{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T02:49:10Z","timestamp":1790650150609,"version":"4.1.0"},"reference-count":39,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2021,11,24]],"date-time":"2021-11-24T00:00:00Z","timestamp":1637712000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002428","name":"FWF Austrian Science Fund","doi-asserted-by":"publisher","award":["M-2578-B30"],"award-info":[{"award-number":["M-2578-B30"]}],"id":[{"id":"10.13039\/501100002428","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Inhibition of the 14q32 microRNAs, miR-329-3p and miR-495-3p, improves post-ischemic neovascularization. Cold-inducible RNA-binding protein (CIRBP) facilitates maturation of these microRNAs. We hypothesized that CIRBP deficiency improves post-ischemic angiogenesis via downregulation of 14q32 microRNA expression. We investigated these regulatory mechanisms both in vitro and in vivo. We induced hindlimb ischemia in Cirp\u2212\/\u2212 and C57Bl\/6-J mice, monitored blood flow recovery with laser Doppler perfusion imaging, and assessed neovascularization via immunohistochemistry. Post-ischemic angiogenesis was enhanced in Cirp\u2212\/\u2212 mice by 34.3% with no effects on arteriogenesis. In vivo at day 7, miR-329-3p and miR-495-3p expression were downregulated in Cirp\u2212\/\u2212 mice by 40.6% and 36.2%. In HUVECs, CIRBP expression was upregulated under hypothermia, while miR-329-3p and miR-495-3p expression remained unaffected. siRNA-mediated CIRBP knockdown led to the downregulation of CIRBP-splice-variant-1 (CIRBP-SV1), CIRBP antisense long noncoding RNA (lncRNA-CIRBP-AS1), and miR-495-3p with no effects on the expression of CIRBP-SV2-4 or miR-329-3p. siRNA-mediated CIRBP knockdown improved HUVEC migration and tube formation. SiRNA-mediated lncRNA-CIRBP-AS1 knockdown had similar long-term effects. After short incubation times, however, only CIRBP knockdown affected angiogenesis, indicating that the effects of lncRNA-CIRBP-AS1 knockdown were secondary to CIRBP-SV1 downregulation. CIRBP is a negative regulator of angiogenesis in vitro and in vivo and acts, at least in part, through the regulation of miR-329-3p and miR-495-3p.<\/jats:p>","DOI":"10.3390\/ijms222312678","type":"journal-article","created":{"date-parts":[[2021,11,30]],"date-time":"2021-11-30T04:48:37Z","timestamp":1638247717000},"page":"12678","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Cold-Inducible RNA-Binding Protein but Not Its Antisense lncRNA Is a Direct Negative Regulator of Angiogenesis In Vitro and In Vivo via Regulation of the 14q32 angiomiRs\u2014microRNA-329-3p and microRNA-495-3p"],"prefix":"10.3390","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5080-9639","authenticated-orcid":false,"given":"Eveline A. C.","family":"Goossens","sequence":"first","affiliation":[{"name":"Department of Surgery, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands"},{"name":"Einthoven Laboratory for Experimental Vascular Medicine, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Licheng","family":"Zhang","sequence":"additional","affiliation":[{"name":"Einthoven Laboratory for Experimental Vascular Medicine, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands"},{"name":"Department of Cardiology, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3648-9130","authenticated-orcid":false,"given":"Margreet R.","family":"de Vries","sequence":"additional","affiliation":[{"name":"Department of Surgery, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands"},{"name":"Einthoven Laboratory for Experimental Vascular Medicine, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3246-8359","authenticated-orcid":false,"given":"J. Wouter","family":"Jukema","sequence":"additional","affiliation":[{"name":"Department of Cardiology, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands"},{"name":"Netherlands Heart Institute, 3511 EP Utrecht, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6853-5760","authenticated-orcid":false,"given":"Paul H. A.","family":"Quax","sequence":"additional","affiliation":[{"name":"Department of Surgery, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands"},{"name":"Einthoven Laboratory for Experimental Vascular Medicine, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2155-4286","authenticated-orcid":false,"given":"A. Ya\u00ebl","family":"Nossent","sequence":"additional","affiliation":[{"name":"Department of Surgery, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands"},{"name":"Einthoven Laboratory for Experimental Vascular Medicine, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands"},{"name":"Department of Laboratory Medicine, Medical University of Vienna, 1090 Wien, Austria"},{"name":"Department of Internal Medicine II, Medical University of Vienna, 1090 Wien, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,11,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1540","DOI":"10.1161\/CIRCRESAHA.114.303517","article-title":"Lower extremity manifestations of peripheral artery disease: The pathophysiologic and functional implications of leg ischemia","volume":"116","author":"McDermott","year":"2015","journal-title":"Circ. Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1614","DOI":"10.1161\/CIRCRESAHA.116.303504","article-title":"Surgical intervention for peripheral arterial disease","volume":"116","author":"Vartanian","year":"2015","journal-title":"Circ. Res."