{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T12:07:36Z","timestamp":1788782856296,"version":"build-2803163510"},"reference-count":43,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T00:00:00Z","timestamp":1788739200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the HIGO Program, Kumamoto University Advanced Research Project Stem Cell-Based Tissue Regeneration Research and Education Unit"},{"id":[{"id":"https:\/\/ror.org\/02cgss904","id-type":"ROR","asserted-by":"publisher"}]},{"name":"the Joint Usage\/Research Center for Developmental Medicine, IMEG, Kumamoto University"},{"id":[{"id":"https:\/\/ror.org\/02cgss904","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001695","name":"JST","doi-asserted-by":"publisher","award":["JPMJSP2136"],"award-info":[{"award-number":["JPMJSP2136"]}],"id":[{"id":"10.13039\/501100001695","id-type":"DOI","asserted-by":"publisher"}]},{"name":"the Sasakawa Scientific Research Grant from the Japan Science Society"},{"id":[{"id":"https:\/\/ror.org\/03aeyg892","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J. Dev. Biol.","JDB"],"abstract":"<jats:p>Hemostasis and wound healing are highly coordinated processes that involve rapid clot formation followed by controlled remodeling of the injured tissue microenvironment. Prior work on Akhirin (AKH), a secreted extracellular matrix protein containing two von Willebrand factor A domains and an LCCL domain, has established it as a regulator of the neural stem niche in the developing brain and spinal cord injury microenvironment. However, its role as a non-hematopoietic molecule in the vascular injury response remains unknown. Here, we present evidence that AKH contributes to hemostasis and wound repair outside the neural niche. Our immunohistochemical and biochemical analyses demonstrated AKH around arterial tissues, suggesting a potential role at the blood-vessel interface. AKH-deficient mice exhibited a striking phenotype characterized by prolonged tail bleeding and delayed wound closure, indicating impaired vascular injury repair in vivo. Furthermore, bone marrow transplantation failed to rescue the prolonged bleeding phenotype, supporting a predominant non-hematopoietic contribution. Intriguingly, analysis of classical coagulation revealed an apparent paradox: activated partial thromboplastin time was shortened, whereas prothrombin time was not significantly altered. In contrast, increased expression of tissue plasminogen activator, urokinase-type plasminogen activator, and urokinase-type plasminogen activator receptor in AKH-deficient samples suggested dysregulated local fibrinolytic remodeling. Together, these findings identify AKH as a previously unrecognized extracellular regulator of hemostatic wound repair. Rather than indicating a defect in classical coagulation cascade activation, the findings associate AKH deficiency with impaired hemostatic control and altered expression of plasminogen activator system components, suggesting a role for AKH in the local vascular injury response.<\/jats:p>","DOI":"10.3390\/jdb14030041","type":"journal-article","created":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T10:30:46Z","timestamp":1788777046000},"page":"41","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Akhirin Preserves Hemostatic Wound Repair Through Non-Hematopoietic Regulation of the Vascular Injury Microenvironment"],"prefix":"10.3390","volume":"14","author":[{"given":"Mohammad Badrul","family":"Anam","sequence":"first","affiliation":[{"name":"Department of Histology, Graduate School of Medical Sciences, Kumamoto University, Kumamoto 860-8556, Japan"},{"name":"Department of Neuroscience and Physiology, SUNY Upstate Medical University, Syracuse, NY 13210, USA"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"}]},{"given":"Mikiko","family":"Kudo","sequence":"additional","affiliation":[{"name":"Department of Stem Cell Biology, Graduate School of System Life Sciences, Kyushu University, Fukuoka 819-0395, Japan"},{"name":"School of Biomedical Sciences, Faculty of Medicine, The University of Queensland, Brisbane, QLD 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"}]},{"given":"Terumasa","family":"Umemoto","sequence":"additional","affiliation":[{"name":"International Research Center for Medical Sciences, Kumamoto University, Kumamoto 860-0811, Japan"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4748-2516","authenticated-orcid":false,"given":"Keisuke","family":"Yamashita","sequence":"additional","affiliation":[{"name":"Department of Stem Cell Biology, Graduate School of System Life Sciences, Kyushu University, Fukuoka 819-0395, Japan"},{"name":"Department of Stem Cell Biology, Faculty of Arts and Science, Kyushu University, Fukuoka 819-0395, Japan"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"resources","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"given":"Rie","family":"Kawano","sequence":"additional","affiliation":[{"name":"Department of Medical Oncology and Hematology, Oita University Faculty of Medicine, Oita 879-5593, Japan"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4564-9757","authenticated-orcid":false,"given":"Kunimasa","family":"Ohta","sequence":"additional","affiliation":[{"name":"Department of Stem Cell Biology, Graduate School of System Life Sciences, Kyushu University, Fukuoka 819-0395, Japan"},{"name":"Department of Stem Cell Biology, Faculty of Arts and Science, Kyushu University, Fukuoka 819-0395, Japan"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1152\/physrev.00016.2011","article-title":"New Fundamentals in Hemostasis","volume":"93","author":"Versteeg","year":"2013","journal-title":"Physiol. