Department of Orofacial Sciences, School of Dentistry, University of California, San Francisco, 521 Parnasus Ave, Box 0422, San Francisco, CA, 04143-0422, USA.
Sci Rep. 2022 Feb 18;12(1):2820. doi: 10.1038/s41598-022-06751-1.
As the hardest tissue in the human body, tooth enamel formation is a highly regulated process involving several stages of differentiation and key regulatory genes. One such gene, tryptophan-aspartate repeat domain 72 (WDR72), has been found to cause a tooth enamel defect when deleted or mutated, resulting in a condition called amelogenesis imperfecta. Unlike the canonical genes regulating tooth development, WDR72 remains intracellularly and is not secreted to the enamel matrix space to regulate mineralization, and is found in other major organs of the body, namely the kidney, brain, liver, and heart. To date, a link between intracellular vesicle transport and enamel mineralization has been suggested, however identification of the mechanistic regulators has yet to be elucidated, in part due to the limitations associated with studying highly differentiated ameloblast cells. Here we show compelling evidence that WDR72 regulates endocytosis of proteins, both in vivo and in a novel in vitro ameloblast cell line. We elucidate WDR72's function to be independent of intracellular vesicle acidification while still leading to defective enamel matrix pH extracellularly. We identify a vesicle function associated with microtubule assembly and propose that WDR72 directs microtubule assembly necessary for membrane mobilization and subsequent vesicle transport. Understanding WDR72 function provides a mechanistic basis for determining physiologic and pathologic tissue mineralization.
作为人体中最坚硬的组织,牙釉质的形成是一个高度调控的过程,涉及几个分化阶段和关键的调节基因。其中一个基因,色氨酸-天冬氨酸重复结构域 72(WDR72),当其缺失或突变时会导致牙釉质缺陷,从而导致釉质不全症。与调节牙齿发育的典型基因不同,WDR72 保持在细胞内,不会分泌到釉基质空间来调节矿化,并且在身体的其他主要器官中也有发现,如肾脏、大脑、肝脏和心脏。迄今为止,已经提出了细胞内囊泡运输与牙釉质矿化之间的联系,然而,机制调节因子的鉴定尚未阐明,部分原因是由于研究高度分化的成釉细胞存在局限性。在这里,我们提供了令人信服的证据表明,WDR72 调节蛋白质的内吞作用,无论是在体内还是在新型体外成釉细胞瘤系中。我们阐明了 WDR72 的功能不依赖于细胞内囊泡酸化,但仍然导致细胞外釉基质 pH 缺陷。我们确定了与微管组装相关的囊泡功能,并提出 WDR72 指导微管组装,这对于膜动员和随后的囊泡运输是必要的。了解 WDR72 的功能为确定生理和病理组织矿化提供了机制基础。