},{"key":"ref_3","first-page":"10595","article-title":"Angioplasty versus bypass surgery in patients with critical limb ischemia-a meta-analysis","volume":"8","author":"Fu","year":"2015","journal-title":"Int. J. Clin. Exp. Med."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Parvar, S.L., Ngo, L., Dawson, J., Nicholls, S.J., Fitridge, R., Psaltis, P.J., and Ranasinghe, I. (2021). Long-term outcomes following endovascular and surgical revascularization for peripheral artery disease: A propensity score-matched analysis. Eur. Heart J., ehab116.","DOI":"10.1093\/eurheartj\/ehab116"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Kubler, M., Beck, S., Fischer, S., G\u00f6tz, P., Kumaraswami, K., Ishikawa-Ankerhold, H., Lasch, M., and Deindl, E. (2021). Absence of Cold-Inducible RNA-Binding Protein (CIRP) Promotes Angiogenesis and Regeneration of Ischemic Tissue by Inducing M2-Like Macrophage Polarization. Biomedicines, 9.","DOI":"10.3390\/biomedicines9040395"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Kubler, M., Beck, S., Peffenk\u00f6ver, L.L., G\u00f6tz, P., Ishikawa-Ankerhold, H., Preissner, K.T., Fischer, S., Lasch, M., and Deindl, E. (2021). The Absence of Extracellular Cold-Inducible RNA-Binding Protein (eCIRP) Promotes Pro-Angiogenic Microenvironmental Conditions and Angiogenesis in Muscle Tissue Ischemia. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22179484"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1016\/j.brainres.2010.05.029","article-title":"Effects of hypothermia and cerebral ischemia on cold-inducible RNA-binding protein mRNA expression in rat brain","volume":"134","author":"Liu","year":"2010","journal-title":"Brain Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1785","DOI":"10.1242\/jcs.01026","article-title":"Oxygen-regulated expression of the RNA-binding proteins RBM3 and CIRP by a HIF-1-independent mechanism","volume":"117","author":"Wellmann","year":"2004","journal-title":"J. Cell Sci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2164","DOI":"10.1002\/ijc.30833","article-title":"The role of cold-inducible RNA binding protein in cell stress response","volume":"141","author":"Liao","year":"2017","journal-title":"Int. J. Cancer"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1164","DOI":"10.1261\/rna.1179109","article-title":"Cold-inducible RNA binding protein (CIRP) expression is modulated by alternative mRNAs","volume":"15","author":"Smales","year":"2009","journal-title":"RNA"},{"key":"ref_11","first-page":"43","article-title":"A new generation of proto-oncogenes: Cold-inducible RNA binding proteins","volume":"1805","author":"Lleonart","year":"2010","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_12","first-page":"243","article-title":"Cold shock response in mammalian cells","volume":"1","author":"Fujita","year":"1999","journal-title":"J. Mol. Microbiol. Biotechnol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"899","DOI":"10.1083\/jcb.137.4.899","article-title":"A glycine-rich RNA-binding protein mediating cold-inducible suppression of mammalian cell growth","volume":"137","author":"Nishiyama","year":"1997","journal-title":"J. Cell Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/S0378-1119(97)00530-1","article-title":"Cloning and characterization of human CIRP (cold-inducible RNA-binding protein) cDNA and chromosomal assignment of the gene","volume":"204","author":"Nishiyama","year":"1997","journal-title":"Gene"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3839","DOI":"10.1007\/s00018-016-2253-7","article-title":"Cold-inducible proteins CIRP and RBM3, a unique couple with activities far beyond the cold","volume":"73","author":"Zhu","year":"2016","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"FSO246","DOI":"10.4155\/fsoa-2017-0077","article-title":"Recent progress in the research of cold-inducible RNA-binding protein","volume":"3","author":"Zhong","year":"2017","journal-title":"Future Sci. OA"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"153","DOI":"10.2220\/biomedres.40.153","article-title":"Hypothermia induces changes in the alternative splicing pattern of cold-inducible RNA-binding protein transcripts in a non-hibernator, the mouse","volume":"40","author":"Horii","year":"2019","journal-title":"Biomed Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"880","DOI":"10.1038\/nrg3594","article-title":"Gene regulation by antisense transcription","volume":"14","author":"Pelechano","year":"2013","journal-title":"Nat. Rev. Genet."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Gomes, C.P., N\u00f3brega-Pereira, S., Silva, A.B.D., Rebelo, K., Alves-Vale, C., Marinho, S.P., Carvalho, T., Dias, S., and De Jesus, B.B. (2019). An antisense transcript mediates MALAT1 response in human breast cancer. BMC Cancer, 19.","DOI":"10.1186\/s12885-019-5962-0"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2898","DOI":"10.1093\/nar\/gkw047","article-title":"Natural antisense RNA promotes 3\u2019 end processing and maturation of MALAT1 lncRNA","volume":"44","author":"Zong","year":"2016","journal-title":"Nucleic Acids Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"696","DOI":"10.1161\/CIRCRESAHA.114.304747","article-title":"Inhibition of 14q32 MicroRNAs miR-329, miR-487b, miR-494, and miR-495 increases neovascularization and blood flow recovery after ischemia","volume":"115","author":"Welten","year":"2014","journal-title":"Circ. