Functions of Fam65b protein complex at the basal stereocilia in hearing and deafness

项目摘要/摘要 听力损失是儿童发病率的主要原因，在中国，每1000人中约有2至3人受到影响 美国。虽然对受影响的基因的研究已经为机制提供了重要的见解，但我们只是在 开始了解形态发生事件背后的疾病机制。的总目标是 本研究旨在阐明立体纤毛形成和维持的调控机制 耳蜗毛细胞束，以及在这一过程中导致耳聋的缺陷。我们建议调查 Fam65b蛋白复合体在耳蜗毛细胞和听觉中的作用。在我们的预赛中 研究表明，我们已经确定了Fam65b蛋白复合体中的几个成分。我们已经产生了基因组 用CRISPR/Cas9系统改良小鼠模型。我们已经使用了免疫染色并举例说明了 这些小鼠的立体纤毛缺陷。根据我们的初步数据，我们假设Fam65b蛋白 复合体对耳蜗毛细胞立体纤毛束的结构组织至关重要。测试我们的 假设，我们将：i)对位于碱基的Fam65b蛋白复合体中的蛋白质进行功能表征 立体纤毛区域；ii)表征Fam65b蛋白复合体调节头发的生化途径 细胞功能并确定它们在多大程度上调节立体纤毛中的肌动蛋白细胞骨架；iii)继续 我们鉴定了Fam65b蛋白复合体中的毛细胞蛋白。我们的初步数据表明了这种方法的可行性 我们的方法。我们预计，我们的研究将对分子机制提供新的见解 塑造立体纤毛并确定其属性。很有可能，我们的发现可能会将几个与耳聋有关的 基因转化为共同的分子途径，为治疗药物的发展提供了新的线索 治疗某些形式的疾病的方法。

PROJECT SUMMARY/ABSTRACT Hearing loss is a leading cause of morbidity in children affecting about 2 to 3 out of every 1,000 individuals in the USA. While the study of affected genes has shed important insights into the mechanisms, we are just at the beginning to understand the disease mechanisms underlying morphogenetic events. The overall goal of this research is to elucidate the mechanisms that regulate the formation and maintenance of the stereociliary bundle of cochlear hair cells, and the defects in this process that cause deafness. We propose to investigate the functions of Fam65b protein complex in the cochlear hair cells and auditory perceptions. In our preliminary studies, we have identified several components in the Fam65b protein complex. We have generated genomic modified mouse models using CRISPR/Cas9 system. We have employed immunostaining and illustrated the stereociliary defects in these mice. Based on our preliminary data, we hypothesize that Fam65b protein complex is critical for the structural organization of stereociliary bundle of cochlear hair cells. To test our hypothesis, we will: i) Functionally characterize proteins in the Fam65b protein complex localized at the basal region of stereocilia; ii) Characterize the biochemical pathways by which Fam65b protein complex regulate hair cell function and determine the extent to which they modulate the actin cytoskeleton in stereocilia; iii) Continue our identification of hair cell proteins in the Fam65b protein complex. Our preliminary data show the feasibility of our approach. We anticipate that our studies will shed new insights into the molecular machinery that shapes stereocilia and determines its properties. Quite possibly, our findings may link several deafness-related genes into a common molecular pathway and provide new leads for the development of therapeutic approaches for the treatment of some forms of the disease.