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1177\/0192623310389474","article-title":"Continuing Education Course #2: Current Understanding of Hemostasis","volume":"39","author":"Gale","year":"2010","journal-title":"Toxicol. Pathol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1152\/physrev.00067.2017","article-title":"Wound Healing: A Cellular Perspective","volume":"99","author":"Rodrigues","year":"2019","journal-title":"Physiol. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.matbio.2017.04.003","article-title":"The provisional matrix: Setting the stage for tissue repair outcomes","volume":"60\u201361","author":"Barker","year":"2017","journal-title":"Matrix Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1055\/s-0041-1731791","article-title":"Wound Healing: A Comprehensive Review","volume":"35","author":"Almadani","year":"2021","journal-title":"Semin. Plast. Surg."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1177\/0022034509359125","article-title":"Critical Review in Oral Biology & Medicine: Factors Affecting Wound Healing","volume":"89","author":"Guo","year":"2010","journal-title":"J. Dent. Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3691","DOI":"10.1182\/blood.V97.12.3691","article-title":"Wound-healing defects in mice lacking fibrinogen","volume":"97","author":"Drew","year":"2001","journal-title":"Blood"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2881","DOI":"10.1182\/blood.2020008951","article-title":"Plasminogen: An enigmatic zymogen","volume":"137","author":"Keragala","year":"2021","journal-title":"Blood"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1111\/j.1365-2141.2005.05444.x","article-title":"Molecular mechanisms of fibrinolysis","volume":"129","author":"Hajjar","year":"2005","journal-title":"Br. J. Haematol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"5879","DOI":"10.1182\/blood-2012-01-407825","article-title":"Plasminogen is a key proinflammatory regulator that accelerates the healing of acute and diabetic wounds","volume":"119","author":"Shen","year":"2012","journal-title":"Blood"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"3304","DOI":"10.1016\/j.jtha.2023.09.012","article-title":"The fibrinolysis renaissance","volume":"21","author":"Mutch","year":"2023","journal-title":"J. Thromb. Haemost."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"S98","DOI":"10.1111\/jth.12935","article-title":"Basic mechanisms and regulation of fibrinolysis","volume":"13","author":"Longstaff","year":"2015","journal-title":"J. Thromb. Haemost."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1200","DOI":"10.1111\/j.1538-7836.2011.04288.x","article-title":"Life-threatening hemorrhage and prolonged wound healing are remarkable phenotypes manifested by complete plasminogen activator inhibitor-1 deficiency in humans","volume":"9","author":"Iwaki","year":"2011","journal-title":"J. Thromb. Haemost."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1498","DOI":"10.1111\/jth.14160","article-title":"Hemorrhagic disorders of fibrinolysis: A clinical review","volume":"16","author":"Saes","year":"2018","journal-title":"J. Thromb. Haemost."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1002\/dvdy.20303","article-title":"Novel soluble molecule, Akhirin, is expressed in the embryonic chick eyes and exhibits heterophilic cell-adhesion activity","volume":"233","author":"Ahsan","year":"2005","journal-title":"Dev. Dyn."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"463","DOI":"10.1111\/dgd.12284","article-title":"Regulation of the neural niche by the soluble molecule Akhirin","volume":"58","author":"Acharjee","year":"2016","journal-title":"Dev. Growth Differ."},{"key":"ref_17","first-page":"494","article-title":"Akhirin regulates the proliferation and differentiation of neural stem cells in intact and injured mouse spinal cord","volume":"75","author":"M","year":"2014","journal-title":"Dev. Neurobiol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1111\/dgd.12646","article-title":"Akhirin regulates the proliferation and differentiation of neural stem cells\/progenitor cells at neurogenic niches in mouse brain","volume":"62","author":"Anam","year":"2020","journal-title":"Dev. Growth Differ."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Kudo, M., Ohkubo, T., Sugawara, T., Irie, T., Hatakeyama, J., Tamura, S., Shimamura, K., Wakayama, T., Matsuo, N., and Nakashima, K. (2026). Akhirin Functions as an Innate Immune Barrier to Preserve Neurogenic Niche Homeostasis During Mouse Brain Development. Cells, 15.","DOI":"10.3390\/cells15020151"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1487","DOI":"10.1111\/jth.14157","article-title":"Regulation of plasminogen activation on cell surfaces and fibrin","volume":"16","author":"Urano","year":"2018","journal-title":"J. Thromb. Haemost."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"30","DOI":"10.5493\/wjem.v2.i2.30","article-title":"Standardizing a simpler, more sensitive and accurate tail bleeding assay in mice","volume":"2","author":"Liu","year":"2012","journal-title":"World J. Exp. Med."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Kudo, M., and Ohta, K. (2021). Regulation of the Brain Neural Niche by Soluble Molecule Akhirin. J. Dev. Biol., 9.","DOI":"10.3390\/jdb9030029"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1941","DOI":"10.1111\/jth.14246","article-title":"Tissue factor at the crossroad of coagulation and cell signaling","volume":"16","author":"Zelaya","year":"2018","journal-title":"J. Thromb. Haemost."