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—microRNA-329-3p and microRNA-495-3p
Inhibition of the 14q32 microRNAs, miR-329-3p and miR-495-3p, improves post-ischemic neovascularization. Cold-inducible RNA-binding protein (<i>CIRBP</i>) facilitates maturation of these microRNAs. We hypothesized that <i>CIRBP</i> deficiency improves post-ischemic angiogenes...
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2021-11-01
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| Online Access: | https://www.mdpi.com/1422-0067/22/23/12678 |
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| author | Eveline A. C. Goossens Licheng Zhang Margreet R. de Vries J. Wouter Jukema Paul H. A. Quax A. Yaël Nossent |
| author_facet | Eveline A. C. Goossens Licheng Zhang Margreet R. de Vries J. Wouter Jukema Paul H. A. Quax A. Yaël Nossent |
| author_sort | Eveline A. C. Goossens |
| collection | DOAJ |
| description | Inhibition of the 14q32 microRNAs, miR-329-3p and miR-495-3p, improves post-ischemic neovascularization. Cold-inducible RNA-binding protein (<i>CIRBP</i>) facilitates maturation of these microRNAs. We hypothesized that <i>CIRBP</i> 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 <i>Cirp<sup>−/−</sup></i> 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 <i>Cirp<sup>−/−</sup></i> 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 <i>Cirp<sup>−/−</sup></i> mice by 40.6% and 36.2%. In HUVECs, <i>CIRBP</i> expression was upregulated under hypothermia, while miR-329-3p and miR-495-3p expression remained unaffected. siRNA-mediated <i>CIRBP</i> knockdown led to the downregulation of <i>CIRBP</i>-splice-variant-1 (<i>CIRBP-SV1</i>), <i>CIRBP</i> antisense long noncoding RNA (lncRNA-<i>CIRBP</i>-AS1), and miR-495-3p with no effects on the expression of <i>CIRBP-SV2-4</i> or miR-329-3p. siRNA-mediated <i>CIRBP</i> knockdown improved HUVEC migration and tube formation. SiRNA-mediated lncRNA-<i>CIRBP</i>-AS1 knockdown had similar long-term effects. After short incubation times, however, only <i>CIRBP</i> knockdown affected angiogenesis, indicating that the effects of lncRNA-<i>CIRBP</i>-AS1 knockdown were secondary to <i>CIRBP-SV1</i> downregulation. <i>CIRBP</i> 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. |
| first_indexed | 2024-03-10T04:53:00Z |
| format | Article |
| id | doaj.art-1f0b68cfae914a9ea7222f74c673513e |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T04:53:00Z |
| publishDate | 2021-11-01 |
| publisher | MDPI AG |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-1f0b68cfae914a9ea7222f74c673513e2023-11-23T02:26:20ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-11-0122231267810.3390/ijms222312678Cold-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—microRNA-329-3p and microRNA-495-3pEveline A. C. Goossens0Licheng Zhang1Margreet R. de Vries2J. Wouter Jukema3Paul H. A. Quax4A. Yaël Nossent5Department of Surgery, Leiden University Medical Center, 2333 ZA Leiden, The NetherlandsEinthoven Laboratory for Experimental Vascular Medicine, Leiden University Medical Center, 2333 ZA Leiden, The NetherlandsDepartment of Surgery, Leiden University Medical Center, 2333 ZA Leiden, The NetherlandsDepartment of Cardiology, Leiden University Medical Center, 2333 ZA Leiden, The NetherlandsDepartment of Surgery, Leiden University Medical Center, 2333 ZA Leiden, The NetherlandsDepartment of Surgery, Leiden University Medical Center, 2333 ZA Leiden, The NetherlandsInhibition of the 14q32 microRNAs, miR-329-3p and miR-495-3p, improves post-ischemic neovascularization. Cold-inducible RNA-binding protein (<i>CIRBP</i>) facilitates maturation of these microRNAs. We hypothesized that <i>CIRBP</i> 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 <i>Cirp<sup>−/−</sup></i> 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 <i>Cirp<sup>−/−</sup></i> 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 <i>Cirp<sup>−/−</sup></i> mice by 40.6% and 36.2%. In HUVECs, <i>CIRBP</i> expression was upregulated under hypothermia, while miR-329-3p and miR-495-3p expression remained unaffected. siRNA-mediated <i>CIRBP</i> knockdown led to the downregulation of <i>CIRBP</i>-splice-variant-1 (<i>CIRBP-SV1</i>), <i>CIRBP</i> antisense long noncoding RNA (lncRNA-<i>CIRBP</i>-AS1), and miR-495-3p with no effects on the expression of <i>CIRBP-SV2-4</i> or miR-329-3p. siRNA-mediated <i>CIRBP</i> knockdown improved HUVEC migration and tube formation. SiRNA-mediated lncRNA-<i>CIRBP</i>-AS1 knockdown had similar long-term effects. After short incubation times, however, only <i>CIRBP</i> knockdown affected angiogenesis, indicating that the effects of lncRNA-<i>CIRBP</i>-AS1 knockdown were secondary to <i>CIRBP-SV1</i> downregulation. <i>CIRBP</i> 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.https://www.mdpi.com/1422-0067/22/23/12678<i>CIRBP</i>14q32 microRNAsangiogenesisperipheral arterial diseaseHUVECs |
| spellingShingle | Eveline A. C. Goossens Licheng Zhang Margreet R. de Vries J. Wouter Jukema Paul H. A. Quax A. Yaël Nossent 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—microRNA-329-3p and microRNA-495-3p International Journal of Molecular Sciences <i>CIRBP</i> 14q32 microRNAs angiogenesis peripheral arterial disease HUVECs |
| 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—microRNA-329-3p and microRNA-495-3p |
| title_full | 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—microRNA-329-3p and microRNA-495-3p |
| title_fullStr | 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—microRNA-329-3p and microRNA-495-3p |
| title_full_unstemmed | 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—microRNA-329-3p and microRNA-495-3p |
| title_short | 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—microRNA-329-3p and microRNA-495-3p |
| title_sort | 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 microrna 329 3p and microrna 495 3p |
| topic | <i>CIRBP</i> 14q32 microRNAs angiogenesis peripheral arterial disease HUVECs |
| url | https://www.mdpi.com/1422-0067/22/23/12678 |
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