{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,11]],"date-time":"2026-06-11T17:48:28Z","timestamp":1781200108840,"version":"3.54.1"},"reference-count":100,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2023,9,22]],"date-time":"2023-09-22T00:00:00Z","timestamp":1695340800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000274","name":"Oxford BHF Centre of Research Excellence","doi-asserted-by":"publisher","award":["RE\/08\/004"],"award-info":[{"award-number":["RE\/08\/004"]}],"id":[{"id":"10.13039\/501100000274","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000274","name":"Oxford BHF Centre of Research Excellence","doi-asserted-by":"publisher","award":["RE\/13\/1\/30181"],"award-info":[{"award-number":["RE\/13\/1\/30181"]}],"id":[{"id":"10.13039\/501100000274","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Qatar Foundation to Hamad Bin Khalifa University","award":["RE\/08\/004"],"award-info":[{"award-number":["RE\/08\/004"]}]},{"name":"Qatar Foundation to Hamad Bin Khalifa University","award":["RE\/13\/1\/30181"],"award-info":[{"award-number":["RE\/13\/1\/30181"]}]},{"name":"College of Health and Life Sciences at Hamad Bin Khalifa University","award":["RE\/08\/004"],"award-info":[{"award-number":["RE\/08\/004"]}]},{"name":"College of Health and Life Sciences at Hamad Bin Khalifa University","award":["RE\/13\/1\/30181"],"award-info":[{"award-number":["RE\/13\/1\/30181"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The impairment in microvascular network formation could delay the restoration of blood flow after acute limb ischemia. A high-content screen of a GSK-published kinase inhibitor library identified a set of ROCK inhibitor hits enhancing endothelial network formation. Subsequent kinase activity profiling against a panel of 224 protein kinases showed that two indazole-based ROCK inhibitor hits exhibited high selectivity for ROCK1 and ROCK2 isoforms compared to other ROCK inhibitors. One of the chemical entities, GSK429286, was selected for follow-up studies. We found that GSK429286 was ten times more potent in enhancing endothelial tube formation than Fasudil, a classic ROCK inhibitor. ROCK1 inhibition by RNAi phenocopied the angiogenic phenotype of the GSK429286 compound. Using an organotypic angiogenesis co-culture assay, we showed that GSK429286 formed a dense vascular network with thicker endothelial tubes. Next, mice received either vehicle or GSK429286 (10 mg\/kg i.p.) for seven days after hindlimb ischemia induction. As assessed by laser speckle contrast imaging, GSK429286 potentiated blood flow recovery after ischemia induction. At the histological level, we found that GSK429286 significantly increased the size of new microvessels in the regenerating areas of ischemic muscles compared with vehicle-treated ones. Our findings reveal that selective ROCK inhibitors have in vitro pro-angiogenic properties and therapeutic potential to restore blood flow in limb ischemia.<\/jats:p>","DOI":"10.3390\/ijms241914410","type":"journal-article","created":{"date-parts":[[2023,9,22]],"date-time":"2023-09-22T03:44:42Z","timestamp":1695354282000},"page":"14410","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Selective ROCK Inhibitor Enhances Blood Flow Recovery after Hindlimb Ischemia"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0753-9944","authenticated-orcid":false,"given":"Hend Salah","family":"Fayed","sequence":"first","affiliation":[{"name":"College of Health and Life Sciences, Hamad Bin Khalifa University, Doha P.O. Box 34110, Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mouayad Zuheir","family":"Bakleh","sequence":"additional","affiliation":[{"name":"College of Health and Life Sciences, Hamad Bin Khalifa University, Doha P.O. Box 34110, Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0889-7030","authenticated-orcid":false,"given":"Jasni Viralippurath","family":"Ashraf","sequence":"additional","affiliation":[{"name":"College of Health and Life Sciences, Hamad Bin Khalifa University, Doha P.O. Box 34110, Qatar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alison","family":"Howarth","sequence":"additional","affiliation":[{"name":"Nuffield Department of Medicine, Target Discovery Institute, University of Oxford, Oxford OX3 7FZ, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6495-7026","authenticated-orcid":false,"given":"Daniel","family":"Ebner","sequence":"additional","affiliation":[{"name":"Nuffield Department of Medicine, Target Discovery Institute, University of Oxford, Oxford OX3 7FZ, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ayman","family":"Al Haj Zen","sequence":"additional","affiliation":[{"name":"College of Health and Life Sciences, Hamad Bin Khalifa University, Doha P.O. Box 34110, Qatar"},{"name":"BHF Centre of Research Excellence, Division of Cardiovascular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford OX3 9DU, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,9,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"873","DOI":"10.1016\/j.cell.2011.08.039","article-title":"Basic and therapeutic aspects of angiogenesis","volume":"146","author":"Potente","year":"2011","journal-title":"Cell"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1038\/nm0297-158","article-title":"Angiogenesis in ischemic heart disease","volume":"3","author":"Ware","year":"1997","journal-title":"Nat. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"648","DOI":"10.1161\/ATVBAHA.108.181644","article-title":"Vascular responses to hypoxia and ischemia","volume":"30","author":"Semenza","year":"2010","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1038\/jcbfm.1990.60","article-title":"Reduction of functional capillary density in human brain after stroke","volume":"10","author":"Gjedde","year":"1990","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1186\/s12967-016-1030-6","article-title":"Walking performance is positively correlated to calf muscle fiber size in peripheral artery disease subjects, but fibers show aberrant mitophagy: An observational study","volume":"14","author":"White","year":"2016","journal-title":"J. Transl. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"444","DOI":"10.1038\/nature05602","article-title":"p53-induced inhibition of Hif-1 causes cardiac dysfunction during pressure overload","volume":"446","author":"Sano","year":"2007","journal-title":"Nature"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1177\/1358863X20945794","article-title":"Skeletal muscle capillary density is related to anaerobic threshold and claudication in peripheral artery disease","volume":"25","author":"Duscha","year":"2020","journal-title":"Vasc. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2486","DOI":"10.1096\/fj.11-197400","article-title":"Therapeutic angiogenesis due to balanced single-vector delivery of VEGF and PDGF-BB","volume":"26","author":"Banfi","year":"2012","journal-title":"FASEB J."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1944","DOI":"10.1161\/CIRCRESAHA.121.318266","article-title":"New Directions in Therapeutic Angiogenesis and Arteriogenesis in Peripheral Arterial Disease","volume":"128","author":"Annex","year":"2021","journal-title":"Circ. Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1038\/nm0603-669","article-title":"The biology of VEGF and its receptors","volume":"9","author":"Ferrara","year":"2003","journal-title":"Nat. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"464","DOI":"10.1038\/nrm2183","article-title":"Molecular regulation of angiogenesis and lymphangiogenesis","volume":"8","author":"Adams","year":"2007","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Jeltsch, M., Leppanen, V.M., Saharinen, P., and Alitalo, K. (2013). Receptor tyrosine kinase-mediated angiogenesis. Cold Spring Harb. Perspect. Biol., 5.","DOI":"10.1101\/cshperspect.a009183"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1038\/nrm2639","article-title":"Control of vascular morphogenesis and homeostasis through the angiopoietin-Tie system","volume":"10","author":"Augustin","year":"2009","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1324","DOI":"10.1158\/0008-5472.CAN-08-3030","article-title":"Host-derived angiopoietin-2 affects early stages of tumor development and vessel maturation but is dispensable for later stages of tumor growth","volume":"69","author":"Nasarre","year":"2009","journal-title":"Cancer Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1101\/gad.13.3.295","article-title":"Roles of ephrinB ligands and EphB receptors in cardiovascular development: Demarcation of arterial\/venous domains, vascular morphogenesis, and sprouting angiogenesis","volume":"13","author":"Adams","year":"1999","journal-title":"Genes Dev."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1111\/joim.12019","article-title":"VEGFA and tumour angiogenesis","volume":"273","author":"Welsh","year":"2013","journal-title":"J. Intern. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s10456-009-9160-6","article-title":"Anti-angiogenic tyrosine kinase inhibitors: What is their mechanism of action?","volume":"13","author":"Gotink","year":"2010","journal-title":"Angiogenesis"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1186\/s13045-019-0718-5","article-title":"Recent advances on anti-angiogenesis receptor tyrosine kinase inhibitors in cancer therapy","volume":"12","author":"Qin","year":"2019","journal-title":"J. Hematol. Oncol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1096\/fj.201800640RR","article-title":"Pro- and antiangiogenic therapies: Current status and clinical implications","volume":"33","author":"Rust","year":"2019","journal-title":"FASEB J."