Mellott Adam J, Devarajan Keerthana, Shinogle Heather E, Moore David S, Talata Zsolt, Laurence Jennifer S, Forrest M Laird, Noji Sumihare, Tanaka Eiji, Staecker Hinrich, Detamore Michael S
1Bioengineering Graduate Program, University of Kansas, Lawrence, Kansas.
2Department of Genetics, Stanford University, Stanford, California.
Tissue Eng Part A. 2015 Jun;21(11-12):1795-809. doi: 10.1089/ten.TEA.2014.0340. Epub 2015 Apr 13.
The transcription factor atonal homolog 1 (ATOH1) has multiple homologues that are functionally conserved across species and is responsible for the generation of sensory hair cells. To evaluate potential functional differences between homologues, human and mouse ATOH1 (HATH1 and MATH-1, respectively) were nonvirally delivered to human Wharton's jelly cells (hWJCs) for the first time. Delivery of HATH1 to hWJCs demonstrated superior expression of inner ear hair cell markers and characteristics than delivery of MATH-1. Inhibition of HES1 and HES5 signaling further increased the atonal effect. Transfection of hWJCs with HATH1 DNA, HES1 siRNA, and HES5 siRNA displayed positive identification of key hair cell and support cell markers found in the cochlea, as well as a variety of cell shapes, sizes, and features not native to hair cells, suggesting the need for further examination of other cell types induced by HATH1 expression. In the first side-by-side evaluation of HATH1 and MATH-1 in human cells, substantial differences were observed, suggesting that the two atonal homologues may not be interchangeable in human cells, and artificial expression of HATH1 in hWJCs requires further study. In the future, this line of research may lead to engineered systems that would allow for evaluation of drug ototoxicity or potentially even direct therapeutic use.
转录因子无调性同源物1(ATOH1)有多个同源物,它们在物种间功能保守,负责感觉毛细胞的生成。为评估同源物之间的潜在功能差异,首次将人源和小鼠源的ATOH1(分别为HATH1和MATH-1)非病毒递送至人脐带华通氏胶细胞(hWJC)。将HATH1递送至hWJC显示出比递送MATH-1时内耳毛细胞标志物的表达及特征更优。抑制HES1和HES5信号通路进一步增强了无调性效应。用HATH1 DNA、HES1小干扰RNA(siRNA)和HES5 siRNA转染hWJC显示出耳蜗中关键毛细胞和支持细胞标志物的阳性鉴定,以及多种非毛细胞天然具有的细胞形状、大小和特征,这表明需要进一步研究由HATH1表达诱导的其他细胞类型。在首次对人细胞中的HATH1和MATH-1进行的并列评估中,观察到显著差异,表明这两种无调性同源物在人细胞中可能不可互换,且在hWJC中人工表达HATH1需要进一步研究。未来,这一系列研究可能会带来可用于评估药物耳毒性甚至可能直接用于治疗的工程系统。