Elevated microRNA-187 causes cardiac endothelial dysplasia to promote congenital heart disease through inhibition of NIPBL
Chao Li, … , Jianfeng Shen, Hongyan Wang
Chao Li, … , Jianfeng Shen, Hongyan Wang
Published November 25, 2024
Citation Information: J Clin Invest. 2025;135(1):e178355. https://doi.org/10.1172/JCI178355.
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Research Article Cardiology Development

Elevated microRNA-187 causes cardiac endothelial dysplasia to promote congenital heart disease through inhibition of NIPBL

Abstract

Cardiac endothelial cells are essential for heart development, and disruption of this process can lead to congenital heart disease (CHD). However, how microRNAs influence cardiac endothelial cells in CHD remains unclear. This study identified elevated microRNA-187 (miR-187) expression in embryonic heart endothelial cells from CHD fetuses. Using a conditional knockin model, we showed that increased miR-187 levels in embryonic endothelial cells induce CHD in homozygous fetal mice, closely mirroring human CHD. Mechanistically, miR-187 targets NIPBL, which is responsible for recruiting the cohesin complex and facilitating chromatin accessibility. Consequently, the endothelial cell–specific upregulation of miR-187 inhibited NIPBL, leading to reduced chromatin accessibility and impaired gene expression, which hindered endothelial cell development and ultimately caused heart septal defects and reduced heart size both in vitro and in vivo. Importantly, exogenous miR-187 expression in human cardiac organoids mimicked developmental defects in the cardiac endothelial cells, and this was reversible by NIPBL replenishment. Our findings establish the miR-187/NIPBL axis as a potent regulator that inhibits cardiac endothelial cell development by attenuating the transcription of numerous endothelial genes, with our mouse and human cardiac organoid models effectively replicating severe defects from minor perturbations. This discovery suggests that targeting the miR-187/NIPBL pathway could offer a promising therapeutic approach for CHD.

Authors

Chao Li, Zizheng Tan, Hongdou Li, Xiaoying Yao, Chuyue Peng, Yue Qi, Bo Wu, Tongjin Zhao, Chentao Li, Jianfeng Shen, Hongyan Wang

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Figure 4

MiR-187 targets NIPBL and disturbs endothelial development.

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MiR-187 targets NIPBL and disturbs endothelial development. (A) Schemati...
(A) Schematic illustration of the screening approach for target genes of miR-187 using TargetScan prediction, MGI database, and RT-qPCR verification. (B) Cluster analysis of RT-qPCR results showing expression levels of candidate target genes of miR-187 in hESC-ECs transfected with miR-187 mimic. (C) Schematic illustration of luciferase reporters containing WT and mutant miR-187 binding sites in the NIPBL 3′-UTR. (D) Luciferase assays of hESC-ECs or HEK293T cells cotransfected with miR-187 or scramble control and luciferase reporter plasmids containing WT or mutant NIPBL 3′-UTR. (E) The human NIPBL 3′-UTR pulled down by biotin–miR-187 or biotin-scramble control was quantified by RT-qPCR in hESC-ECs. SMAD7–3′-UTR and GAPDH–3′-UTR serve as positive and negative controls, respectively. (F and G) RT-qPCR and Western blotting was used to analyze mRNA (F) and protein (G, WB) levels of NIPBL in hESC-ECs transfected with miR-187 or scrambled control, with grayscale analysis used to quantify the NIPBL protein (G, statistical analysis). (H) RT-qPCR analysis of mRNA levels of NIPBL in RVs of aborted fetuses with TOF and control fetuses (n = 5). (I and J) mRNA (I) and protein (J) level of NIPBL in whole hearts, cardiac endothelial cells, and cardiomyocytes of P0 neonatal mice of the indicated genotypes (n = 6). (K and L) FACS quantification of CD31-positive cells in hESC-EC infection with sh-NIPBL-1, -2, -3, sh-scramble (K), pri-miR-187, pri-miR-187+NIPBL, or scramble control by lentivirus (n = 4) (L). (M and N) RT-qPCR analyses of expression levels of various markers for early endothelial (M) and mature endothelial cells (N) during differentiation from hESCs to endothelial cells (n = 4). GAPDH or H3 was used as an internal control. Data are shown as means ± SD. ns, P > 0.05; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Significance was determined by 1-way ANOVA (D, E, and I–L), 2-way ANOVA (M and N), and 2-tailed t test (F–H).