The Department of Cleft Lip and Palate of Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Medical Cosmetic Center of Dermatology Hospital of Southern Medical University, Guangdong Provincial Dermatology Hospital, Guangzhou, China.
J Cell Mol Med. 2024 Oct;28(19):e70128. doi: 10.1111/jcmm.70128.
Cleft lip and/or primary palate (CL/P) represent a prevalent congenital malformation, the aetiology of which is highly intricate. Although it is generally accepted that the condition arises from failed fusion between the upper lip and primary palate, the precise mechanism underlying this fusion process remains enigmatic. In this study, we utilized transposase-accessible chromatin sequencing (scATAC-seq) and single-cell RNA sequencing (scRNA-seq) to interrogate lambdoidal junction tissue derived from C57BL/6J mouse embryos at critical stages of embryogenesis (10.5, 11.5 and 12.5 embryonic days). We successfully identified distinct subgroups of mesenchymal and ectodermal cells involved in the fusion process and characterized their unique transcriptional profiles. Furthermore, we conducted cell differentiation trajectory analysis, revealing a dynamic repertoire of genes that are sequentially activated or repressed during pseudotime, facilitating the transition of relevant cell types. Additionally, we employed scATAC data to identify key genes associated with the fusion process and demonstrated differential chromatin accessibility across major cell types. Finally, we constructed a dynamic intercellular communication network and predicted upstream transcriptional regulators of critical genes involved in important signalling pathways. Our findings provide a valuable resource for future studies on upper lip and primary palate development, as well as congenital defects.
唇裂和/或腭裂（CL/P）是一种常见的先天性畸形，其病因非常复杂。虽然人们普遍认为该疾病是由于上唇和初级腭未能融合而引起的，但这种融合过程的确切机制仍不清楚。在这项研究中，我们利用转座酶可及染色质测序（scATAC-seq）和单细胞 RNA 测序（scRNA-seq）技术，研究了源自 C57BL/6J 小鼠胚胎在胚胎发生关键阶段（10.5、11.5 和 12.5 天）的人字缝组织。我们成功地鉴定了参与融合过程的间质和外胚层细胞的不同亚群，并对其独特的转录谱进行了表征。此外，我们进行了细胞分化轨迹分析，揭示了在伪时间过程中顺序激活或抑制的一系列基因，促进了相关细胞类型的转变。此外，我们还利用 scATAC 数据来识别与融合过程相关的关键基因，并证明了主要细胞类型之间的染色质可及性存在差异。最后，我们构建了一个动态的细胞间通讯网络，并预测了重要信号通路中关键基因的上游转录调节因子。我们的研究结果为上唇和初级腭发育以及先天性缺陷的进一步研究提供了有价值的资源。
