HAND1 knockdown disrupts trophoblast global gene expression.
HAND1 knockdown disrupts trophoblast global gene expression.
复制标题
DOI：
10.14814/phy2.15553
复制
发表时间：
2023-01
影响因子：
2.5
通讯作者：
中科院分区：
文献类型：
--
作者：
先天性心脏病(CHD)每年影响近1%的新生儿，先天性心脏病妊娠增加了发生胎盘异常病理的风险。我们之前曾报道，在多种小鼠模型中，干扰与CHD相关的基因Hand1在胎盘滋养层细胞和内皮细胞中的表达对发育有显著影响。在这项研究中，我们的目的是建立在这一知识的基础上，并表征干扰HAND1对人胎盘滋养层细胞和血管内皮细胞基因表达的机制影响。用HAND1siRNA沉默人绒毛膜滋养层细胞模型BeWo细胞(n=3~9代)和人胎盘微血管内皮细胞(HPMVEC；n=n=3代)96 h，收集细胞，分离基因，用Illumina NextSeq550测序平台进行测序。使用Edger软件包中的一般线性模型进行归一化和差异基因表达分析。使用>；1.0或 <； −1.0的log2倍变化和未调整的p值≤0.05来确定统计学意义。用Panther DB进行过表达分析，用字符串DB进行蛋白质关联网络分析。在直接敲除HAND1的BeWo细胞中，有664个基因下调，59个基因上调。过度表达分析确定了包括细胞分化、定位和细胞投射组织在内的通路的中断。相比之下，在HPMVEC中，只有7个基因在HAND1直接敲除的情况下发生了变化。阻断HAND1的表达会显著改变滋养层细胞的基因表达谱，但不会改变内皮细胞的表达谱。这一数据提供了进一步的证据，表明未来对先天性心脏病遗传扰动的研究除了考虑胎儿心脏外，还应该考虑胚胎外组织。胎盘滋养层细胞BeWo细胞模型中HAND1基因敲除基因的网络分析。灰色线条表示网络边缘，其粗细表示数据支持的置信度(较粗的线条=较高的置信度数据)。颜色编码的图例显示了可能受HAND1基因敲除影响的反应组途径中的基因。
Congenital heart disease (CHD) affects nearly 1% of births annually, and CHD pregnancies carry increased risk of developing pathologies of abnormal placentation. We previously reported significant developmental impacts of disrupting Hand1, a gene associated with CHD, expression in placenta trophoblast and endothelial cells in multiple mouse models. In this study, we aimed to build upon this knowledge and characterize the mechanistic impacts of disrupting HAND1 on human placenta trophoblast and vascular endothelial cell gene expression. HAND1 gene expression was silenced in BeWo cells, a choriocarcinoma model of human cytotrophoblasts, (n = 3–9 passages) and isolated human placental microvascular endothelial cells (HPMVEC; n = 3 passages), with HAND1 siRNA for 96 h. Cells were harvested, mRNA isolated and RNA sequencing performed using the Illumina NextSeq 550 platform. Normalization and differential gene expression analyses were conducted using general linear modeling in edgeR packages. Statistical significance was determined using a log2 fold change of >1.0 or < −1.0 and unadjusted p‐value ≤0.05. Panther DB was used for overrepresentation analysis, and String DB for protein association network analysis. There was downregulation of 664 genes, and upregulation of 59 genes in BeWo cells with direct HAND1 knockdown. Overrepresentation analysis identified disruption to pathways including cell differentiation, localization, and cell projection organization. In contrast, only seven genes were changed with direct HAND1 knockdown in HPMVECs. Disruption to HAND1 expression significantly alters gene expression profile in trophoblast but not endothelial cells. This data provides further evidence that future studies on genetic perturbations in CHDs should consider the extra‐embryonic tissue in addition to the fetal heart. Network analyses of genes disrupted by HAND1 knockdown in the placenta trophoblast BeWo cell line model. Gray lines represent the network edges with thickness representing the confidence of the data support (thicker lines = higher confidence data). Color coded legends show genes in Reactome pathways that may be impacted by HAND1 knockdown.
影响因子：
2.7
作者：
Tararbit K;Lelong N;Goffinet F;Khoshnood B;EPICARD Study Group
通讯作者：
EPICARD Study Group