Characterizing Adult Cochlear Supporting Cell Transcriptional Diversity Using Single-Cell RNA-Seq: Validation in the Adult Mouse and Translational Implications for the Adult Human Cochlea
Hearing loss is a problem that impacts a significant proportion of the adult population. Cochlear hair cell (HC) loss due to loud noise, chemotherapy and aging is the major underlying cause. A significant proportion of these individuals are dissatisfied with available treatment options which include...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2020-02-01
|
| Series: | Frontiers in Molecular Neuroscience |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fnmol.2020.00013/full |
| _version_ | 1819052463500558336 |
|---|---|
| author | Michael Hoa Rafal Olszewski Xiaoyi Li Ian Taukulis Shoujun Gu Alvin DeTorres Ivan A. Lopez Ivan A. Lopez Fred H. Linthicum Jr. Fred H. Linthicum Jr. Akira Ishiyama Akira Ishiyama Daniel Martin Robert J. Morell Matthew W. Kelley |
| author_facet | Michael Hoa Rafal Olszewski Xiaoyi Li Ian Taukulis Shoujun Gu Alvin DeTorres Ivan A. Lopez Ivan A. Lopez Fred H. Linthicum Jr. Fred H. Linthicum Jr. Akira Ishiyama Akira Ishiyama Daniel Martin Robert J. Morell Matthew W. Kelley |
| author_sort | Michael Hoa |
| collection | DOAJ |
| description | Hearing loss is a problem that impacts a significant proportion of the adult population. Cochlear hair cell (HC) loss due to loud noise, chemotherapy and aging is the major underlying cause. A significant proportion of these individuals are dissatisfied with available treatment options which include hearing aids and cochlear implants. An alternative approach to restore hearing would be to regenerate HCs. Such therapy would require a recapitulation of the complex architecture of the organ of Corti, necessitating regeneration of both mature HCs and supporting cells (SCs). Transcriptional profiles of the mature cell types in the cochlea are necessary to can provide a metric for eventual regeneration therapies. To assist in this effort, we sought to provide the first single-cell characterization of the adult cochlear SC transcriptome. We performed single-cell RNA-Seq on FACS-purified adult cochlear SCs from the LfngEGFP adult mouse, in which SCs express GFP. We demonstrate that adult cochlear SCs are transcriptionally distinct from their perinatal counterparts. We establish cell-type-specific adult cochlear SC transcriptome profiles, and we validate these expression profiles through a combination of both fluorescent immunohistochemistry and in situ hybridization co-localization and quantitative polymerase chain reaction (qPCR) of adult cochlear SCs. Furthermore, we demonstrate the relevance of these profiles to the adult human cochlea through immunofluorescent human temporal bone histopathology. Finally, we demonstrate cell cycle regulator expression in adult SCs and perform pathway analyses to identify potential mechanisms for facilitating mitotic regeneration (cell proliferation, differentiation, and eventually regeneration) in the adult mammalian cochlea. Our findings demonstrate the importance of characterizing mature as opposed to perinatal SCs. |
| first_indexed | 2024-12-21T12:20:14Z |
| format | Article |
| id | doaj.art-5259881393c44f4b90aaac30de669df3 |
| institution | Directory Open Access Journal |
| issn | 1662-5099 |
| language | English |
| last_indexed | 2024-12-21T12:20:14Z |
| publishDate | 2020-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Neuroscience |
| spelling | doaj.art-5259881393c44f4b90aaac30de669df32022-12-21T19:04:19ZengFrontiers Media S.A.Frontiers in Molecular Neuroscience1662-50992020-02-011310.3389/fnmol.2020.00013491389Characterizing Adult Cochlear Supporting Cell Transcriptional Diversity Using Single-Cell RNA-Seq: Validation in the Adult Mouse and Translational Implications for the Adult Human CochleaMichael Hoa0Rafal Olszewski1Xiaoyi Li2Ian Taukulis3Shoujun Gu4Alvin DeTorres5Ivan A. Lopez6Ivan A. Lopez7Fred H. Linthicum Jr.8Fred H. Linthicum Jr.9Akira Ishiyama10Akira Ishiyama11Daniel Martin12Robert J. Morell13Matthew W. Kelley14Auditory Restoration and Development Program, National Institute on Deafness and Other Communication Disorders, NIH, Bethesda, MD, United