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"370","DOI":"10.1016\/S0140-6736(96)03361-2","article-title":"Clinical evidence of angiogenesis after arterial gene transfer of phVEGF165 in patient with ischaemic limb","volume":"348","author":"Isner","year":"1996","journal-title":"Lancet"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2053","DOI":"10.1016\/S0140-6736(02)08937-7","article-title":"Therapeutic angiogenesis with recombinant fibroblast growth factor-2 for intermittent claudication (the TRAFFIC study): A randomised trial","volume":"359","author":"Lederman","year":"2002","journal-title":"Lancet"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"713","DOI":"10.1161\/ATVBAHA.110.219550","article-title":"Phase I\/IIa clinical trial of therapeutic angiogenesis using hepatocyte growth factor gene transfer to treat critical limb ischemia","volume":"31","author":"Morishita","year":"2011","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1177\/1753944718786926","article-title":"Results of 5-year follow-up study in patients with peripheral artery disease treated with PL-VEGF165 for intermittent claudication","volume":"12","author":"Deev","year":"2018","journal-title":"Ther. Adv. Cardiovasc. Dis."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1177\/1074248415574336","article-title":"pCMV-vegf165 Intramuscular Gene Transfer is an Effective Method of Treatment for Patients with Chronic Lower Limb Ischemia","volume":"20","author":"Deev","year":"2015","journal-title":"J. Cardiovasc. Pharmacol. Ther."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Beltran-Camacho, L., Rojas-Torres, M., and Duran-Ruiz, M.C. (2021). Current Status of Angiogenic Cell Therapy and Related Strategies Applied in Critical Limb Ischemia. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22052335"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1016\/j.tcm.2004.09.001","article-title":"Gene therapy for ischemic cardiovascular diseases: Some lessons learned from the first clinical trials","volume":"14","author":"Markkanen","year":"2004","journal-title":"Trends Cardiovasc. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1161\/CIRCULATIONAHA.108.784371","article-title":"Therapeutic angiogenesis for critical limb ischemia: Microvascular therapies coming of age","volume":"118","author":"Tongers","year":"2008","journal-title":"Circulation"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1006\/mthe.2002.0638","article-title":"Increased vascularity detected by digital subtraction angiography after VEGF gene transfer to human lower limb artery: A randomized, placebo-controlled, double-blinded phase II study","volume":"6","author":"Makinen","year":"2002","journal-title":"Mol. Ther."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1933","DOI":"10.1161\/01.CIR.0000093398.16124.29","article-title":"Regional angiogenesis with vascular endothelial growth factor in peripheral arterial disease: A phase II randomized, double-blind, controlled study of adenoviral delivery of vascular endothelial growth factor 121 in patients with disabling intermittent claudication","volume":"108","author":"Rajagopalan","year":"2003","journal-title":"Circulation"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"863","DOI":"10.1038\/nrd1226","article-title":"Therapeutic angiogenesis in cardiovascular disease","volume":"2","author":"Simons","year":"2003","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1038\/nbt.3374","article-title":"Comprehensive characterization of the Published Kinase Inhibitor Set","volume":"34","author":"Elkins","year":"2016","journal-title":"Nat. Biotechnol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1517\/17460440903493431","article-title":"Capillary microfluidic electrophoretic mobility shift assays: Application to enzymatic assays in drug discovery","volume":"5","author":"Perrin","year":"2010","journal-title":"Expert. Opin. Drug Discov."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Rudolf, A.F., Skovgaard, T., Knapp, S., Jensen, L.J., and Berthelsen, J. (2014). A comparison of protein kinases inhibitor screening methods using both enzymatic activity and binding affinity determination. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0098800"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1021\/jm0609014","article-title":"Development of dihydropyridone indazole amides as selective Rho-kinase inhibitors","volume":"50","author":"Goodman","year":"2007","journal-title":"J. Med. Chem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"782","DOI":"10.1161\/01.RES.0000259593.07661.1e","article-title":"Endothelial cell migration during angiogenesis","volume":"100","author":"Lamalice","year":"2007","journal-title":"Circ. Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3599","DOI":"10.1074\/mcp.M113.031344","article-title":"SILAC-based proteomics of human primary endothelial cell morphogenesis unveils tumor angiogenic markers","volume":"12","author":"Zanivan","year":"2013","journal-title":"Mol. Cell. Proteom."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Mongiat, M., Andreuzzi, E., Tarticchio, G., and Paulitti, A. (2016). Extracellular Matrix, a Hard Player in Angiogenesis. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17111822"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"23766","DOI":"10.1074\/jbc.M311675200","article-title":"Laminin-1 promotes angiogenesis in synergy with fibroblast growth factor by distinct regulation of the gene and protein expression profile in endothelial cells","volume":"279","author":"Dixelius","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"747","DOI":"10.3389\/fimmu.2019.00747","article-title":"Myosin II Activity Is Selectively Needed for Migration in Highly Confined Microenvironments in Mature Dendritic Cells","volume":"10","author":"Barbier","year":"2019","journal-title":"Front. Immunol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"e483","DOI":"10.1038\/cddis.2013.10","article-title":"Distinct roles for ROCK1 and ROCK2 in the regulation of cell detachment","volume":"4","author":"Shi","year":"2013","journal-title":"Cell Death Dis."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"276","DOI":"10.3389\/fphar.2015.00276","article-title":"The Function of Rho-Associated Kinases ROCK1 and ROCK2 in the Pathogenesis of Cardiovascular Disease","volume":"6","author":"Hartmann","year":"2015","journal-title":"Front. Pharmacol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1007\/978-1-4939-9585-1_5","article-title":"In Vitro Models to Study the Regulatory Roles of Retinoids in Angiogenesis","volume":"2019","year":"2019","journal-title":"Methods Mol. Biol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1002\/cyto.a.20808","article-title":"A novel imaging-based high-throughput screening approach to anti-angiogenic drug discovery","volume":"77","author":"Evensen","year":"2010","journal-title":"Cytometry A"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"24266","DOI":"10.1074\/jbc.273.37.24266","article-title":"Post-transcriptional regulation of endothelial nitric oxide synthase mRNA stability by Rho GTPase","volume":"273","author":"Laufs","year":"1998","journal-title":"J. Biol. Chem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"8467","DOI":"10.1128\/MCB.22.24.8467-8477.2002","article-title":"Rho GTPase\/Rho kinase negatively regulates endothelial nitric oxide synthase phosphorylation through the inhibition of protein kinase B\/Akt in human endothelial cells","volume":"22","author":"Ming","year":"2002","journal-title":"Mol. Cell Biol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1161\/01.CIR.93.1.85","article-title":"L-arginine induces nitric oxide-dependent vasodilation in patients with critical limb ischemia. A randomized, controlled study","volume":"93","author":"Boger","year":"1996","journal-title":"Circulation"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"10999","DOI":"10.1073\/pnas.0501444102","article-title":"Endothelial nitric oxide synthase is critical for ischemic remodeling, mural cell recruitment, and blood flow reserve","volume":"102","author":"Yu","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1161\/CIRCRESAHA.110.221663","article-title":"Inhibition of delta-like-4-mediated signaling impairs reparative angiogenesis after ischemia","volume":"107","author":"Oikawa","year":"2010","journal-title":"Circ. Res."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1007\/s10456-009-9146-4","article-title":"The endothelial cell tube formation assay on basement membrane turns 20: State of the science and the art","volume":"12","author":"Arnaoutova","year":"2009","journal-title":"Angiogenesis"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"628","DOI":"10.1038\/nprot.2010.6","article-title":"In vitro angiogenesis: Endothelial cell tube formation on gelled basement membrane extract","volume":"5","author":"Arnaoutova","year":"2010","journal-title":"Nat. Protoc."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1745","DOI":"10.1038\/mt.2016.153","article-title":"The Retinoid Agonist Tazarotene Promotes Angiogenesis and Wound Healing","volume":"24","author":"Nawrot","year":"2016","journal-title":"Mol. Ther."