This study aimed to gain insights into the understanding of placenta accreta spectrum disorder by using single-cell and spatially resolved transcriptomics to characterize cellular heterogeneity at the maternal-fetal interface in placenta accreta spectrum disorders.
To assess cellular heterogeneity and the function of cell types, single-cell RNA sequencing and spatially resolved transcriptomics were used. A total of 12 placentas were included, 6 placentas with placenta accreta spectrum disorder and 6 controls. For each placenta with placenta accreta spectrum disorder, multiple biopsies were taken at the following sites: placenta accreta spectrum adherent and nonadherent sites in the same placenta. Of note, 2 platforms were used to generate libraries: the 10× Chromium and NanoString GeoMX Digital Spatial Profiler for single-cell and spatially resolved transcriptomes, respectively. Differential gene expression analysis was performed using a suite of bioinformatic tools (Seurat and GeoMxTools R packages). Correction for multiple testing was performed using Clipper. In situ hybridization was performed with RNAscope, and immunohistochemistry was used to assess protein expression.
In creating a placenta accreta cell atlas, there were dramatic difference in the transcriptional profile by site of biopsy between placenta accreta spectrum and controls. Most of the differences were noted at the site of adherence; however, differences existed within the placenta between the adherent and nonadherent site of the same placenta in placenta accreta. Among all cell types, the endothelial-stromal populations exhibited the greatest difference in gene expression, driven by changes in collagen genes, namely collagen type III alpha 1 chain (COL3A1), growth factors, epidermal growth factor-like protein 6 (EGFL6), and hepatocyte growth factor (HGF), and angiogenesis-related genes, namely delta-like noncanonical Notch ligand 1 (DLK1) and platelet endothelial cell adhesion molecule-1 (PECAM1). Intraplacental tropism (adherent versus non-adherent sites in the same placenta) was driven by differences in endothelial-stromal cells with notable differences in bone morphogenic protein 5 (BMP5) and osteopontin (SPP1) in the adherent vs nonadherent site of placenta accreta spectrum.
Placenta accreta spectrum disorders were characterized at single-cell resolution to gain insight into the pathophysiology of the disease. An atlas of the placenta at single cell resolution in accreta allows for understanding in the biology of the intimate maternal and fetal interaction. The contributions of stromal and endothelial cells were demonstrated through alterations in the extracellular matrix, growth factors, and angiogenesis. Transcriptional and protein changes in the stroma of placenta accreta spectrum shift the etiologic explanation away from \"invasive trophoblast\" to \"loss of boundary limits\" in the decidua. Gene targets identified in this study may be used to refine diagnostic assays in early pregnancy, track disease progression over time, and inform therapeutic discoveries.
目的:本研究旨在通过使用单细胞和空间分辨转录组学来表征胎盘植入谱系障碍中母胎界面的细胞异质性,从而深入了解对胎盘植入谱系障碍的理解。
方法:为了评估细胞异质性和细胞类型的功能,使用单细胞RNA测序和空间分辨转录组学。总共包括12个胎盘,6个胎盘伴胎盘植入谱系障碍和6个对照。对于每个胎盘植入谱系障碍,在以下部位进行了多次活检:同一胎盘中的胎盘植入谱贴壁和非贴壁部位。值得注意的是,2个平台用于生成库:用于单细胞和空间分辨转录组的10×Chromium和NanoStringGeoMX数字空间分析器,分别。使用一套生物信息学工具(Seurat和GeoMxToolsR包)进行差异基因表达分析。使用Clipper进行多次测试的校正。用RNAscope进行原位杂交,和免疫组织化学用于评估蛋白质表达。
结果:在创建胎盘植入细胞图谱时,在胎盘植入谱和对照组之间,活检部位的转录谱有显著差异。大多数差异是在遵守现场注意到的;然而,胎盘植入中同一胎盘的贴壁和非贴壁部位之间存在差异。在所有细胞类型中,内皮基质群体表现出最大的基因表达差异,由胶原蛋白基因的变化驱动,即III型胶原α1链(COL3A1),生长因子,表皮生长因子样蛋白6(EGFL6),和肝细胞生长因子(HGF),和血管生成相关基因,即δ样非规范Notch配体1(DLK1)和血小板内皮细胞粘附分子1(PECAM1)。胎盘内嗜性(同一胎盘中的粘附与非粘附位点)是由内皮基质细胞的差异驱动的,在胎盘植入谱的粘附与非粘附位点中,骨形态发生蛋白5(BMP5)和骨桥蛋白(SPP1)存在显着差异。
结论:以单细胞分辨率表征胎盘植入谱系障碍,以深入了解该疾病的病理生理学。植入中单细胞分辨率的胎盘图集可以理解母体和胎儿亲密相互作用的生物学。基质和内皮细胞的贡献通过细胞外基质的改变得到证实,生长因子,和血管生成。胎盘植入光谱基质的转录和蛋白质变化将病因解释从“侵入性滋养层”转移到蜕膜中的“边界界限丧失”。本研究中确定的基因靶标可用于改善妊娠早期的诊断测定,跟踪疾病随时间的进展,并告知治疗发现。