{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T16:20:11Z","timestamp":1787674811883,"version":"build-2784847793"},"reference-count":24,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T00:00:00Z","timestamp":1786060800000},"content-version":"vor","delay-in-days":6,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"},{"start":{"date-parts":[[2026,8,1]],"date-time":"2026-08-01T00:00:00Z","timestamp":1785542400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32161143021"],"award-info":[{"award-number":["32161143021"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81271410"],"award-info":[{"award-number":["81271410"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Anim Models and Exp Med"],"published-print":{"date-parts":[[2026,8]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Venous malformations (VMs) are congenital vascular anomalies characterized by abnormal vascular proliferation, with limb VMs often leading to functional impairment and physical discomfort. However, the cellular heterogeneity and underlying molecular mechanisms driving pathological proliferation in limb VMs remain incompletely elucidated.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>In this study, we collected 10 tissue samples including specimens from 5 limb VM patients and 5 normal control tissues and performed single\u2010nucleus RNA sequencing (snRNA\u2010seq) to comprehensively map the cellular landscape of VMs. We first identified distinct cell subpopulations covering vascular endothelial cells, vascular smooth muscle cells, and immune cells, and compared gene expression levels between VM and normal tissues. Afterward, we conducted weighted gene co\u2010expression network analysis (hdWGCNA) and constructed the protein\u2013protein interaction (PPI) network to screen critical proliferation\u2010related genes in VMs. We further analyzed the signaling pathways associated with these candidate genes and carried out subsequent functional validation experiments to explore the biological role of core gene TEK in proliferative vascular endothelial cells (PVECs). Besides, we also analyzed the characteristic pathways of the PVEC subpopulation to clarify its proliferation\u2010related molecular features.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>We found obvious expression differences of genes in various cell subpopulations between VM and normal tissues. Three genes, namely tyrosine protein kinase receptor (TEK), Fms\u2010like tyrosine kinase 1 (FLT1), and EGF\u2010like domain multiple 7 (EGFL7), were identified as key proliferation\u2010related genes with significant upregulation in VM lesions, and these three genes were closely associated with the activation of PI3K\/AKT\/mTOR, IL6\/JAK\/STAT3, and TNF\u2010\u03b1\/NF\u2010\u03baB pathways. Functional experimental results showed that TEK knockdown could significantly inhibit proliferative vascular endothelial cell (PVEC) proliferation and promote cell apoptosis, and reverse the abnormal activation of inflammatory pathways. As a vital pathogenic cell subpopulation, PVECs facilitated abnormal vascular proliferation through activating pathways including the G2\/M checkpoint and E2F targets.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>Collectively, our study systematically elucidated the cellular heterogeneity framework and proliferation mechanisms of limb VMs, identifying TEK, FLT1, and EGFL7 as key regulators of pathological proliferation. These findings provide new insights into the pathogenesis of VMs and lay a foundation for developing precise therapeutic strategies targeting proliferation\u2010related pathways.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/ame2.70256","type":"journal-article","created":{"date-parts":[[2026,8,8]],"date-time":"2026-08-08T05:28:41Z","timestamp":1786166921000},"page":"1567-1586","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["<scp>SnRNA<\/scp>\n                    \u2010seq reveals cellular heterogeneity and proliferation mechanisms in limb venous malformations"],"prefix":"10.1002","volume":"9","author":[{"given":"Junjie","family":"Lin","sequence":"first","affiliation":[{"name":"Department of Hemangioma Surgery The Third Affiliated Hospital of Zhengzhou University  Zhengzhou Henan China"},{"name":"Key Discipline of Hemangioma and Vascular Malformation Medicine in Henan Province  Zhengzhou Henan China"},{"name":"Key Laboratory of Hemangioma and Vascular Malformation Medicine in Henan Province  Zhengzhou Henan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tingting","family":"Liu","sequence":"additional","affiliation":[{"name":"Key Discipline of Hemangioma and Vascular Malformation Medicine in Henan Province  Zhengzhou Henan China"},{"name":"Key Laboratory of Hemangioma and Vascular Malformation Medicine in Henan Province  Zhengzhou Henan China"},{"name":"Institute for Brain Sciences Research, School of Life Sciences Henan University  Kaifeng Henan China"},{"name":"Institute for Sports and Brain Health, School of Physical Education Henan University  Kaifeng Henan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bin","family":"Fang","sequence":"additional","affiliation":[{"name":"Department of Hemangioma Surgery The Third Affiliated Hospital of Zhengzhou University  Zhengzhou Henan China"},{"name":"Key Discipline of Hemangioma and Vascular Malformation Medicine in Henan Province  Zhengzhou Henan China"},{"name":"Key Laboratory of Hemangioma and Vascular Malformation Medicine in Henan Province  Zhengzhou Henan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaojuan","family":"Feng","sequence":"additional","affiliation":[{"name":"Department of Hemangioma Surgery The Third Affiliated Hospital of Zhengzhou University  Zhengzhou Henan China"},{"name":"Key Discipline of Hemangioma and Vascular Malformation Medicine in Henan Province  Zhengzhou Henan China"},{"name":"Key Laboratory of Hemangioma and Vascular Malformation Medicine in Henan Province  Zhengzhou Henan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenting","family":"Jiao","sequence":"additional","affiliation":[{"name":"Department of Hemangioma Surgery The Third Affiliated Hospital of Zhengzhou University  Zhengzhou Henan China"},{"name":"Key Discipline of Hemangioma and Vascular Malformation Medicine in Henan 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