda-mb-231 Cell Motility By Attenuatingmentioning

confidence: 80%

“…Activation of receptor tyrosine kinases (RTKs) such as EGFR (EGF receptor) and PDGFR (PDGF receptor) lead to PI3 kinase (PI3K)-mediated generation of D3-phosphoinositides, including PI(3,4,5)P 3 and PI(3,4)P 2 [the latter mostly through PI(3,4,5)P 3 dephosphorylation by 5′-phosphatases]. Consistent with this, PDGF stimulation has been shown to recruit lamellipodin or even the isolated PH domain of lamellipodin to the leading edge and the tips of dorsal ruffles (21,22). We found that PI3K inhibition by LY294002 dramatically inhibits lamellipodin accumulation at the leading edge in profilin1 knockdown MDA-231 cells (with or without rescue by R88L-profilin1) under PDGF-stimulated condition (Fig.…”

Section: Profilin1 Inhibits Mda-mb-231 Cell Motility By Attenuatingmentioning

confidence: 83%

Profilin1 regulates PI(3,4)P ₂ and lamellipodin accumulation at the leading edge thus influencing motility of MDA-MB-231 cells

Bae

Ding

Das

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

109

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Profilin1, a ubiquitously expressed actin-binding protein, plays a critical role in cell migration through actin cytoskeletal regulation. Given the traditional view of profilin1 as a promigratory molecule, it is difficult to reconcile observations that profilin1 is down-regulated in various invasive adenocarcinomas and that reduced profilin1 expression actually confers increased motility to certain adenocarcinoma cells. In this study, we show that profilin1 negatively regulates lamellipodin targeting to the leading edge in MDA-MB-231 breast cancer cells and normal cells; profilin1 depletion increases lamellipodin concentration at the lamellipodial tip (where it binds Ena/VASP), and this mediates the hypermotility. We report that the molecular mechanism underlying profilin1's modulation of lamellipodin localization relates to phosphoinositide control. Specifically, we show that phosphoinositide binding of profilin1 inhibits the motility of MDA-MB-231 cells by negatively regulating PI(3,4)P 2 at the membrane and thereby limiting recruitment of lamellipodin [a PI(3,4)P 2 -binding protein] and Ena/ VASP to the leading edge. In summary, this study uncovers a unique biological consequence of profilin1-phosphoinositide interaction, thus providing direct evidence of profilin1's regulation of cell migration independent of its actin-related activity.T he ubiquitously expressed cytoskeleton-modulating protein profilin1 influences multiple processes involved in cell motility, making it a challenge to elucidate the exact molecular mechanism that controls migration. At least one major function of profilin1 is to regulate actin polymerization. Profilin1 regenerates actin monomers from disassembling filament networks by facilitating the exchange of ATP for ADP on G-actin. By further inhibiting spontaneous nucleation of G-actin, profilin1 causes an accumulation of profilin1/ATP-G-actin pool available for polymerization. Because profilin1 also has an affinity for poly-Lproline sequences, it binds to almost all major actin nucleating and F-actin elongating proteins that contain proline-rich domains [e.g., N-WASP (neuronal Wiskott-Aldrich syndrome protein), Ena (enabled)/VASP (vasodilator stimulated phosphoprotein), and formins], and this allows profilin1-mediated recruitment of ATP-G-actin to these proteins, enhancing actin polymerization (1, 2). In addition, profilin1 binds to plasma membrane presumably through its interactions with various phosphoinositides (3). Profilin1 binds to phosphatidylinositol-4,5-bisphosphate [PI(4,5)P 2 ], phosphatidylinositol-3,4-bisphosphate [PI(3,4)P 2 ], and phosphatidylinositol-3,4,5-triphosphate [PI(3,4,5) P 3 ]), at least in vitro (4). Based on PI(4,5)P 2 binding, it has been proposed that the phosphoinositide binding site of profilin1 overlaps with its actin-binding site (5), and to some extent spans a second region neighboring the polyproline binding site (6). This has prompted speculation that the major role of phosphoinositide binding of profilin1 would be to inhibit its interaction with act...

