Loss of inner ear sensory cells is a major cause of permanent hearing loss. Our long-term goal is to provide a treatment option for patients suffering from hair cell loss by in vitro selection and characterization of progenitor cells that ultimately can be used for cell-replacement therapy of the damaged inner ear. We will select and characterize inner ear progenitor cells from two sources. First, we will optimize the isolation of progenitor cells derived from the adult mouse utricle (Aim 1). We have found in preliminary studies that the mouse utricle contains cells that display defining characteristics of progenitor cells, that these cells can be maintained in culture for several months, and that these cells can differentiate into cells that express immunological markers indicative of hair cells. A second source for progenitor cells is embryonic stem (ES) cells. In pilot studies, we have generated from mouse ES cells a population of cells that displayed expression of markers genes indicative of the developing inner ear.
Our second Aim i s to establish in vitro conditions that enable us to routinely guide ES cell derivatives towards a progenitor cell phenotype that can be differentiated into mature inner ear cell types, which can be functionally characterized by in vitro and in vivo studies.
In Aim 3, we propose testing whether expression of the transcription factor Math1 in stem cells, including the progenitor cells generated with the experiments proposed in this application, can lead to increase in the number of generated hair cells. Finally, we will perform basic analyses of Features of progenitor cell-derived hair cell-like cells. We will test whether in vitro-generated hair cell-like cells do selectively label with the styryl dye FM1-43 and whether they display electrophysiological features of native hair cells (Aim 4A).
In Aim 4 B, we will generate major inner ear cell types in vivo by grafting presumptive progenitor cells into the developing chicken inner ear. ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute on Deafness and Other Communication Disorders (NIDCD)
Type
Research Project (R01)
Project #
7R01DC006167-04
Application #
7153765
Study Section
Special Emphasis Panel (ZDC1-SRB-J (11))
Program Officer
Freeman, Nancy
Project Start
2003-05-15
Project End
2008-04-30
Budget Start
2005-10-01
Budget End
2006-04-30
Support Year
4
Fiscal Year
2005
Total Cost
$223,832
Indirect Cost
Name
Stanford University
Department
Otolaryngology
Type
Schools of Medicine
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94305
Lee, Jiyoon; B?scke, Robert; Tang, Pei-Ciao et al. (2018) Hair Follicle Development in Mouse Pluripotent Stem Cell-Derived Skin Organoids. Cell Rep 22:242-254
Durruthy-Durruthy, Robert; Gottlieb, Assaf; Heller, Stefan (2015) 3D computational reconstruction of tissues with hollow spherical morphologies using single-cell gene expression data. Nat Protoc 10:459-474
Waldhaus, Jörg; Durruthy-Durruthy, Robert; Heller, Stefan (2015) Quantitative High-Resolution Cellular Map of the Organ of Corti. Cell Rep 11:1385-99
Durruthy-Durruthy, Robert; Heller, Stefan (2015) Applications for single cell trajectory analysis in inner ear development and regeneration. Cell Tissue Res 361:49-57
Hartman, Byron H; Durruthy-Durruthy, Robert; Laske, Roman D et al. (2015) Identification and characterization of mouse otic sensory lineage genes. Front Cell Neurosci 9:79
Durruthy-Durruthy, Robert; Gottlieb, Assaf; Hartman, Byron H et al. (2014) Reconstruction of the mouse otocyst and early neuroblast lineage at single-cell resolution. Cell 157:964-78
Ronaghi, Mohammad; Nasr, Marjan; Ealy, Megan et al. (2014) Inner ear hair cell-like cells from human embryonic stem cells. Stem Cells Dev 23:1275-84
Volkenstein, Stefan; Oshima, Kazuo; Sinkkonen, Saku T et al. (2013) Transient, afferent input-dependent, postnatal niche for neural progenitor cells in the cochlear nucleus. Proc Natl Acad Sci U S A 110:14456-61
Jan, Taha Adnan; Chai, Renjie; Sayyid, Zahra Nabi et al. (2013) Tympanic border cells are Wnt-responsive and can act as progenitors for postnatal mouse cochlear cells. Development 140:1196-206
Ronaghi, Mohammad; Nasr, Marjan; Heller, Stefan (2012) Concise review: Inner ear stem cells--an oxymoron, but why? Stem Cells 30:69-74

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