Generating high-fidelity cochlear organoids from human pluripotent stem cells

Stephen T. Moore; Takashi Nakamura; Jing Nie; Alexander J. Solivais; Isabel Aristizábal-Ramírez; Yoshitomo Ueda; Mayakannan Manikandan; V. Shweta Reddy; Daniel R. Romano; John R. Hoffman; Benjamin J. Perrin; Rick F. Nelson; Gregory I. Frolenkov; Susana M. Chuva de Sousa Lopes; Eri Hashino

doi:10.1016/j.stem.2023.06.006

Generating high-fidelity cochlear organoids from human pluripotent stem cells

Stephen T. Moore
, Takashi Nakamura
, Jing Nie
, Alexander J. Solivais
, Isabel Aristizábal-Ramírez
, Yoshitomo Ueda
, Mayakannan Manikandan
, V. Shweta Reddy
, Daniel R. Romano
, John R. Hoffman
, Benjamin J. Perrin
, Rick F. Nelson
, Gregory I. Frolenkov
, Susana M. Chuva de Sousa Lopes
, Eri Hashino

Research output: Contribution to journal › Article › peer-review

53 Scopus citations

Abstract

Mechanosensitive hair cells in the cochlea are responsible for hearing but are vulnerable to damage by genetic mutations and environmental insults. The paucity of human cochlear tissues makes it difficult to study cochlear hair cells. Organoids offer a compelling platform to study scarce tissues in vitro; however, derivation of cochlear cell types has proven non-trivial. Here, using 3D cultures of human pluripotent stem cells, we sought to replicate key differentiation cues of cochlear specification. We found that timed modulations of Sonic Hedgehog and WNT signaling promote ventral gene expression in otic progenitors. Ventralized otic progenitors subsequently give rise to elaborately patterned epithelia containing hair cells with morphology, marker expression, and functional properties consistent with both outer and inner hair cells in the cochlea. These results suggest that early morphogenic cues are sufficient to drive cochlear induction and establish an unprecedented system to model the human auditory organ.

Original language	English
Pages (from-to)	950-961.e7
Journal	Cell Stem Cell
Volume	30
Issue number	7
DOIs	https://doi.org/10.1016/j.stem.2023.06.006
State	Published - Jul 6 2023

Bibliographical note

Publisher Copyright:
© 2023 Elsevier Inc.

Funding

This work was supported by a Department of Defense US Army Medical Research and Materiel Command Congressionally Directed Medical Research Program grant W81XWH-18-1-0062 (E. H.), National Institutes of Health grants R01 DC013294 and R01 DC015788 (E.H.), K08 DC016034 (R.F.N.), JSPS KAKENHI grant JP21K09638 (T.N.), the Indiana University Health-Indiana University School of Medicine Strategic Research Initiative (E.H.), and the Ruth C. Holton Research Funds (E.H.). The authors would like to thank the Center for Contraception, Sexuality, and Abortion (CASA) in Leiden and The Hague for the collection of the human fetal material; B. Daye, T. van der Helm, F. Wei, J. Harkin, T. Saeki, M. Jafarkkhani, E. Longworth-Mills, P. McGuire, and H. Gao for their expert technical assistance; J. Zheng and K. Homma for antibodies against PRESTIN; and M. W. Kelley for his comments on the manuscript. Conceptualization, E.H. and S.T.M.; methodology, S.T.M. T.N. and J.N.; investigation, S.T.M. T.N. J.N. I.A.R. Y.U. M.M. V.S.R. and S.M.C.S.L.; data curation, A.J.S. D.R.R. and J.R.H.; writing—original draft, E.H. S.T.M. J.N. A.J.S. and G.I.F.; writing—review & editing, T.N. R.F.N. and S.M.C.S.L.; funding acquisition, E.H.; resources, B.J.P. R.F.N. and S.M.C.S.L.; supervision, E.H. G.I.F. and S.M.C.S.L. S.T.M. and E.H. are the inventors on a PCT patent application titled “Methods of generating human cochlear hair cells.” The other authors declare no competing interests. We support inclusive, diverse, and equitable conduct of research. This work was supported by a Department of Defense US Army Medical Research and Materiel Command Congressionally Directed Medical Research Program grant W81XWH-18-1-0062 (E. H.), National Institutes of Health grants R01 DC013294 and R01 DC015788 (E.H.), K08 DC016034 (R.F.N.), JSPS KAKENHI grant JP21K09638 (T.N.), the Indiana University Health-Indiana University School of Medicine Strategic Research Initiative (E.H.), and the Ruth C. Holton Research Funds (E.H.). The authors would like to thank the Center for Contraception, Sexuality, and Abortion (CASA) in Leiden and The Hague for the collection of the human fetal material; B. Daye, T. van der Helm, F. Wei, J. Harkin, T. Saeki, M. Jafarkkhani, E. Longworth-Mills, P. McGuire, and H. Gao for their expert technical assistance; J. Zheng and K. Homma for antibodies against PRESTIN; and M. W. Kelley for his comments on the manuscript.

