Integrin α2β1 mediates outside-in regulation of platelet spreading on collagen through activation of Src kinases and PLCγ2

Collagen plays a critical role in hemostasis by promoting adhesion and activation of platelets at sites of vessel injury. In the present model of platelet–collagen interaction, adhesion is mediated via the inside-out regulation of integrin α2β1 and activation through the glycoprotein VI (GPVI)–Fc receptor (FcR) γ-chain complex. The present study extends this model by demonstrating that engagement of α2β1 by an integrin-specific sequence from within collagen or by collagen itself generates tyrosine kinase–based intracellular signals that lead to formation of filopodia and lamellipodia in the absence of the GPVI–FcR γ-chain complex. The same events do not occur in platelet suspensions. α2β1 activation of adherent platelets stimulates tyrosine phosphorylation of many of the proteins in the GPVI–FcR γ-chain cascade, including Src, Syk, SLP-76, and PLCγ2 as well as plasma membrane calcium ATPase and focal adhesion kinase. α2β1-mediated spreading is dramatically inhibited in the presence of the Src kinase inhibitor PP2 and in PLCγ2-deficient platelets. Spreading is abolished by chelation of intracellular Ca2+. Demonstration that adhesion of platelets to collagen via α2β1 generates intracellular signals provides a new insight into the mechanisms that control thrombus formation and may explain the unstable nature of β1-deficient thrombi and why loss of the GPVI–FcR γ-chain complex has a relatively minor effect on bleeding

Roles of the CLEC-2–podoplanin interaction in tumor progression

Podoplanin is a type-I transmembrane sialomucin-like glycoprotein expressed on the surface of several kinds of tumor cells. The podoplanin receptor is a platelet activation receptor known as C-type lectin-like receptor 2 (CLEC-2), which has been identified as a receptor for the platelet-activating snake venom protein rhodocytin. CLEC-2 is highly expressed in platelets and megakaryocytes and expressed at lower levels in liver Kupffer cells. Podoplanin is expressed in certain types of tumor cells, including squamous cell carcinomas, seminomas, and brain tumors. Podoplanin is also expressed in a wide range of normal cells, including fibroblastic reticular cells in lymph nodes, kidney podocytes, and lymphatic endothelial cells, but not vascular endothelial cells. Metastasis of podoplanin-positive lung tumors injected from the tail vein is greatly inhibited in CLEC-2-depleted mice or in anti-podoplanin antibody-treated mice. These findings suggest that the CLEC-2–podoplanin interaction facilitates hematogenous tumor metastasis. Platelets may increase the survival of tumor cells by covering tumor cells and physically protecting them from shear stress or immune cells in the bloodstream. Alternatively, platelets may stimulate the epithelial–mesenchymal transition of tumor cells to facilitate their extravasation from blood vessels. Cell proliferation is stimulated in podoplanin-expressing tumor cells by the coculture with platelets, but the effects of the CLEC-2–podoplanin interaction on tumor growth in vivo are not yet resolved. It is possible that the CLEC-2–podoplanin interaction facilitates tumor-related thrombosis, subsequent inflammation, inflammation-induced cachexia, and reduced survival. Considering these findings, anti-podoplanin and anti-CLEC-2 drugs are promising therapies for the prevention of tumor metastasis, progression, and tumor-related symptoms, which may result in longer survival in cancer patients. There are advantages and disadvantages of anti-podoplanin vs. anti-CLEC-2 therapy. Side effects in podoplanin-expressing normal tissues due to treatment with anti-podoplanin and temporal thrombocytopenia due to treatment with anti-CLEC2 are potential problems, although solutions to these problems have been reported