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1021\/jm060873p","article-title":"Discovery of aminofurazan-azabenzimidazoles as inhibitors of Rho-kinase with high kinase selectivity and antihypertensive activity","volume":"50","author":"Stavenger","year":"2007","journal-title":"J. Med. Chem."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"25228","DOI":"10.1039\/D1RA03979B","article-title":"Current progress, challenges and future prospects of indazoles as protein kinase inhibitors for the treatment of cancer","volume":"11","author":"Tandon","year":"2021","journal-title":"RSC Adv."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"H893","DOI":"10.1152\/ajpheart.01038.2008","article-title":"Inhibition of Rho-dependent kinases ROCK I\/II activates VEGF-driven retinal neovascularization and sprouting angiogenesis","volume":"296","author":"Kroll","year":"2009","journal-title":"Am. J. Physiol. Heart Circ. Physiol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1002\/cm.20472","article-title":"Rho-kinase\/ROCK: A key regulator of the cytoskeleton and cell polarity","volume":"67","author":"Amano","year":"2010","journal-title":"Cytoskeleton"},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Pipparelli, A., Arsenijevic, Y., Thuret, G., Gain, P., Nicolas, M., and Majo, F. (2013). ROCK inhibitor enhances adhesion and wound healing of human corneal endothelial cells. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0062095"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1016\/j.cub.2008.12.045","article-title":"Local cortical tension by myosin II guides 3D endothelial cell branching","volume":"19","author":"Fischer","year":"2009","journal-title":"Curr. Biol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1038\/ncb3092","article-title":"Myosin II controls cellular branching morphogenesis and migration in three dimensions by minimizing cell-surface curvature","volume":"17","author":"Elliott","year":"2015","journal-title":"Nat. Cell Biol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1038\/nrm2957","article-title":"Cell adhesion: Integrating cytoskeletal dynamics and cellular tension","volume":"11","author":"Parsons","year":"2010","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"3186","DOI":"10.1096\/fj.09-145102","article-title":"RhoA\/ROCK signaling is essential for multiple aspects of VEGF-mediated angiogenesis","volume":"24","author":"Bryan","year":"2010","journal-title":"FASEB J."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1159\/000492994","article-title":"Distinct Roles For ROCK1 and ROCK2 in the Regulation of Oxldl-Mediated Endothelial Dysfunction","volume":"49","author":"Huang","year":"2018","journal-title":"Cell Physiol. Biochem."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"446","DOI":"10.1038\/nrm1128","article-title":"Rocks: Multifunctional kinases in cell behaviour","volume":"4","author":"Riento","year":"2003","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1586\/14779072.3.3.441","article-title":"ROCKs as therapeutic targets in cardiovascular diseases","volume":"3","author":"Rikitake","year":"2005","journal-title":"Expert Rev. Cardiovasc. Ther."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"5043","DOI":"10.1128\/MCB.23.14.5043-5055.2003","article-title":"Targeted disruption of the mouse rho-associated kinase 2 gene results in intrauterine growth retardation and fetal death","volume":"23","author":"Thumkeo","year":"2003","journal-title":"Mol. Cell Biol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"941","DOI":"10.1083\/jcb.200411179","article-title":"ROCK-I regulates closure of the eyelids and ventral body wall by inducing assembly of actomyosin bundles","volume":"168","author":"Shimizu","year":"2005","journal-title":"J. Cell Biol."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"1029","DOI":"10.1016\/S0008-6363(99)00144-3","article-title":"Rho-kinase-mediated pathway induces enhanced myosin light chain phosphorylations in a swine model of coronary artery spasm","volume":"43","author":"Shimokawa","year":"1999","journal-title":"Cardiovasc. Res."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1161\/01.RES.87.3.195","article-title":"Involvement of Rho-kinase-mediated phosphorylation of myosin light chain in enhancement of cerebral vasospasm","volume":"87","author":"Sato","year":"2000","journal-title":"Circ. Res."},{"key":"ref_68","first-page":"1062","article-title":"Involvement of Rho-kinase in hypertensive vascular disease: A novel therapeutic target in hypertension","volume":"15","author":"Mukai","year":"2001","journal-title":"FASEB J."