StatesAuditory Restoration and Development Program, National Institute on Deafness and Other Communication Disorders, NIH, Bethesda, MD, United StatesAuditory Restoration and Development Program, National Institute on Deafness and Other Communication Disorders, NIH, Bethesda, MD, United StatesAuditory Restoration and Development Program, National Institute on Deafness and Other Communication Disorders, NIH, Bethesda, MD, United StatesAuditory Restoration and Development Program, National Institute on Deafness and Other Communication Disorders, NIH, Bethesda, MD, United StatesAuditory Restoration and Development Program, National Institute on Deafness and Other Communication Disorders, NIH, Bethesda, MD, United StatesNational Temporal Bone Laboratory at UCLA, UCLA School of Medicine, Los Angeles, CA, United StatesCellular and Molecular Biology of the Inner Ear Laboratory, UCLA School of Medicine, Los Angeles, CA, United StatesNational Temporal Bone Laboratory at UCLA, UCLA School of Medicine, Los Angeles, CA, United StatesCellular and Molecular Biology of the Inner Ear Laboratory, UCLA School of Medicine, Los Angeles, CA, United StatesNational Temporal Bone Laboratory at UCLA, UCLA School of Medicine, Los Angeles, CA, United StatesCellular and Molecular Biology of the Inner Ear Laboratory, UCLA School of Medicine, Los Angeles, CA, United StatesBiomedical Research Informatics Office, National Institute of Dental and Craniofacial Research, NIH, Bethesda, MD, United StatesGenomics and Computational Biology Core, National Institute on Deafness and Other Communication Disorders, NIH, Bethesda, MD, United StatesLaboratory of Cochlear Development, National Institute on Deafness and Other Communication Disorders, NIH, Bethesda, MD, United StatesHearing loss is a problem that impacts a significant proportion of the adult population. Cochlear hair cell (HC) loss due to loud noise, chemotherapy and aging is the major underlying cause. A significant proportion of these individuals are dissatisfied with available treatment options which include hearing aids and cochlear implants. An alternative approach to restore hearing would be to regenerate HCs. Such therapy would require a recapitulation of the complex architecture of the organ of Corti, necessitating regeneration of both mature HCs and supporting cells (SCs). Transcriptional profiles of the mature cell types in the cochlea are necessary to can provide a metric for eventual regeneration therapies. To assist in this effort, we sought to provide the first single-cell characterization of the adult cochlear SC transcriptome. We performed single-cell RNA-Seq on FACS-purified adult cochlear SCs from the LfngEGFP adult mouse, in which SCs express GFP. We demonstrate that adult cochlear SCs are transcriptionally distinct from their perinatal counterparts. We establish cell-type-specific adult cochlear SC transcriptome profiles, and we validate these expression profiles through a combination of both fluorescent immunohistochemistry and in situ hybridization co-localization and quantitative polymerase chain reaction (qPCR) of adult cochlear SCs. Furthermore, we demonstrate the relevance of these profiles to the adult human cochlea through immunofluorescent human temporal bone histopathology. Finally, we demonstrate cell cycle regulator expression in adult SCs and perform pathway analyses to identify potential mechanisms for facilitating mitotic regeneration (cell proliferation, differentiation, and eventually regeneration) in the adult mammalian cochlea. Our findings demonstrate the importance of characterizing mature as opposed to perinatal SCs.https://www.frontiersin.org/article/10.3389/fnmol.2020.00013/fullinner earsupporting cell subtypessmFISHadult (MeSH)cell cycleFACS |
| spellingShingle | Michael Hoa Rafal Olszewski Xiaoyi Li Ian Taukulis Shoujun Gu Alvin DeTorres Ivan A. Lopez Ivan A. Lopez Fred H. Linthicum Jr. Fred H. Linthicum Jr. Akira Ishiyama Akira Ishiyama Daniel Martin Robert J. Morell Matthew W. Kelley Characterizing Adult Cochlear Supporting Cell Transcriptional Diversity Using Single-Cell RNA-Seq: Validation in the Adult Mouse and Translational Implications for the Adult Human Cochlea Frontiers in Molecular Neuroscience inner ear supporting cell subtypes smFISH adult (MeSH) cell cycle FACS |
