Department of Diagnostic & Biomedical Sciences, School of Dentistry, The University of Texas Health Science Center at Houston, Houston, TX 77054, USA.
Center for Craniofacial Research, The University of Texas Health Science Center at Houston, Houston, TX 77054, USA.
Int J Mol Sci. 2021 Nov 18;22(22):12453. doi: 10.3390/ijms222212453.
Cleft lip with or without cleft palate (CL/P) is one of the most common congenital birth defects. This study aims to identify novel pathogenic microRNAs associated with cleft palate (CP). Through data analyses of miRNA-sequencing for developing palatal shelves of C57BL/6J mice, we found that miR-449a-3p, miR-449a-5p, miR-449b, miR-449c-3p, and miR-449c-5p were significantly upregulated, and that miR-19a-3p, miR-130a-3p, miR-301a-3p, and miR-486b-5p were significantly downregulated, at embryonic day E14.5 compared to E13.5. Among them, overexpression of the miR-449 family (miR-449a-3p, miR-449a-5p, miR-449b, miR-449c-3p, and miR-449c-5p) and miR-486b-5p resulted in reduced cell proliferation in primary mouse embryonic palatal mesenchymal (MEPM) cells and mouse cranial neural crest cell line O9-1. On the other hand, inhibitors of miR-130a-3p and miR-301a-3p significantly reduced cell proliferation in MEPM and O9-1 cells. Notably, we found that treatment with dexamethasone, a glucocorticoid known to induce CP in mice, suppressed miR-130a-3p expression in both MEPM and O9-1 cells. Moreover, a miR-130a-3p mimic could ameliorate the cell proliferation defect induced by dexamethasone through normalization of expression. Taken together, our results suggest that miR-130-3p plays a crucial role in dexamethasone-induced CP in mice.
唇裂伴或不伴腭裂（CL/P）是最常见的先天性出生缺陷之一。本研究旨在鉴定与腭裂（CP）相关的新型致病微小 RNA（miRNA）。通过对 C57BL/6J 小鼠发育中腭突的 miRNA 测序数据分析，我们发现 miR-449a-3p、miR-449a-5p、miR-449b、miR-449c-3p 和 miR-449c-5p 在胚胎第 14.5 天（E14.5）时比第 13.5 天（E13.5）显著上调，而 miR-19a-3p、miR-130a-3p、miR-301a-3p 和 miR-486b-5p 则显著下调。其中，miR-449 家族（miR-449a-3p、miR-449a-5p、miR-449b、miR-449c-3p 和 miR-449c-5p）和 miR-486b-5p 的过表达导致原代小鼠胚胎腭中胚层（MEPM）细胞和小鼠颅神经嵴细胞系 O9-1 的细胞增殖减少。另一方面，miR-130a-3p 和 miR-301a-3p 的抑制剂显著减少了 MEPM 和 O9-1 细胞的增殖。值得注意的是，我们发现，已知可诱导小鼠 CP 的糖皮质激素地塞米松处理可抑制 MEPM 和 O9-1 细胞中 miR-130a-3p 的表达。此外，miR-130a-3p 模拟物可通过恢复 表达来改善地塞米松诱导的细胞增殖缺陷。综上所述，我们的研究结果表明，miR-130a-3p 在糖皮质激素诱导的小鼠 CP 中发挥关键作用。
