Polarized migration of lymphatic endothelial cells is critically dependent on podoplanin regulation of Cdc42

Navarro, Angels; Perez, Ricardo E.; Rezaiekhaligh, Mohammad H.; Mabry, Sherry M; Ekekezie, Ikechukwu I

doi:10.1152/ajplung.00171.2010

Cited by 37 publications

(42 citation statements)

References 39 publications

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“…**=p < 0.01 compared to no treatment by Dunnett´s test. level in other reports judging by the unquantified Western blots presented [17,18]. Nevertheless, this reduction in expression was sufficient to negate the effect of VEGF on migration.…”

Section: Discussionmentioning

confidence: 58%

“…In our studies, crosslinking of podoplanin on LEC (where there would be no substances from platelets) was similarly effective to platelets themselves, ruling out an absolute requirement for released agents in effects described. Others noted that knockdown of podoplanin in LEC reduced their ability to migrate across a wound or form networks on Matrigel [17,18]. In MDCK cells, overexpression of podoplanin increased active RhoA and phosphorylated ERM proteins [15], while in LEC, podoplanin knockdown was associated with a reduction in active RhoA but had no effect on the amount of phosphorylated ERM proteins [16] We did not find any change in phosphorylated ERM proteins after podoplanin knockdown, while crosslinking podoplanin did not inhibit the increase in active RhoA detected within 10 minutes or 10 hours after treatment with VEGFC (data not shown).…”

Section: Discussionmentioning

confidence: 99%

“…Overexpression of podoplanin in Madin-Darby canine kidney (MDCK) cells promoted migration and increased the amount of active RhoA, which was associated with an increase in phosphorylation of ERM proteins, leading the authors to conclude that podoplanin phosphorylates ERM proteins via RhoA and its effector protein, Rho kinase (ROCK) [15]. Conversely, it has been shown that siRNA-mediated knockdown of podoplanin in LEC reduced the ability of these cells to migrate across a wound or form networks on Matrigel [17,18]. Podoplanin knockdown was associated with a reduction in active RhoA and treating LEC with an inhibitor of RhoA was able to prevent network formation [17].…”

Section: Introductionmentioning

confidence: 99%

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Modulation of VEGF-induced migration and network formation by lymphatic endothelial cells: Roles of platelets and podoplanin

et al. 2017

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Lymphatic endothelial cells (LEC) express the transmembrane receptor podoplanin whose only known endogenous ligand CLEC-2 is found on platelets. Both podoplanin and CLEC-2 are required for normal lymphangiogenesis as mice lacking either protein develop a blood-lymphatic mixing phenotype. We investigated the roles of podoplanin and its interaction with platelets in migration and tube formation by LEC. Addition of platelets or antibody-mediated crosslinking of podoplanin inhibited LEC migration induced by vascular endothelial growth factors (VEGF-A or VEGF-C), but did not modify basal migration or the response to basic fibroblast growth factor or epidermal growth factor. In addition, platelets and podoplanin crosslinking disrupted networks of LEC formed in co-culture with fibroblasts. Depletion of podoplanin in LEC using siRNA negated the pro-migratory effect of VEGF-A and VEGF-C. Inhibition of RhoA or Rho-kinase reduced LEC migration induced by VEGF-C, but had no further effect after crosslinking of podoplanin, suggesting that podoplanin is required for signaling downstream of VEGF-receptors but upstream of RhoA. Together, these data reveal for the first time that podoplanin is an intrinsic specific regulator of VEGF-mediated migration and network formation in LEC and identify crosslinking of podoplanin by platelets or antibodies as mechanisms to modulate this pathway.

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Section: Discussionmentioning

confidence: 58%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Modulation of VEGF-induced migration and network formation by lymphatic endothelial cells: Roles of platelets and podoplanin

et al. 2017

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“…Podoplanin up-regulation in cancer cells is associated with altered actin cytoskeleton reorganization and increased tumor cell migration and invasiveness. 46 Similarly, in lung microvascular LECs, small interfering RNA-mediated podoplanin knockdown causes a dramatic reduction in directional migration 47 and abrogated formation of capillary tubes on Matrigel. 48 These effects appear to be independent of ligand engagement and may reflect constitutive signaling from podoplanin.…”

Section: Discussionmentioning

confidence: 99%

CLEC-2 and Syk in the megakaryocytic/platelet lineage are essential for development

