Department of Orthodontics, University of Texas Health Science Center School of Dentistry, Houston, TX, USA.
Department of Diagnostic and Biomedical Sciences, University of Texas Health Science Center School of Dentistry, Houston, TX, USA.
Orthod Craniofac Res. 2021 Nov;24(4):486-493. doi: 10.1111/ocr.12462. Epub 2021 Jan 7.
To determine the functional effects of ATF1, WNT10B and GREM2 gene variants identified in individuals with tooth agenesis (TA).
Stem cells from human exfoliated deciduous teeth (SHED) were used as an in vitro model system to test the effect of TA-associated variants.
Plasmid constructs containing reference and mutant alleles for ATF1 rs11169552, WNT10B rs833843 and GREM2 rs1414655 variants were transfected into SHED for functional characterization of variants. Allele-specific changes in gene transcription activity, protein expression, cell migration and proliferation, and expression of additional tooth development genes (MSX1, PAX9 and AXIN2) were evaluated. Data analyses were performed using Student's t-test. P-values ≤ .05 were considered statistically significant.
Mutant variants resulted in significantly decreased transcriptional activity of respective genes (P < 0.05), although no changes in protein localization were noted. Expression of MSX1 was significantly decreased in ATF1- and GREM2-mutant cells, whereas PAX9 or AXIN2 mRNA expression was not significantly altered. Mutant WNT10B had no significant effect on the expression of additional TA genes. ATF1- and GREM2-mutant cells presented increased cell migration. Cell proliferation was also affected with all three mutant alleles.
Our results demonstrate that ATF1, WNT10B and GREM2 mutant alleles have modulatory effects on gene/protein function that may contribute to TA.
确定在牙缺失（TA）个体中鉴定出的 ATF1、WNT10B 和 GREM2 基因变异的功能影响。
人脱落乳牙（SHED）干细胞被用作体外模型系统来测试与 TA 相关的变异的影响。
将包含 ATF1 rs11169552、WNT10B rs833843 和 GREM2 rs1414655 变异的参考和突变等位基因的质粒构建体转染到 SHED 中，以对变异进行功能表征。评估了基因转录活性、蛋白表达、细胞迁移和增殖以及额外的牙齿发育基因（MSX1、PAX9 和 AXIN2）的表达的等位基因特异性变化。使用学生 t 检验进行数据分析。P 值≤0.05 被认为具有统计学意义。
突变变异导致各自基因的转录活性显著降低（P<0.05），尽管未观察到蛋白定位的变化。ATF1 和 GREM2 突变细胞中 MSX1 的表达显著降低，而 PAX9 或 AXIN2 mRNA 表达没有显著改变。突变 WNT10B 对额外 TA 基因的表达没有显著影响。ATF1 和 GREM2 突变细胞表现出增加的细胞迁移。所有三种突变等位基因也影响细胞增殖。
我们的结果表明，ATF1、WNT10B 和 GREM2 突变等位基因对基因/蛋白功能具有调节作用，可能导致 TA。
