College of Health and Life Sciences, Hamad Bin Khalifa University (HBKU), Qatar Foundation (QF), Doha, Qatar.
Diabetes Research Center (DRC), Qatar Biomedical Research Institute (QBRI), Hamad Bin Khalifa University (HBKU), Qatar Foundation (QF), PO Box 34110, Doha, Qatar.
Stem Cell Rev Rep. 2023 May;19(4):1082-1097. doi: 10.1007/s12015-023-10515-3. Epub 2023 Feb 7.
Recently, we reported that forkhead box A2 (FOXA2) is required for the development of human pancreatic α- and β-cells. However, whether miRNAs play a role in regulating pancreatic genes during pancreatic development in the absence of FOXA2 expression is largely unknown. Here, we aimed to capture the dysregulated miRNAs and to identify their pancreatic-specific gene targets in pancreatic progenitors (PPs) derived from wild-type induced pluripotent stem cells (WT-iPSCs) and from iPSCs lacking FOXA2 (FOXA2iPSCs). To identify differentially expressed miRNAs (DEmiRs), and genes (DEGs), two different FOXA2iPSC lines were differentiated into PPs. FOXA2 PPs showed a significant reduction in the expression of the main PP transcription factors (TFs) in comparison to WT-PPs. RNA sequencing analysis demonstrated significant reduction in the mRNA expression of genes involved in the development and function of exocrine and endocrine pancreas. Furthermore, miRNA profiling identified 107 downregulated and 111 upregulated DEmiRs in FOXA2 PPs compared to WT-PPs. Target prediction analysis between DEmiRs and DEGs identified 92 upregulated miRNAs, predicted to target 1498 downregulated genes in FOXA2 PPs. Several important pancreatic TFs essential for pancreatic development were targeted by multiple DEmiRs. Selected DEmiRs and DEGs were further validated using RT-qPCR. Our findings revealed that FOXA2 expression is crucial for pancreatic development through regulating the expression of pancreatic endocrine and exocrine genes targeted by a set of miRNAs at the pancreatic progenitor stage. These data provide novel insights of the effect of FOXA2 deficiency on miRNA-mRNA regulatory networks controlling pancreatic development and differentiation.
最近，我们报道了叉头框 A2（FOXA2）在人类胰腺α和β细胞的发育中是必需的。然而，在缺乏 FOXA2 表达的情况下，miRNA 是否在调节胰腺发育过程中的胰腺基因方面发挥作用，在很大程度上尚不清楚。在这里，我们旨在捕获失调的 miRNA，并鉴定其在源自野生型诱导多能干细胞（WT-iPSCs）和缺乏 FOXA2 的 iPSCs（FOXA2iPSCs）的胰腺祖细胞（PPs）中的胰腺特异性基因靶标。为了鉴定差异表达的 miRNA（DEmiR）和基因（DEG），我们将两个不同的 FOXA2iPSC 系分化为 PPs。与 WT-PPs 相比，FOXA2 PPs 的主要 PP 转录因子（TFs）的表达显著降低。RNA 测序分析表明，参与外分泌和内分泌胰腺发育和功能的基因的 mRNA 表达显著降低。此外，miRNA 谱分析鉴定出 107 个下调和 111 个上调的 DEmiR 在 FOXA2 PPs 中与 WT-PPs 相比。DEmiR 和 DEG 之间的靶预测分析鉴定出 92 个上调的 miRNA，预测靶向 FOXA2 PPs 中 1498 个下调的基因。几个对胰腺发育至关重要的重要胰腺 TF 被多个 DEmiR 靶向。使用 RT-qPCR 进一步验证了选定的 DEmiR 和 DEG。我们的研究结果表明，FOXA2 表达通过调节一组在胰腺祖细胞阶段靶向胰腺内分泌和外分泌基因的 miRNA 的表达，对胰腺发育至关重要。这些数据提供了 FOXA2 缺乏对控制胰腺发育和分化的 miRNA-mRNA 调控网络的影响的新见解。
