Prdm6 controls heart development by regulating neural crest cell differentiation and migration
Lingjuan Hong, Na Li, Victor Gasque, Sameet Mehta, Lupeng Ye, Yinyu Wu, Jinyu Li, Andreas Gewies, Jürgen Ruland, Karen K. Hirschi, Anne Eichmann, Caroline Hendry, David van Dijk, Arya Mani
Lingjuan Hong, Na Li, Victor Gasque, Sameet Mehta, Lupeng Ye, Yinyu Wu, Jinyu Li, Andreas Gewies, Jürgen Ruland, Karen K. Hirschi, Anne Eichmann, Caroline Hendry, David van Dijk, Arya Mani
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Research Article Cardiology Development

Prdm6 controls heart development by regulating neural crest cell differentiation and migration

Abstract

The molecular mechanisms that drive the acquisition of distinct neural crest cell (NCC) fates is still poorly understood. Here, we identified Prdm6 as an epigenetic modifier that temporally and spatially regulates the expression of NCC specifiers and determines the fate of a subset of migrating cardiac NCCs (CNCCs). Using transcriptomic analysis and genetic and fate mapping approaches in transgenic mice, we showed that disruption of Prdm6 was associated with impaired CNCC differentiation, delamination, and migration and led to patent ductus arteriosus (DA) and ventricular noncompaction. Bulk and single-cell RNA-Seq analyses of the DA and CNCCs identified Prdm6 as a regulator of a network of CNCC specification genes, including Wnt1, Tfap2b, and Sox9. Loss of Prdm6 in CNCCs diminished its expression in the pre-epithelial–mesenchymal transition (pre-EMT) cluster, resulting in the retention of NCCs in the dorsal neural tube. This defect was associated with diminished H4K20 monomethylation and G1-S progression and augmented Wnt1 transcript levels in pre-EMT and neural tube clusters, which we showed was the major driver of the impaired CNCC migration. Altogether, these findings revealed Prdm6 as a key regulator of CNCC differentiation and migration and identified Prdm6 and its regulated network as potential targets for the treatment of congenital heart diseases.

Authors

Lingjuan Hong, Na Li, Victor Gasque, Sameet Mehta, Lupeng Ye, Yinyu Wu, Jinyu Li, Andreas Gewies, Jürgen Ruland, Karen K. Hirschi, Anne Eichmann, Caroline Hendry, David van Dijk, Arya Mani

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Figure 3

Fate mapping of CNCCs in neural crest–specific Prdm6-deficient mice shows their reduced contribution to ductus arteriosus.

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Fate mapping of CNCCs in neural crest–specific Prdm6-deficient mice show...
(A) Representative cross-sections of DA of WT (top) and Prdm6fl/fl Wnt1-Cre2 ZsGreen1 (bottom) DA at E17.5 by confocal imaging; scale bar: 50 μm. (B) Percentage of neural crest–derived SMCs. (C) Magnified image demonstrating the colocalization of αSMA (gray) and ZsGreen1 (green). In contrast to controls, the DA wall was thinner and only a fraction of SMCs of Prdm6fl/fl Wnt1-Cre2 ZsGreen1 were NCC derived. In addition, the NCC-derived SMCs of Prdm6fl/fl Wnt1-Cre2 ZsGreen1 have an irregular shape (outlined by straight yellow lines) and have infiltrated and disrupted the endothelial layer, stained with CD31 antibody (outlined by dotted yellow lines); scale bar: 20 μm. (D) The confocal images of representative cross-sections of DA of control (top row) and Prdm6fl/fl Wnt1-Cre2 ZsGreen1 (bottom row) mice at E17.5 stained for Ki67 (red) and αSMA (gray) show the localization of ZsGreen1 (green) and Ki67 (white arrowheads) and Ki67 in ZsGreen1-negative SMCs, indicated by yellow arrowheads. Scale bar: 50 μm. (E) Percentage of Ki67-positive cells in NCC-derived SMCs in WT mice and NCC-derived and non-NCC-derived SMCs in Prdm6fl/fl Wnt1-Cre2 ZsGreen1 (KO) DA. All SMCs in WT mice were NCC derived. There was no significant difference in the percentage of Ki67-positive NCC-derived SMCs between WT and KO mice. The image intensities were quantified by ImageJ, and the thresholds for positive color detection were kept constant between different images. Each dot represents a biological replicate. The comparison between different groups was done by a 2-tailed unpaired t test, and data are shown as mean ± SEM. The comparisons between multiple groups (E) were done by 1-way ANOVA. A Mann-Whitney test was conducted for non-normally distributed data. ****P < 0.0001. All controls were the corresponding littermates (n = 5–10 per group).