In humans and other mammalian species, damage to mechano-sensory hair cells is irreversible, leading to deafness and balance disorders. Remarkably, non-mammalian vertebrates regenerate lost hair cells. In birds, supporting cells have been shown to replace lost hair cells by mechanisms involving either cell division or direct trans-differentiation. In the mammalian auditory sensory organ, supporting cells do not regenerate lost hair cells. Strikingly, our findings suggest that mammalian supporting cells retain the intrinsic ability to function as hair cell progenitors, but their plasticity to regenerate hair cells is actively suppressed by external inhibitory cues. Our long-term goal is to utilize the latent plasticity of supporting cells to develop supporting cell based hair cell replacement strategies. To attain this goal, it is vital to improve our knowledge of the molecular programs active in developing and in mature supporting cells. We reason that to """"""""reprogram"""""""" and induce de- differentiation of supporting cells and consequently trans-differentiation of supporting cells into hair cells, we first have to understand the signals that control differentiation and maintenance in the supporting cell lineage. The goal of this proposal is to determine if the Notch signaling pathway, an evolutionary ancient and highly conserved cell-to-cell communication mechanism, functions in supporting cell differentiation and cell maintenance.
In Specific Aim 1 of our proposal, we will test if Notch signaling plays an instructive role in supporting cell differentiation. We will test if induction of an activated form of the Notch1 receptor is sufficient to induce supporting cell fate and whether inhibition of Notch signaling with 3-secretase inhibitors effects the onset and progression of supporting cell differentiation.
In Specific Aim 2 of our proposal, we will ablate Rbpj, a core component of the canonical Notch signaling pathway, in supporting cells to address if Notch signaling is required for supporting cell maintenance in the intact and hair cell damaged cochlea. To do so we will employ Rbpj conditional mouse mutants in combination with tamoxifen inducible CreERT lines. We anticipate that this analysis will provide valuable insight into the molecular mechanisms that drive supporting cell differentiation and elucidate the function of Notch signaling in supporting cell maintenance in the adult cochlea. In parallel, we will address a longstanding question-""""""""does persistence of Notch signaling in the hair cell damaged cochlea underlie the lack of hair cell regeneration in mammals?"""""""" Addressing this question is relevant to human health as inhibiting Notch signaling using 3-secretase inhibitors in mature cochlea could provide a means for restoring a latent capacity to regenerate hair cells.

Public Health Relevance

It is believed that lack of hair cell regeneration in mammals is due to an inability of neighboring supporting cells to regenerate damaged hair cells. In our proposal, we will characterize the molecular mechanisms instructing supporting cell differentiation and maintenance to elucidate why supporting cells fail to de-differentiate and regenerate lost hair cells. We believe that our findings will shed light on the molecular machinery that restrict hair cell regeneration and could provide new targets for future hair cell replacement therapies.

Agency
National Institute of Health (NIH)
Institute
National Institute on Deafness and Other Communication Disorders (NIDCD)
Type
Research Project (R01)
Project #
5R01DC011571-02
Application #
8233258
Study Section
Auditory System Study Section (AUD)
Program Officer
Freeman, Nancy
Project Start
2011-03-01
Project End
2016-02-29
Budget Start
2012-03-01
Budget End
2013-02-28
Support Year
2
Fiscal Year
2012
Total Cost
$410,000
Indirect Cost
$160,000
Name
Johns Hopkins University
Department
Neurosciences
Type
Schools of Medicine
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Doetzlhofer, Angelika; Avraham, Karen B (2017) Insights into inner ear-specific gene regulation: Epigenetics and non-coding RNAs in inner ear development and regeneration. Semin Cell Dev Biol 65:69-79
Basch, Martin L; Brown 2nd, Rogers M; Jen, Hsin-I et al. (2016) Fine-tuning of Notch signaling sets the boundary of the organ of Corti and establishes sensory cell fates. Elife 5:
Campbell, Dean P; Chrysostomou, Elena; Doetzlhofer, Angelika (2016) Canonical Notch signaling plays an instructive role in auditory supporting cell development. Sci Rep 6:19484
Golden, Erin J; Benito-Gonzalez, Ana; Doetzlhofer, Angelika (2015) The RNA-binding protein LIN28B regulates developmental timing in the mammalian cochlea. Proc Natl Acad Sci U S A 112:E3864-73
Korrapati, Soumya; Roux, Isabelle; Glowatzki, Elisabeth et al. (2013) Notch signaling limits supporting cell plasticity in the hair cell-damaged early postnatal murine cochlea. PLoS One 8:e73276