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"958","DOI":"10.1055\/s-0037-1615947","article-title":"A Cell-Based Model of Hemostasis","volume":"85","author":"Hoffman","year":"2001","journal-title":"Thromb. Haemost."},{"key":"ref_25","first-page":"1087","article-title":"Selective Cellular Expression of Tissue Factor in Human Tissues. Implications for Disorders of Hemostasis and Thrombosis","volume":"134","author":"Drake","year":"1989","journal-title":"Am. J. Pathol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"806","DOI":"10.1182\/blood-2009-12-259267","article-title":"Hematopoietic and nonhematopoietic cell tissue factor activates the coagulation cascade in endotoxemic mice","volume":"116","author":"Pawlinski","year":"2010","journal-title":"Blood"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1098\/rsif.2008.0327","article-title":"Fibrin gels and their clinical and bioengineering applications","volume":"6","author":"Janmey","year":"2009","journal-title":"J. R. Soc. Interface"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"932","DOI":"10.1111\/j.1538-7836.2006.01861.x","article-title":"Fibrin structure and wound healing","volume":"4","author":"Laurens","year":"2006","journal-title":"J. Thromb. Haemost."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1354","DOI":"10.1161\/01.ATV.20.5.1354","article-title":"Influence of Fibrin Network Conformation and Fibrin Fiber Diameter on Fibrinolysis Speed","volume":"20","author":"Collet","year":"2000","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1413","DOI":"10.1111\/j.1537-2995.2005.00546.x","article-title":"Transfusion Medicine\/Hemostasis Clinical Trials Network Paucity of studies to support that abnormal coagulation test results predict bleeding in the setting of invasive procedures: An evidence-based review","volume":"45","author":"Segal","year":"2005","journal-title":"Transfusion"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1038\/sj.thj.6200306","article-title":"Role of coagulation testing in predicting bleeding risk","volume":"4","author":"Chee","year":"2003","journal-title":"Hematol. J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1712","DOI":"10.1182\/blood-2012-09-306639","article-title":"Mechanisms of fibrin polymerization and clinical implications","volume":"121","author":"Weisel","year":"2013","journal-title":"Blood"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.blre.2014.09.003","article-title":"Fibrinolysis and the control of blood coagulation","volume":"29","author":"Chapin","year":"2015","journal-title":"Blood Rev."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1016\/j.tmrv.2014.09.006","article-title":"The Fibrinolytic System\u2014More Than Fibrinolysis?","volume":"29","author":"Draxler","year":"2015","journal-title":"Transfus. Med. Rev."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1038\/nrm2821","article-title":"Regulation of cell signalling by uPAR","volume":"11","author":"Smith","year":"2010","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"932","DOI":"10.1038\/nrm977","article-title":"uPAR: A versatile signalling orchestrator","volume":"3","author":"Blasi","year":"2002","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2686","DOI":"10.1038\/sj.emboj.7601173","article-title":"Plasminogen activation independent of uPA and tPA maintains wound healing in gene-deficient mice","volume":"25","author":"Lund","year":"2006","journal-title":"EMBO J."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"J\u00f6gi, A., R\u00f8n\u00f8, B., Lund, I.K., Nielsen, B.S., Ploug, M., H\u00f8yer-Hansen, G., R\u00f8mer, J., and Lund, L.R. (2010). Neutralisation of uPA with a Monoclonal Antibody Reduces Plasmin Formation and Delays Skin Wound Healing in tPA-Deficient Mice. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0012746"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"C721","DOI":"10.1152\/ajpcell.00269.2021","article-title":"Plasminogen activator receptor assemblies in cell signaling, innate immunity, and inflammation","volume":"321","author":"Gonias","year":"2021","journal-title":"Am. J. Physiol. Physiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"100081","DOI":"10.1016\/j.rpth.2023.100081","article-title":"Fibrinolysis: An illustrated review","volume":"7","author":"Risman","year":"2023","journal-title":"Res. Pract. Thromb. Haemost."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1026","DOI":"10.1126\/science.aam7928","article-title":"Inflammation and metabolism in tissue repair and regeneration","volume":"356","author":"Eming","year":"2017","journal-title":"Science"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1161\/ATVBAHA.117.309846","article-title":"Tissue Factor: An Essential Mediator of Hemostasis and Trigger of Thrombosis","volume":"38","author":"Grover","year":"2018","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.1161\/01.ATV.0000130465.23430.74","article-title":"Role of Tissue Factor in Hemostasis, Thrombosis, and Vascular Development","volume":"24","author":"Mackman","year":"2004","journal-title":"Arterioscler. Thromb. Vasc. Biol."}],"container-title":["Journal of Developmental Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2221-3759\/14\/3\/41\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T11:08:36Z","timestamp":1788779316000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2221-3759\/14\/3\/41"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,7]]},"references-count":43,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["jdb14030041"],"URL":"https:\/\/doi.org\/10.3390\/jdb14030041","relation":{},"ISSN":["2221-3759"],"issn-type":[{"value":"2221-3759","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,7]]}}}