Role of ROCK I/II in vascular branching
2009; American Physical Society; Volume: 296; Issue: 4 Linguagem: Inglês
10.1152/ajpheart.00125.2009
ISSN1522-1539
AutoresGeerten P. van Nieuw Amerongen, Victor W.M. van Hinsbergh,
Tópico(s)Intracerebral and Subarachnoid Hemorrhage Research
ResumoEDITORIAL FOCUSRole of ROCK I/II in vascular branchingGeerten P. van Nieuw Amerongen and Victor W. M. van HinsberghGeerten P. van Nieuw Amerongen and Victor W. M. van HinsberghPublished Online:01 Apr 2009https://doi.org/10.1152/ajpheart.00125.2009This is the final version - click for previous versionMoreSectionsPDF (100 KB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInWeChat the highly similar rho kinases ROCK I and II have drawn a lot of attention over the past decade as potential targets for novel treatment options for a variety of cardiovascular disorders such as (pulmonary) hypertension, ischemic stroke, and vascular leakage (27). Several previous in vivo studies suggested that inhibition of Rho kinase also could provide a potent strategy for inhibition of angiogenesis (9, 21, 22, 26, 28, 31, 37). Data obtained from in vitro and explant models, however, indicated that Rho kinase inhibitors do not only impair angiogenesis but also may enhance in vitro tube formation (19, 29). In their article, Kroll et al. (15) provide important in vivo evidence for a potentiating effect of Rho kinase inhibition on angiogenesis, supplying further fuel to this controversial issue (Fig. 1). Together with a recent report by Fischer et al. (5), these data shed new light on the role of Rho kinase in angiogenesis. Novel information on the contribution of disordered RhoA activity to excessive, but dysfunctional, angiogenesis in a common vascular dysplasia called cerebral cavernous malformation (CCM) underscores the importance of these findings (36).Fig. 1.Proposed dual role for Rho-dependent kinase (ROCK) I/II in regulation of angiogenesis. See text for details.Download figureDownload PowerPointThe role of members of the Rho family of small GTPases, with its main members RhoA, Rac1, and Cdc42, as key regulators of angiogenesis has been well established [see Bryan and d'Amore (3) for review]. Rho GTPases modulate a diversity of cellular processes, including extracellular matrix (ECM) remodeling, migration, proliferation, morphogenesis, and survival. RhoA signaling plays an essential role in vascular endothelial growth factor (VEGF)-dependent in vivo angiogenesis and in initial steps of in vitro endothelial cord assembly (3, 11, 23, 34). RhoC has also been implicated (35). Other studies have suggested that RhoA plays a role in endothelial tube collapse and regression (4). Using a RNAi-mediated suppression approach, Cdc42 and Rac1 have been implicated in the process of endothelial lumen formation, whereas RhoA appears to play a minimal role here (14).The effects of RhoA are mediated at least in part by its downstream target Rho kinase. Where previous studies with in vitro or explant and in vivo models indicated an essential role for Rho kinase in angiogenesis, we provided in 2003 the first evidence for a dual role of Rho kinase in angiogenesis (32); the Rho kinase inhibitor Y-27632 reduced the length of VEGF-induced tube-like structures in an in vitro model for angiogenesis, but Y-27632 also enhanced initial sprouting. These data suggested a role for Rho kinase in three-dimensional migration, which was confirmed in a two-dimensional wound healing model. However, the increased sprouting upon treatment with the Rho kinase inhibitor remained unexplained at that time.The merit of the work of Kroll et al. is that they elucidate a putative inhibitory role of Rho kinase in VEGF-driven angiogenesis, providing in vivo evidence for an inhibitory role for Rho kinase in neovascularization. In a mouse retinal vascularization model, they show that a highly selective Rho kinase inhibitor enhanced angiogenesis by ∼50%. Similarly, downregulation of ROCK I/II expression in cultured endothelial cells (ECs) enhanced tube formation in a human umbilical vein endothelial cell spheroid model. Furthermore, they provide a novel molecular explanation for the counteracting role of Rho kinase by demonstrating that Rho