| title | Characterizing Adult Cochlear Supporting Cell Transcriptional Diversity Using Single-Cell RNA-Seq: Validation in the Adult Mouse and Translational Implications for the Adult Human Cochlea |
| title_full | Characterizing Adult Cochlear Supporting Cell Transcriptional Diversity Using Single-Cell RNA-Seq: Validation in the Adult Mouse and Translational Implications for the Adult Human Cochlea |
| title_fullStr | Characterizing Adult Cochlear Supporting Cell Transcriptional Diversity Using Single-Cell RNA-Seq: Validation in the Adult Mouse and Translational Implications for the Adult Human Cochlea |
| title_full_unstemmed | Characterizing Adult Cochlear Supporting Cell Transcriptional Diversity Using Single-Cell RNA-Seq: Validation in the Adult Mouse and Translational Implications for the Adult Human Cochlea |
| title_short | Characterizing Adult Cochlear Supporting Cell Transcriptional Diversity Using Single-Cell RNA-Seq: Validation in the Adult Mouse and Translational Implications for the Adult Human Cochlea |
| title_sort | characterizing adult cochlear supporting cell transcriptional diversity using single cell rna seq validation in the adult mouse and translational implications for the adult human cochlea |
| topic | inner ear supporting cell subtypes smFISH adult (MeSH) cell cycle FACS |
| url | https://www.frontiersin.org/article/10.3389/fnmol.2020.00013/full |
| work_keys_str_mv | AT michaelhoa characterizingadultcochlearsupportingcelltranscriptionaldiversityusingsinglecellrnaseqvalidationintheadultmouseandtranslationalimplicationsfortheadulthumancochlea AT rafalolszewski characterizingadultcochlearsupportingcelltranscriptionaldiversityusingsinglecellrnaseqvalidationintheadultmouseandtranslationalimplicationsfortheadulthumancochlea AT xiaoyili characterizingadultcochlearsupportingcelltranscriptionaldiversityusingsinglecellrnaseqvalidationintheadultmouseandtranslationalimplicationsfortheadulthumancochlea AT iantaukulis characterizingadultcochlearsupportingcelltranscriptionaldiversityusingsinglecellrnaseqvalidationintheadultmouseandtranslationalimplicationsfortheadulthumancochlea AT shoujungu characterizingadultcochlearsupportingcelltranscriptionaldiversityusingsinglecellrnaseqvalidationintheadultmouseandtranslationalimplicationsfortheadulthumancochlea AT alvindetorres characterizingadultcochlearsupportingcelltranscriptionaldiversityusingsinglecellrnaseqvalidationintheadultmouseandtranslationalimplicationsfortheadulthumancochlea AT ivanalopez characterizingadultcochlearsupportingcelltranscriptionaldiversityusingsinglecellrnaseqvalidationintheadultmouseandtranslationalimplicationsfortheadulthumancochlea AT ivanalopez characterizingadultcochlearsupportingcelltranscriptionaldiversityusingsinglecellrnaseqvalidationintheadultmouseandtranslationalimplicationsfortheadulthumancochlea AT fredhlinthicumjr characterizingadultcochlearsupportingcelltranscriptionaldiversityusingsinglecellrnaseqvalidationintheadultmouseandtranslationalimplicationsfortheadulthumancochlea AT fredhlinthicumjr characterizingadultcochlearsupportingcelltranscriptionaldiversityusingsinglecellrnaseqvalidationintheadultmouseandtranslationalimplicationsfortheadulthumancochlea AT akiraishiyama characterizingadultcochlearsupportingcelltranscriptionaldiversityusingsinglecellrnaseqvalidationintheadultmouseandtranslationalimplicationsfortheadulthumancochlea AT akiraishiyama characterizingadultcochlearsupportingcelltranscriptionaldiversityusingsinglecellrnaseqvalidationintheadultmouseandtranslationalimplicationsfortheadulthumancochlea AT danielmartin characterizingadultcochlearsupportingcelltranscriptionaldiversityusingsinglecellrnaseqvalidationintheadultmouseandtranslationalimplicationsfortheadulthumancochlea AT robertjmorell characterizingadultcochlearsupportingcelltranscriptionaldiversityusingsinglecellrnaseqvalidationintheadultmouseandtranslationalimplicationsfortheadulthumancochlea AT matthewwkelley characterizingadultcochlearsupportingcelltranscriptionaldiversityusingsinglecellrnaseqvalidationintheadultmouseandtranslationalimplicationsfortheadulthumancochlea |