Funders	Funder number
Ruth C. Holton Research Funds
Indiana University Health-Indiana University School of Medicine Strategic Research Initiative
JSPS-NSF	21K09638
National Institutes of Health (NIH)	R01 DC015788, K08 DC016034, R01 DC013294
Department of Defense US Army Medical Research and Materiel Command Congressionally Directed Medical Research Program	W81XWH-18-1-0062

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

auditory
cochlea
hair cell
human
inner ear
organoid
pluripotent stem cell
scRNA-seq

ASJC Scopus subject areas

Molecular Medicine
Genetics
Cell Biology

Access to Document

10.1016/j.stem.2023.06.006

Cite this

Moore, S. T., Nakamura, T., Nie, J., Solivais, A. J., Aristizábal-Ramírez, I., Ueda, Y., Manikandan, M., Reddy, V. S., Romano, D. R., Hoffman, J. R., Perrin, B. J., Nelson, R. F., Frolenkov, G. I., Chuva de Sousa Lopes, S. M., & Hashino, E. (2023). Generating high-fidelity cochlear organoids from human pluripotent stem cells. Cell Stem Cell, 30(7), 950-961.e7. https://doi.org/10.1016/j.stem.2023.06.006

@article{469d0ead62534a069cfeca1c2a88b3ee,

title = "Generating high-fidelity cochlear organoids from human pluripotent stem cells",

abstract = "Mechanosensitive hair cells in the cochlea are responsible for hearing but are vulnerable to damage by genetic mutations and environmental insults. The paucity of human cochlear tissues makes it difficult to study cochlear hair cells. Organoids offer a compelling platform to study scarce tissues in vitro; however, derivation of cochlear cell types has proven non-trivial. Here, using 3D cultures of human pluripotent stem cells, we sought to replicate key differentiation cues of cochlear specification. We found that timed modulations of Sonic Hedgehog and WNT signaling promote ventral gene expression in otic progenitors. Ventralized otic progenitors subsequently give rise to elaborately patterned epithelia containing hair cells with morphology, marker expression, and functional properties consistent with both outer and inner hair cells in the cochlea. These results suggest that early morphogenic cues are sufficient to drive cochlear induction and establish an unprecedented system to model the human auditory organ.",

keywords = "auditory, cochlea, hair cell, human, inner ear, organoid, pluripotent stem cell, scRNA-seq",

author = "Moore, \{Stephen T.\} and Takashi Nakamura and Jing Nie and Solivais, \{Alexander J.\} and Isabel Aristiz{\'a}bal-Ram{\'i}rez and Yoshitomo Ueda and Mayakannan Manikandan and Reddy, \{V. Shweta\} and Romano, \{Daniel R.\} and Hoffman, \{John R.\} and Perrin, \{Benjamin J.\} and Nelson, \{Rick F.\} and Frolenkov, \{Gregory I.\} and \{Chuva de Sousa Lopes\}, \{Susana M.\} and Eri Hashino",

