Single‐cell transcriptomic landscape of cardiac neural crest cell derivatives during development

Abstract

The migratory cardiac neural crest cells (CNCCs) contribute greatly to cardiovascular development. A thorough understanding of the cell lineages, developmental chronology, and transcriptomic states of CNCC derivatives during normal development is essential for deciphering the pathogenesis of CNCC-associated congenital anomalies. Here, we perform single-cell transcriptomic sequencing of 34,131 CNCC-derived cells in mouse hearts covering eight developmental stages between E10.5 and P7. We report the presence of CNCC-derived mural cells that comprise pericytes and microvascular smooth muscle cells (mVSMCs). Furthermore, we identify the transition from the CNCC-derived pericytes to mVSMCs and the key regulators over the transition. In addition, our data support that many CNCC derivatives had already committed or differentiated to a specific lineage when migrating into the heart. We explore the spatial distribution of some critical CNCC-derived subpopulations with single-molecule fluorescence in situ hybridization. Finally, we computationally reconstruct the differentiation path and regulatory dynamics of CNCC derivatives. Our study provides novel insights into the cell lineages, developmental chronology, and regulatory dynamics of CNCC derivatives during development.

Cardiac neural crest lineage diversity and underlying gene regulatory networks revealed by multimodal analysis

“…The copyright holder for this preprint this version posted June 24, 2022. ; https://doi.org/10.1101/2022.06.23.497419 doi: bioRxiv preprint respectively, as indicated by Chen and colleagues (see Discussion) 15 . Remarkably, intC0 and intC1 were connected by intC5 characterized by Sema3a expression, which is mainly composed of fC5 and fC6 (Fig.…”

“…But Tcf21 was highly expressed in intC4 as well as immature mesenchymal and valvular interstitial clusters intC0 and intC3. Unlike intC0, intC3 and Chen’s valvular interstitial cluster c11 15 , intC4 was also characterized by Ebf2 and allocated to tissues surrounding the great vessels in the 10x Visium platform, indicating a distinct cell population. Given that Ebf2 is a key TF regulating brown adipocyte development 28,29 , intC4 may be a mesenchymal cell population containing precursors of periaortic brown adipocytes, which were recently identified as of NC origin 30 .…”

“…Recently, Chen and colleagues reported single-cell transcriptomic landscape of cardiac NCCs, proposing the developmental path from NCCs migrating into the heart to mesenchymal and vascular smooth muscle lineages 15 . The NCC subpopulations and lineage relationship inferred in the present study are largely consistent with those reported by Chen et al, but there are some notable differences.…”

“…Given that Ebf2 is a key TF regulating brown adipocyte development 28,29 , intC4 may be a mesenchymal cell population containing precursors of periaortic brown adipocytes, which were recently identified as of NC origin 30 . Secondly, Chen et al designated the Penk -enriched cluster c9 as a cushion mesenchymal population involved in the formation of the aorticopulmonary septum 15 . By contrast, the present analysis found Penk expression broadly in smooth muscle(-like) and mesenchymal clusters, especially enriched in intC5, which bridged between the immature mesenchymal cluster intC0 and smooth muscle(-like) cluster intC1 (Supplementary Fig.…”

Cardiac neural crest lineage diversity and underlying gene regulatory networks revealed by multimodal analysis

“…IntC1 contained two subtypes as represented by mC7_0 (Myh11 +, Kcnj8 -) and mC6_1 (Myh11 + , Kcnj8 + ) (Supplementary Fig. 2l, 4i, l), which may correspond to smooth muscle cells (SMCs) of the great vessel and coronary artery SMCs, respectively, as indicated by Chen and colleagues (see Discussion) 15 . Remarkably, intC0 and intC1 were connected by intC5 characterized by Sema3a expression, which is mainly composed of fC5 and fC6 (Fig.…”