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1161\/01.RES.0000111804.34509.94","article-title":"Long-term treatment with a Rho-kinase inhibitor improves monocrotaline-induced fatal pulmonary hypertension in rats","volume":"94","author":"Abe","year":"2004","journal-title":"Circ. Res."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"1441","DOI":"10.1016\/S0024-3205(01)01229-2","article-title":"Pharmacological profile of hydroxy fasudil as a selective rho kinase inhibitor on ischemic brain damage","volume":"69","author":"Satoh","year":"2001","journal-title":"Life Sci."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"421","DOI":"10.2176\/nmc.46.421","article-title":"Effect of fasudil hydrochloride, a protein kinase inhibitor, on cerebral vasospasm and delayed cerebral ischemic symptoms after aneurysmal subarachnoid hemorrhage","volume":"46","author":"Zhao","year":"2006","journal-title":"Neurol. Med. Chir."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1097\/FJC.0b013e318070d1bd","article-title":"Rho kinase (ROCK) inhibitors","volume":"50","author":"Liao","year":"2007","journal-title":"J. Cardiovasc. Pharmacol."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"810","DOI":"10.1161\/CIRCRESAHA.116.309094","article-title":"Rasip1-Mediated Rho GTPase Signaling Regulates Blood Vessel Tubulogenesis via Nonmuscle Myosin II","volume":"119","author":"Barry","year":"2016","journal-title":"Circ. Res."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1016\/0040-8166(86)90026-1","article-title":"The pericyte\u2014A review","volume":"18","author":"Sims","year":"1986","journal-title":"Tissue Cell"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"700","DOI":"10.1038\/nature05193","article-title":"Bidirectional control of CNS capillary diameter by pericytes","volume":"443","author":"Peppiatt","year":"2006","journal-title":"Nature"},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1146\/annurev-physiol-061121-040127","article-title":"Pericyte Control of Blood Flow Across Microvascular Zones in the Central Nervous System","volume":"84","author":"Hartmann","year":"2022","journal-title":"Annu. Rev. Physiol."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1161\/CIRCRESAHA.108.188524","article-title":"ROCK isoform regulation of myosin phosphatase and contractility in vascular smooth muscle cells","volume":"104","author":"Wang","year":"2009","journal-title":"Circ. Res."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"5341","DOI":"10.7150\/thno.42077","article-title":"Direct myosin-2 inhibition enhances cerebral perfusion resulting in functional improvement after ischemic stroke","volume":"10","author":"Penzes","year":"2020","journal-title":"Theranostics"},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1016\/j.phrs.2018.05.011","article-title":"RhoA\/ROCK signaling regulates smooth muscle phenotypic modulation and vascular remodeling via the JNK pathway and vimentin cytoskeleton","volume":"133","author":"Tang","year":"2018","journal-title":"Pharmacol. Res."},{"key":"ref_80","doi-asserted-by":"crossref","unstructured":"Ashraf, J.V., and Al Haj Zen, A. (2021). Role of Vascular Smooth Muscle Cell Phenotype Switching in Arteriogenesis. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms221910585"},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1124\/jpet.106.110635","article-title":"Novel Rho kinase inhibitors with anti-inflammatory and vasodilatory activities","volume":"320","author":"Doe","year":"2007","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1093\/cvr\/cvs371","article-title":"Prevention of diabetes-induced arginase activation and vascular dysfunction by Rho kinase (ROCK) knockout","volume":"97","author":"Yao","year":"2013","journal-title":"Cardiovasc. Res."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/S1072-7515(98)00259-2","article-title":"Protective role of nitric oxide in ischemia and reperfusion injury of the liver","volume":"188","author":"Shimamura","year":"1999","journal-title":"J. Am. Coll. Surg."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1021\/cb300610s","article-title":"Kinase drug discovery\u2014What\u2019s next in the field?","volume":"8","author":"Cohen","year":"2013","journal-title":"ACS Chem. Biol."},{"key":"ref_85","doi-asserted-by":"crossref","unstructured":"Aref, Z., de Vries, M.R., and Quax, P.H.A. (2019). Variations in Surgical Procedures for Inducing Hind Limb Ischemia in Mice and the Impact of These Variations on Neovascularization Assessment. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20153704"},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"1546","DOI":"10.1016\/j.jvs.2008.07.063","article-title":"Cellular and molecular mechanism regulating blood flow recovery in acute versus gradual femoral artery occlusion are distinct in the mouse","volume":"48","author":"Yang","year":"2008","journal-title":"J. Vasc. Surg."