kinase inhibitors enhance activation of the VEGF receptor VEGFR2/KDR. It remains to be investigated whether this VEGFR2 hyperactivation acts synergistically with enhanced nitric oxide (NO) bioavailability, since inhibition of Rho kinase is known to induce expression of endothelial nitric oxide synthase and NO plays an integral role in development and maintenance of the microvascular network (25).An important feature in determining the morphology of tubular systems such as the vascular bed is the frequency and geometry of branching. Hence, deciphering the molecular mechanisms underlying the sprouting of new branches is the key to understanding the formation of tubular systems (12). In recent years, much attention has been directed toward the role played by tip cells in the growth of blood vessels. Much of the knowledge of endothelial tip vs. stalk cell specification in the vascular system was derived from observations of angiogenesis in the postnatal mouse retina (12). An important molecular mechanism is induction and expression of delta-like ligand 4 (dll4) by VEGFR activation in individual cells conferring a cell-tip phenotype and activating Notch in adjacent ECs. Notch activation suppresses VEGFR expression and prevents these cells from conversion into tip cells (10, 17). Interestingly, dll4+/− mice have a phenotype highly reminiscent to that induced by inhibition of Rho kinase as described by Fischer et al. (5), showing an increased number of sprouting vessels. Data from keratinocytes and vascular smooth muscle cells indicate that Notch signaling interferes both with ROCK I/II expression and activity (2, 16), indeed suggesting a link between Notch signaling and Rho kinase activity.Fischer et al. (5) now identify ROCK-mediated myosin II activity and stiffness of the ECM as two important cues inhibiting endothelial pseudopodial branch initiation, i.e., inhibition of ROCK I/II or reduced ECM stiffness promote branching. Myosin II is dynamically localized to the endothelial cortex and is partially released under conditions that promote branching. Local downregulation of myosin II-mediated cortical contraction allows pseudopodium initiation to mediate endothelial branching and hence guide directional migration and angiogenesis. Two recent studies indicate that aberrant RhoA/Rho kinase signaling contributes to pathological forms of angiogenesis (7, 36). 1) Targeted disruption of the gene that causes cerebral cavernous malformation, Ccm2, hyperactivates RhoA, resulting in excessive but dysfunctional angiogenic sprouts form, which are unable to develop stable lumens to allow functional circulation. Normalization of RhoA activity with statins rescues the vasculature. 2) Tumor capillary ECs more readily form capillary networks in vitro because of enhanced Rho kinase activity (7). These cells exert greater traction force and display an enhanced ability to retract flexible ECM substrates. Moreover, decreasing Rho-mediated tension by the ROCK inhibitor Y-27632 can reprogram the tumor capillary ECs so that they normalize their ability to form tubular networks on ECM gels.It is increasingly appreciated that the GTP-binding-GTP-hydrolysis cycle of Rho GTPases is highly coordinated in a spatiotemporally controlled manner for effective signaling output (30). During angiogenesis, specific guanine exchange factors (RhoGEFs) and GTPase-activating proteins (RhoGAPs) are dedicated to regulate RhoA activity. Loss-of-function studies in the zebrafish and mouse point to a specific role for the RhoGEF Syx in angiogenic sprouting in the developing vascular bed (6). Importantly, vasculogenesis and angioblast differentiation steps were unaffected. A vascular cell-restricted RhoGAP, p73-RhoGAP, plays a key role in angiogenesis (29), and, recently, p190B-RhoGAP also has been implicated in regulation of angiogenesis (8). Taken together, these data suggest that localized RhoA/Rho kinase activities determine branching sites of tip cells by regulating cortical contraction, whereas RhoA activity localized to the trailing edge facilitates migration of stalk cells by promoting tail retraction.In one of their previous studies using the same spheroid model, Nacak et al. (20) implicated the