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

项目总结/摘要 听力损失是儿童发病的主要原因，在美国，每1，000人中约有2至3人患有听力损失。 美国.虽然对受影响基因的研究已经揭示了对机制的重要见解，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 本研究旨在阐明静纤毛的形成和维持的调控机制， 耳蜗毛细胞束，以及导致耳聋的这一过程中的缺陷。我们建议调查 Fam 65 b蛋白复合物在耳蜗毛细胞中的功能及听觉感受。在我们的初步调查中 通过研究，我们已经鉴定了Fam 65 b蛋白复合物中的几种组分。我们已经生成了基因组 使用CRISPR/Cas9系统修饰的小鼠模型。我们采用了免疫染色，并说明了 立纤毛缺陷。基于我们的初步数据，我们假设Fam 65 b蛋白 复合体对耳蜗毛细胞静纤毛束的结构组织至关重要。来测试我们 假设，我们将：i）功能性表征位于基底的Fam 65 b蛋白复合物中的蛋白质 ii）表征Fam 65 b蛋白复合物调节毛发的生化途径 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 Fam 65 b蛋白复合物中毛细胞蛋白的鉴定。我们的初步数据表明， 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， 形成静纤毛并决定其性质。很有可能，我们的发现可能会将几个与肥胖有关的 基因进入一个共同的分子途径，并提供新的线索，为发展的治疗 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.