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“…In primary cortical neurons, Lpd promotes axonogenesis, but the relationship with PI(3,4)P 2 has not been examined (Michael et al, 2010). Thus, the direct role of PI(3,4)P 2 in the process formation of vertebrate neurons has yet to be explored.…”

Section: Discussionmentioning

confidence: 99%

“…Ena/vasodilator-stimulated phosphoprotein (VASP) proteins antagonize capping proteins at actin filament-barbed ends and regulate filopodia and lamellipodia (Michael et al, 2010). Lpd is located in the phosphatidylinositol (3,4)-bisphosphate [PI(3,4)P 2 ]-rich region on the plasma membrane and recruits Ena/VASP proteins .…”

Section: Introductionmentioning

confidence: 99%

A Phosphatidylinositol Lipids System, Lamellipodin, and Ena/VASP Regulate Dynamic Morphology of Multipolar Migrating Cells in the Developing Cerebral Cortex

et al. 2012

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In the developing mammalian cerebral cortex, excitatory neurons are generated in the ventricular zone (VZ) and subventricular zone; these neurons migrate toward the pial surface. The neurons generated in the VZ assume a multipolar morphology and remain in a narrow region called the multipolar cell accumulation zone (MAZ) for ϳ24 h, in which they extend and retract multiple processes dynamically. They eventually extend an axon tangentially and begin radial migration using a migratory mode called locomotion. Despite the potential biological importance of the process movement of multipolar cells, the molecular mechanisms remain to be elucidated. Here, we observed that the processes of mouse multipolar cells were actin rich and morphologically resembled the filopodia and lamellipodia in growth cones; thus, we focused on the actin-remodeling proteins Lamellipodin (Lpd) and Ena/vasodilator-stimulated phosphoprotein (VASP). Lpd binds to phosphatidylinositol (3,4)-bisphosphate [PI(3,4)P 2 ] and recruits Ena/VASP, which promotes the assembly of actin filaments, to the plasma membranes. In situ hybridization and immunohistochemistry revealed that Lpd is expressed in multipolar cells in the MAZ. The functional silencing of either Lpd or Ena/VASP decreased the number of primary processes. Immunostaining and a Förster resonance energy transfer analysis revealed the subcellular localization of PI(3,4)P 2 at the tips of the processes. A knockdown experiment and treatment with an inhibitor for Src homology 2-containing inositol phosphatase-2, a 5-phosphatase that produces PI(3,4)P 2 from phosphatidylinositol (3,4,5)-triphosphate, decreased the number of primary processes. Our observations suggest that PI(3,4)P 2 , Lpd, and Ena/VASP are involved in the process movement of multipolar migrating cells.

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c-Abl, Lamellipodin, and Ena/VASP Proteins Cooperate in Dorsal Ruffling of Fibroblasts and Axonal Morphogenesis

Cited by 74 publications

References 36 publications

Ena/VASP proteins regulate activated T-cell trafficking by promoting diapedesis during transendothelial migration

Ena/VASP proteins regulate activated T-cell trafficking by promoting diapedesis during transendothelial migration

Profilin1 regulates PI(3,4)P ₂ and lamellipodin accumulation at the leading edge thus influencing motility of MDA-MB-231 cells

A Phosphatidylinositol Lipids System, Lamellipodin, and Ena/VASP Regulate Dynamic Morphology of Multipolar Migrating Cells in the Developing Cerebral Cortex

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c-Abl, Lamellipodin, and Ena/VASP Proteins Cooperate in Dorsal Ruffling of Fibroblasts and Axonal Morphogenesis

Cited by 74 publications

References 36 publications

Ena/VASP proteins regulate activated T-cell trafficking by promoting diapedesis during transendothelial migration

Ena/VASP proteins regulate activated T-cell trafficking by promoting diapedesis during transendothelial migration

Profilin1 regulates PI(3,4)P 2 and lamellipodin accumulation at the leading edge thus influencing motility of MDA-MB-231 cells

A Phosphatidylinositol Lipids System, Lamellipodin, and Ena/VASP Regulate Dynamic Morphology of Multipolar Migrating Cells in the Developing Cerebral Cortex

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Profilin1 regulates PI(3,4)P ₂ and lamellipodin accumulation at the leading edge thus influencing motility of MDA-MB-231 cells