note = "Publisher Copyright: {\textcopyright} 2023 Elsevier Inc.",

year = "2023",

month = jul,

day = "6",

doi = "10.1016/j.stem.2023.06.006",

language = "English",

volume = "30",

pages = "950--961.e7",

number = "7",

}

Moore, ST, Nakamura, T, Nie, J, Solivais, AJ, Aristizábal-Ramírez, I, Ueda, Y, Manikandan, M, Reddy, VS, Romano, DR, Hoffman, JR, Perrin, BJ, Nelson, RF, Frolenkov, GI, Chuva de Sousa Lopes, SM & Hashino, E 2023, 'Generating high-fidelity cochlear organoids from human pluripotent stem cells', Cell Stem Cell, vol. 30, no. 7, pp. 950-961.e7. https://doi.org/10.1016/j.stem.2023.06.006

TY - JOUR

T1 - Generating high-fidelity cochlear organoids from human pluripotent stem cells

AU - Moore, Stephen T.

AU - Nakamura, Takashi

AU - Nie, Jing

AU - Solivais, Alexander J.

AU - Aristizábal-Ramírez, Isabel

AU - Ueda, Yoshitomo

AU - Manikandan, Mayakannan

AU - Reddy, V. Shweta

AU - Romano, Daniel R.

AU - Hoffman, John R.

AU - Perrin, Benjamin J.

AU - Nelson, Rick F.

AU - Frolenkov, Gregory I.

AU - Chuva de Sousa Lopes, Susana M.

AU - Hashino, Eri

PY - 2023/7/6

Y1 - 2023/7/6

N2 - Mechanosensitive hair cells in the cochlea are responsible for hearing but are vulnerable to damage by genetic mutations and environmental insults. The paucity of human cochlear tissues makes it difficult to study cochlear hair cells. Organoids offer a compelling platform to study scarce tissues in vitro; however, derivation of cochlear cell types has proven non-trivial. Here, using 3D cultures of human pluripotent stem cells, we sought to replicate key differentiation cues of cochlear specification. We found that timed modulations of Sonic Hedgehog and WNT signaling promote ventral gene expression in otic progenitors. Ventralized otic progenitors subsequently give rise to elaborately patterned epithelia containing hair cells with morphology, marker expression, and functional properties consistent with both outer and inner hair cells in the cochlea. These results suggest that early morphogenic cues are sufficient to drive cochlear induction and establish an unprecedented system to model the human auditory organ.

AB - Mechanosensitive hair cells in the cochlea are responsible for hearing but are vulnerable to damage by genetic mutations and environmental insults. The paucity of human cochlear tissues makes it difficult to study cochlear hair cells. Organoids offer a compelling platform to study scarce tissues in vitro; however, derivation of cochlear cell types has proven non-trivial. Here, using 3D cultures of human pluripotent stem cells, we sought to replicate key differentiation cues of cochlear specification. We found that timed modulations of Sonic Hedgehog and WNT signaling promote ventral gene expression in otic progenitors. Ventralized otic progenitors subsequently give rise to elaborately patterned epithelia containing hair cells with morphology, marker expression, and functional properties consistent with both outer and inner hair cells in the cochlea. These results suggest that early morphogenic cues are sufficient to drive cochlear induction and establish an unprecedented system to model the human auditory organ.

KW - auditory

KW - cochlea

KW - hair cell

KW - human

KW - inner ear

KW - organoid

KW - pluripotent stem cell

KW - scRNA-seq

UR - https://www.scopus.com/pages/publications/85163887587

UR - https://www.scopus.com/pages/publications/85163887587#tab=citedBy

U2 - 10.1016/j.stem.2023.06.006

DO - 10.1016/j.stem.2023.06.006

M3 - Article

C2 - 37419105

AN - SCOPUS:85163887587

SN - 1934-5909

VL - 30

SP - 950-961.e7

JO - Cell Stem Cell

JF - Cell Stem Cell

IS - 7

ER -

Generating high-fidelity cochlear organoids from human pluripotent stem cells

Abstract

Bibliographical note

Funding

UN SDGs

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this