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.atherosclerosis.2017.04.011","article-title":"Inhibition of 14q32 microRNA miR-495 reduces lesion formation, intimal hyperplasia and plasma cholesterol levels in experimental restenosis","volume":"261","author":"Welten","year":"2017","journal-title":"Atherosclerosis"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"841","DOI":"10.1097\/SLA.0000000000001466","article-title":"Inhibition of MicroRNA-494 Reduces Carotid Artery Atherosclerotic Lesion Development and Increases Plaque Stability","volume":"262","author":"Wezel","year":"2015","journal-title":"Ann. Surg."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1093\/cvr\/cvw039","article-title":"The multifactorial nature of microRNAs in vascular remodelling","volume":"110","author":"Welten","year":"2016","journal-title":"Cardiovasc. Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1016\/j.omtn.2018.11.017","article-title":"Posttranscriptional Regulation of 14q32 MicroRNAs by the CIRBP and HADHB during Vascular Regeneration after Ischemia","volume":"14","author":"Welten","year":"2019","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1042\/CS20150435","article-title":"Evaluation of the clinical relevance and limitations of current pre-clinical models of peripheral artery disease","volume":"130","author":"Krishna","year":"2016","journal-title":"Clin. Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1989","DOI":"10.1161\/ATVBAHA.108.175463","article-title":"Angiopoietin-2 stimulates blood flow recovery after femoral artery occlusion by inducing inflammation and arteriogenesis","volume":"28","author":"Tressel","year":"2008","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"406","DOI":"10.1055\/s-0038-1676342","article-title":"On the Cutting Edge: Wound Care for the Endovascular Specialist","volume":"35","author":"Olivieri","year":"2018","journal-title":"Semin. Intervent. Radiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"423","DOI":"10.3892\/ijmm.2016.2451","article-title":"A deficiency in cold-inducible RNA-binding protein accelerates the inflammation phase and improves wound healing","volume":"37","author":"Idrovo","year":"2016","journal-title":"Int. J. Mol. Med."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1489","DOI":"10.1038\/nm.3368","article-title":"Cold-inducible RNA-binding protein (CIRP) triggers inflammatory responses in hemorrhagic shock and sepsis","volume":"19","author":"Qiang","year":"2013","journal-title":"Nat. Med."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.yjmcc.2010.08.006","article-title":"Arteriogenesis requires toll-like receptor 2 and 4 expression in bone-marrow derived cells","volume":"50","author":"Hoefer","year":"2011","journal-title":"J. Mol. Cell. Cardiol."},{"key":"ref_32","first-page":"289","article-title":"Decreased expression of cold-inducible RNA-binding protein (CIRP) in male germ cells at elevated temperature","volume":"152","author":"Nishiyama","year":"1998","journal-title":"Am. J. Pathol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1080\/23328940.2019.1614851","article-title":"Serum from young, sedentary adults who underwent passive heat therapy improves endothelial cell angiogenesis via improved nitric oxide bioavailability","volume":"6","author":"Brunt","year":"2019","journal-title":"Temperature"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"R392","DOI":"10.1152\/ajpregu.00147.2016","article-title":"Thermotherapy reduces blood pressure and circulating endothelin-1 concentration and enhances leg blood flow in patients with symptomatic peripheral artery disease","volume":"311","author":"Neff","year":"2016","journal-title":"Am. J. Physiol. Regul. Integr. Comp. Physiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"10885","DOI":"10.1073\/pnas.1121524109","article-title":"Cold-inducible RNA-binding protein (Cirp) interacts with Dyrk1b\/Mirk and promotes proliferation of immature male germ cells in mice","volume":"109","author":"Masuda","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_36","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_37","doi-asserted-by":"crossref","first-page":"1902","DOI":"10.1161\/ATVBAHA.113.301579","article-title":"Lysine acetyltransferase PCAF is a key regulator of arteriogenesis","volume":"33","author":"Bastiaansen","year":"2013","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"e005281","DOI":"10.1161\/JAHA.116.005281","article-title":"CCR7-CCL19\/CCL21 Axis is Essential for Effective Arteriogenesis in a Murine Model of Hindlimb Ischemia","volume":"6","author":"Nossent","year":"2017","journal-title":"J. Am. Heart Assoc."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Parma, L., Peters, H.A., Johansson, M.E., Guti\u00e9rrez, S., Meijerink, H., De Kimpe, S., De Vries, M.R., and Quax, P. (2020). Bis(maltolato)oxovanadium(IV) Induces Angiogenesis via Phosphorylation of VEGFR2. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21134643"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/23\/12678\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:35:00Z","timestamp":1760168100000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/23\/12678"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,11,24]]},"references-count":39,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["ijms222312678"],"URL":"https:\/\/doi.org\/10.3390\/ijms222312678","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,11,24]]}}}