Finney

Schweighoffer

Navarro-Núñez

et al. 2012

Blood

134

139

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The C-type lectin receptor CLEC-2 signals through a pathway that is critically dependent on the tyrosine kinase Syk. We show that homozygous loss of either protein results in defects in brain vascular and lymphatic development, lung inflation, and perinatal lethality. Furthermore, we find that conditional deletion of Syk in the hematopoietic lineage, or conditional deletion of CLEC-2 or Syk in the megakaryocyte/platelet lineage, also causes defects in brain vascular and lymphatic development, although the mice are viable. In contrast, conditional deletion of Syk in other hematopoietic lineages had no effect on viability or brain vasculature and lymphatic development. We show that platelets, but not platelet releasate, modulate the migration and intercellular adhesion of lymphatic endothelial cells through a pathway that depends on CLEC-2 and Syk. These studies found that megakaryocyte/platelet expression of CLEC-2 and Syk is required for normal brain vasculature and lymphatic development and that platelet CLEC-2 and Syk directly modulate lymphatic endothelial cell behavior in vitro. (Blood. 2012;119(7):1747-1756) IntroductionRecently, several mutant mouse models have shown a defect in the separation of the lymphatic vasculature from the blood vasculature typically resulting in the appearance of blood-filled lymphatic vessels in the skin at embryonic day (E) 14.5 (review in Tammela and Alitalo 1 ). Mice deficient in the tyrosine kinase Syk show this phenotype during gestation and die around the time of birth. 2-4 A similar defect is found in mice deficient in the adapter protein SLP76 (Lcp2) 4 or in PLC␥2, 5 which play vital roles downstream of Syk in immunoreceptor tyrosine-based activation motif (ITAM) and integrin signaling cascades, providing circumstantial evidence that the Syk-SLP76-PLC␥2 pathway is required for normal lymphatic development.The C-type lectin-like protein type 2 (CLEC-2, encoded by the Clec1b gene) is highly expressed on platelets and at lower levels on other hematopoietic cells [6][7][8][9] and signals through a cytosolic YxxL sequence known as a hemITAM. 10,11 These receptors signal through a similar pathway used by ITAM receptors which have a dual YxxL/I sequence. HemITAM receptors activate Syk, initiating a signaling cascade partially dependent on SLP76 that leads to activation of PLC␥2. 6,12,13 The role of CLEC-2 in hemostasis and thrombosis is debatable because some lines of evidence suggest that it is required 14,15 and others show that it has no significant involvement in these processes. 16 CLEC-2 has been recognized as a receptor for the transmembrane protein podoplanin. 17,18 Podoplanin is expressed on lymphatic endothelial cells (LECs), lung type-1 alveolar cells, and kidney podocytes but not in blood endothelial cells (BECs). Podoplanin-deficient mice die shortly after birth because of an inability to inflate their lungs and, like Syk-deficient mice, show dilated, tortuous blood-filled lymphatics in mid-gestation. 19,20 A similar phenotype is seen in mice lacking megakaryocytes/...

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“…Pdpn associates with ezrin and regulates the activities of Rho GTPases to promote filopodia formation, cell motility, and metastasis. [152][153][154] Pdpn expression is induced by tumor promoters, including TPA, RAS, and Src, 127,155,156 and enhances transformed cell motility and invasion. 152,157 PDPN is predominantly found at the invasive front of many tumors, which is consistent with its role in promoting malignant invasion.…”

Section: Src and Downstream Chemotherapeutic Targetsmentioning

confidence: 99%

Src Points the Way to Biomarkers and Chemotherapeutic Targets

Krishnan¹,

Miller

Goldberg³

2012

Genes & Cancer

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The role of Src in tumorigenesis has been extensively studied since the work of Peyton Rous over a hundred years ago. Src is a non-receptor tyrosine kinase that plays key roles in signaling pathways controlling tumor cell growth and migration. Src regulates the activities of numerous molecules to induce cell transformation. However, transformed cells do not always migrate and realize their tumorigenic potential. They can be normalized by surrounding nontransformed cells by a process called contact normalization. Tumor cells need to override contact normalization to become malignant or metastatic. In this review, we discuss the role of Src in cell migration and contact normalization, with emphasis on Cas and Abl pathways. This paradigm illuminates several chemotherapeutic targets and may lead to the identification of new biomarkers and the development of effective anticancer treatments.

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Polarized migration of lymphatic endothelial cells is critically dependent on podoplanin regulation of Cdc42

Abstract: Navarro A, Perez RE, Rezaiekhaligh MH, Mabry SM, Ekekezie II. Polarized migration of lymphatic endothelial cells is critically dependent on podoplanin regulation of Cdc42.

Cited by 37 publications

References 39 publications

Modulation of VEGF-induced migration and network formation by lymphatic endothelial cells: Roles of platelets and podoplanin

Modulation of VEGF-induced migration and network formation by lymphatic endothelial cells: Roles of platelets and podoplanin

CLEC-2 and Syk in the megakaryocytic/platelet lineage are essential for development

Src Points the Way to Biomarkers and Chemotherapeutic Targets

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