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

项目概要/摘要 听力损失是儿童发病的主要原因，每 1,000 人中约有 2 至 3 人受到影响。 美国。虽然对受影响基因的研究已经为机制提供了重要的见解，但我们还处于起步阶段。 开始了解形态发生事件背后的疾病机制。总体目标为 这项研究旨在阐明调节静纤毛形成和维持的机制 耳蜗毛细胞束，以及该过程中的缺陷导致耳聋。我们建议调查 Fam65b 蛋白复合物在耳蜗毛细胞和听觉感知中的功能。在我们的初步 通过研究，我们鉴定了 Fam65b 蛋白复合物中的几种成分。我们已经生成了基因组 使用 CRISPR/Cas9 系统修改小鼠模型。我们采用了免疫染色并说明了 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 复合体对于耳蜗毛细胞静纤毛束的结构组织至关重要。来测试我们的 假设，我们将： i) 对位于基底的 Fam65b 蛋白复合物中的蛋白进行功能表征 静纤毛区域； ii) 表征 Fam65b 蛋白复合物调节头发的生化途径 细胞功能并确定它们调节静纤毛中肌动蛋白细胞骨架的程度； iii) 继续 我们对 Fam65b 蛋白复合物中毛细胞蛋白的鉴定。我们的初步数据显示了可行性 我们的方法。我们预计我们的研究将为分子机制提供新的见解 塑造静纤毛并决定其特性。我们的发现很可能与几种与耳聋相关的疾病有关 基因转化为共同的分子途径，为治疗药物的开发提供新的线索 治疗某些疾病的方法。

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.