RhoGEF ECT-2 in regulation of angiogenesis, demonstrating that VEGF-induced activation of RhoA is indispensable for angiogenesis. It remains hard to reconcile these data with the current study, where they find enhanced angiogenesis when Rho kinase activity is abolished. It suggests that at least part of the effect of RhoA is mediated by downstream targets of RhoA other than ROCK I/II. RhoA-independent activation of ROCK, however, cannot be excluded, since Rho kinase also can be activated by other Rho proteins such as RhoB and RhoC, by arachidonic acid, by proteolytic cleavage by caspases, by phosphorylation by polo-like kinase-1, or even by methylation or altered expression such as induced by Notch (18, 24). An attractive alternative explanation might be that, in the models used by Kroll et al., the angiogenesis-inhibitory ROCK activities outweigh the angiogenesis-promoting ROCK activities. It would be interesting to investigate whether these angiogenesis-promoting and -inhibiting activities of Rho kinase could be attributed to specific ROCK isoforms, since studies are accumulating that show that ROCK I and ROCK II regulate different aspects of myosin II activity, resulting in distinct biological functions of ROCK I and II (33, 38).Although the finding that ROCK I/II inhibitors promote angiogenesis fits with the general idea that inhibition of Rho kinase has positive effects on the vasculature, from a therapeutic point of view, this raises concern to an unlimited use of ROCK I/II inhibitors, especially in the eye. Rho kinase has been proposed as a unique therapeutic target in the treatment of proliferative vitreoretinal diseases (13) and diabetic retinopathy (1). Because disordered angiogenesis causes blindness, careful clinical investigation is warranted. Perhaps isoform-selective inhibitors may offer a solution here.GRANTSG. 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Medical Center, Institute for Cardiovascular Research, Dept. for Physiology, van der Boechorststraat 7, 1081BT Amsterdam, The Netherlands (e-mail: [email protected]) Download PDF Back to Top Next FiguresReferencesRelatedInformationCited ByPossible cooption of a VEGF-driven tubulogenesis program for biomineralization in echinoderms31 May 2019 | Proceedings of the National Academy of Sciences, Vol. 116, No. 25Effects of ripasudil, a ROCK inhibitor, on retinal edema and nonperfusion area in a retinal vein occlusion murine modelJournal of Pharmacological Sciences, Vol. 137, No. 2AMA0428, A Potent Rock Inhibitor, Attenuates Early and Late Experimental Diabetic Retinopathy11 July 2016 | Current Eye Research, Vol. 42, No. 2VASCULAR ENDOTHELIAL GROWTH FACTOR IN HEALTH AND DISEASE: A REVIEW6 October 2016 | Journal of Evidence Based Medicine and Healthcare, Vol. 3, No. 80Morphogenesis of 3D vascular networks is regulated by tensile forces7 March 2016 | Proceedings of the National Academy of Sciences, Vol. 113, No. 12Genome-wide Approaches Reveal Functional Vascular Endothelial Growth Factor (VEGF)-inducible Nuclear Factor of Activated T Cells (NFAT) c1 Binding to Angiogenesis-related Genes in the EndotheliumJournal of Biological Chemistry, Vol. 289, No. 42Other Major Types of Signaling Mediators4 July 2012Signaling Pathways4 July 2012Conclusion4 July 2012Signaling Lipids4 July 2012Preamble to Cytoplasmic Protein Kinases4 July 2012Cytoplasmic Protein Tyrosine Kinases4 July 2012Cytoplasmic Protein Serine/Threonine Kinases4 July 2012Mitogen-Activated Protein Kinase Module4 July 2012Dual-Specificity Protein Kinases4 July 2012Cytosolic Protein Phosphatases4 July 2012Guanosine Triphosphatases and Their Regulators4 July 2012Shroom2 regulates contractility to control endothelial morphogenesisMolecular Biology of the Cell, Vol. 22, No. 6PI3K Signaling Through the Dual GTPase–Activating Protein ARAP3 Is Essential for Developmental AngiogenesisScience Signaling, Vol. 3, No. 145 More from this issue > Volume 296Issue 4April 2009Pages H903-H905 Copyright & PermissionsCopyright © 2009 the American Physiological Societyhttps://doi.org/10.1152/ajpheart.00125.2009PubMed19218507History Published online 1 April 2009 Published in print 1 April 2009 Metrics
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