The molecular identities of many essential components of the hair cell stereocilia in the inner ear are still unknown, precluding our understanding of the precise mechanisms of the mechanotransduction (MET) of sound. To this end, studies of the gene/protein determinants of Usher Syndrome (USH), a neurosensory disorder affecting both hearing and vision in humans, have been fruitful in elucidating the molecular components of transduction machinery in the inner ear. The long-term goal of our research is to understand fully how USH proteins are involved in the biological processes of the inner ear as a means of developing new strategies to prevent and treat this dual neurosensory disorder. The objective of this particular application is to determine the role of a newly identifid USH protein, CIB2, in stereocilia bundle formation, Ca2+ homeostasis and MET, as well as the mechanisms underlying hearing impairment caused by the loss of CIB2 function. The CIB2 protein is encoded by CIB2, the causative gene for Usher syndrome type 1 and non-syndromic deafness in 58 human families. CIB2 is localized at the upper part of stereocilia of both inner and outer hair cells in the mouse inner ear. Based on strong preliminary data, the central hypothesis of this proposal is that CIB2 is a Ca2+-buffering protein required for the maintenance of Ca2+ homeostasis in the mechanosensory stereocilia of the inner ear hair cells. Loss of CIB2 function, then, will affect both the structure and mechanosensitivity of stereocilia bundles, resulting in hearing loss. The rationale for the proposed research is that once the function of CIB2 in the inner ear is known, this knowledge will improve our understanding of auditory MET and Ca2+ homeostasis processes at the molecular level, which will ultimately aid in developing therapeutic agents for preventing or reversing hearing loss. Thus, the proposed research is relevant to that part of NIH's mission that pertains to developing fundamental knowledge that will potentially help to reduce the burdens of human disability. This hypothesis will be tested by pursuing three specific aims: (1) elucidate the mechanism of hearing loss in Cib2F91S mice, (2) elucidate the role of CIB2 in the development and function of stereocilia bundles, and (3) elucidate potential interactions between CIB2 and known USH proteins present in the inner ear hair cell stereocilia. Our experimental approach is to define the consequences of loss of CIB2 on inner ear morphology, the MET current, adaptation, Ca2+ concentration in the stereocilia, cytoskeletal and ultra-structural alterations. Our studies will employ contemporary genetic, molecular, cellular, histochemical and physiological techniques. The approach is innovative, because CIB2 is a novel USH protein, therefore, its role in the hearing and vision processes has never been investigated. The proposed research is significant, first, because it is expected to provide mechanistic understanding of how CIB2 is involved in the structural development and function of inner ear hair cell stereocilia. Secondly, the proposed study will likely uncover the mechanisms and physiological significance of Ca2+ buffering in stereocilia, which is critically important in auditory MET.

Public Health Relevance

Advancement in the understanding of molecular mechanisms of the inner ear is a prerequisite to developing therapeutic strategies for hearing impairment. The studies outlined in this proposal seek to understand the general function of a calcium binding protein known as CIB2 and its associated pathophysiology due to mutations of CIB2 encoding this protein. Thus the proposed research is developing fundamental knowledge that will help to reduce the burden of human hearing loss.

Agency
National Institute of Health (NIH)
Institute
National Institute on Deafness and Other Communication Disorders (NIDCD)
Type
Research Project (R01)
Project #
5R01DC012564-02
Application #
8573844
Study Section
Auditory System Study Section (AUD)
Program Officer
Watson, Bracie
Project Start
2012-12-01
Project End
2017-11-30
Budget Start
2013-12-01
Budget End
2014-11-30
Support Year
2
Fiscal Year
2014
Total Cost
$341,201
Indirect Cost
$83,345
Name
Cincinnati Children's Hospital Medical Center
Department
Type
DUNS #
071284913
City
Cincinnati
State
OH
Country
United States
Zip Code
45229
Giese, Arnaud P J; Tang, Yi-Quan; Sinha, Ghanshyam P et al. (2017) CIB2 interacts with TMC1 and TMC2 and is essential for mechanotransduction in auditory hair cells. Nat Commun 8:43
Dev Borman, Arundhati; Rachitskaya, Aleksandra; Suzani, Martina et al. (2017) Benign Yellow Dot Maculopathy: A New Macular Phenotype. Ophthalmology 124:1004-1013
Rehman, Atteeq U; Bird, Jonathan E; Faridi, Rabia et al. (2016) Mutational Spectrum of MYO15A and the Molecular Mechanisms of DFNB3 Human Deafness. Hum Mutat 37:991-1003
Fong, Keith S K; Hufnagel, Robert B; Khadka, Vedbar S et al. (2016) A mutation in the tuft mouse disrupts TET1 activity and alters the expression of genes that are crucial for neural tube closure. Dis Model Mech 9:585-96
Hufnagel, Robert B; Zimmerman, Sarah L; Krueger, Laura A et al. (2016) A new frontonasal dysplasia syndrome associated with deletion of the SIX2 gene. Am J Med Genet A 170A:487-91
Seco, Celia Zazo; Giese, Arnaud P; Shafique, Sobia et al. (2016) Novel and recurrent CIB2 variants, associated with nonsyndromic deafness, do not affect calcium buffering and localization in hair cells. Eur J Hum Genet 24:542-9
Waryah, A M; Shahzad, M; Shaikh, H et al. (2016) A novel CHST3 allele associated with spondyloepiphyseal dysplasia and hearing loss in Pakistani kindred. Clin Genet 90:90-5
Simon, Mariella; Richard, Elodie M; Wang, Xinjian et al. (2015) Mutations of human NARS2, encoding the mitochondrial asparaginyl-tRNA synthetase, cause nonsyndromic deafness and Leigh syndrome. PLoS Genet 11:e1005097
Rehman, Atteeq U; Santos-Cortez, Regie Lyn P; Drummond, Meghan C et al. (2015) Challenges and solutions for gene identification in the presence of familial locus heterogeneity. Eur J Hum Genet 23:1207-15
Mašindová, Ivica; Šoltýsová, Andrea; Varga, Lukáš et al. (2015) MARVELD2 (DFNB49) mutations in the hearing impaired Central European Roma population--prevalence, clinical impact and the common origin. PLoS One 10:e0124232

Showing the most recent 10 out of 22 publications