},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"340","DOI":"10.2174\/1568026613666131127160819","article-title":"Seeding collaborations to advance kinase science with the GSK Published Kinase Inhibitor Set (PKIS)","volume":"14","author":"Drewry","year":"2014","journal-title":"Curr. Top Med. Chem."},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1177\/108705719900400206","article-title":"A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays","volume":"4","author":"Zhang","year":"1999","journal-title":"J. Biomol. Screen."},{"key":"ref_89","doi-asserted-by":"crossref","unstructured":"Pelz, O., Gilsdorf, M., and Boutros, M. (2010). web cellHTS2: A web-application for the analysis of high-throughput screening data. BMC Bioinform., 11.","DOI":"10.1186\/1471-2105-11-185"},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"3713","DOI":"10.1158\/0008-5472.CAN-08-2515","article-title":"Combined integrin phosphoproteomic analyses and small interfering RNA-based functional screening identify key regulators for cancer cell adhesion and migration","volume":"69","author":"Chen","year":"2009","journal-title":"Cancer Res."},{"key":"ref_91","doi-asserted-by":"crossref","first-page":"3090","DOI":"10.1074\/jbc.273.5.3090","article-title":"Adhesion of human umbilical vein endothelial cells to the immediate-early gene product Cyr61 is mediated through integrin alphavbeta3","volume":"273","author":"Kireeva","year":"1998","journal-title":"J. Biol. Chem."},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"4565","DOI":"10.1182\/blood-2011-09-380006","article-title":"Blocking ephrinB2 with highly specific antibodies inhibits angiogenesis, lymphangiogenesis, and tumor growth","volume":"119","author":"Abengozar","year":"2012","journal-title":"Blood"},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"eadd2488","DOI":"10.1126\/sciadv.add2488","article-title":"A self-generated Toddler gradient guides mesodermal cell migration","volume":"8","author":"Stock","year":"2022","journal-title":"Sci. Adv."},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"11568","DOI":"10.1038\/s41598-020-67289-8","article-title":"Angiogenesis Analyzer for ImageJ\u2014A comparative morphometric analysis of \u201cEndothelial Tube Formation Assay\u201d and \u201cFibrin Bead Assay\u201d","volume":"10","author":"Carpentier","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_95","first-page":"1667","article-title":"Mouse model of angiogenesis","volume":"152","author":"Couffinhal","year":"1998","journal-title":"Am. J. Pathol."},{"key":"ref_96","doi-asserted-by":"crossref","first-page":"2251","DOI":"10.1161\/01.STR.0000181077.84981.11","article-title":"Inhibition of Rho kinase (ROCK) leads to increased cerebral blood flow and stroke protection","volume":"36","author":"Rikitake","year":"2005","journal-title":"Stroke"},{"key":"ref_97","doi-asserted-by":"crossref","first-page":"13651","DOI":"10.1038\/s41598-020-70391-6","article-title":"Dysregulation of ghrelin in diabetes impairs the vascular reparative response to hindlimb ischemia in a mouse model; clinical relevance to peripheral artery disease","volume":"10","author":"Neale","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_98","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.omtn.2018.08.006","article-title":"Phenotypic miRNA Screen Identifies miR-26b to Promote the Growth and Survival of Endothelial Cells","volume":"13","author":"Martello","year":"2018","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_99","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/S0014-5793(01)02901-5","article-title":"Reliable in vitro measurement of nitric oxide released from endothelial cells using low concentrations of the fluorescent probe 4,5-diaminofluorescein","volume":"506","author":"Leikert","year":"2001","journal-title":"FEBS Lett."},{"key":"ref_100","first-page":"1195","article-title":"Echinacoside-induced nitric oxide production in endothelial cells: Roles of androgen receptor and the PI3K-Akt pathway","volume":"45","author":"Gu","year":"2020","journal-title":"Int. J. Mol. Med."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/19\/14410\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T20:55:26Z","timestamp":1760129726000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/19\/14410"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,9,22]]},"references-count":100,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2023,10]]}},"alternative-id":["ijms241914410"],"URL":"https:\/\/doi.org\/10.3390\/ijms241914410","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,9,22]]}}}