miR-96 and miR-183 differentially regulate neonatal and adult postinfarct neovascularization
Raphael F.P. Castellan, … , Andrea Caporali, Marco Meloni
Raphael F.P. Castellan, … , Andrea Caporali, Marco Meloni
Published June 16, 2020
Citation Information: JCI Insight. 2020;5(14):e134888. https://doi.org/10.1172/jci.insight.134888.
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Research Article Angiogenesis Vascular biology

miR-96 and miR-183 differentially regulate neonatal and adult postinfarct neovascularization

Abstract

Following myocardial infarction (MI), the adult heart has minimal regenerative potential. Conversely, the neonatal heart can undergo extensive regeneration, and neovascularization capacity was hypothesized to contribute to this difference. Here, we demonstrate the higher angiogenic potential of neonatal compared with adult mouse cardiac endothelial cells (MCECs) in vitro and use this difference to identify candidate microRNAs (miRs) regulating cardiac angiogenesis after MI. miR expression profiling revealed miR-96 and miR-183 upregulation in adult compared with neonatal MCECs. Their overexpression decreased the angiogenic potential of neonatal MCECs in vitro and prevented scar resolution and neovascularization in neonatal mice after MI. Inversely, their inhibition improved the angiogenic potential of adult MCECs, and miR-96/miR-183–KO mice had increased peri-infarct neovascularization. In silico analyses identified anillin (ANLN) as a direct target of miR-96 and miR-183. In agreement, Anln expression declined following their overexpression and increased after their inhibition in vitro. Moreover, ANLN expression inversely correlated with miR-96 expression and age in cardiac ECs of cardiovascular patients. In vivo, ANLN+ vessels were enriched in the peri-infarct area of miR-96/miR-183–KO mice. These findings identify miR-96 and miR-183 as regulators of neovascularization following MI and miR-regulated genes, such as anillin, as potential therapeutic targets for cardiovascular disease.

Authors

Raphael F.P. Castellan, Milena Vitiello, Martina Vidmar, Steven Johnstone, Dominga Iacobazzi, David Mellis, Benjamin Cathcart, Adrian Thomson, Christiana Ruhrberg, Massimo Caputo, David E. Newby, Gillian A. Gray, Andrew H. Baker, Andrea Caporali, Marco Meloni

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Figure 1

Morphological and functional differences between neonatal and adult MCECs.

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Morphological and functional differences between neonatal and adult MCEC...
(A) Graphs showing increased cell-cell contacts and cell-surface adhesions of neonatal mouse cardiac endothelial cells (MCECs) compared with adult MCECs. (B) Representative impedance curve and quantification showing accelerated migration of neonatal MCECs (red) compared with adult MCECs (black). (C). Representative microphotographs and bar graph showing the higher percentage of EdU-proliferating cells in neonatal versus adult MCECs after 24 hours in culture. EdU+ cells are shown in red and indicated by arrows, DAPI+ nuclei are shown in blue. (D) Matrigel experiment with MCECs seeded at high density (2 × 104) showing that adult MCECs are able to form tube-like structures at 6 hours in culture properly and demonstrating that the length of tube-like structures formed by the same number of neonatal MCECs is higher. (E) Representative microphotographs and bar graph showing that, whereas 1 × 104 neonatal MCECs form proper tube-like structures at 24 hours, adult MCECs barely form tube-like structure at the same time point. Scale bar: 50 μm. n = 3 (B); n = 4/group (A, C, D, and E). *P < 0.05, **P < 0.01 vs. adult MCECs (Student’s t test). Data are shown as mean ± SEM.