Generation of hiPSC-derived low threshold mechanoreceptors containing axonal termini resembling bulbous sensory nerve endings and expressing Piezo1 and Piezo2

Shuyong Zhu; Nancy Stanslowsky; Jorge Fernández-Trillo; Tamrat M. Mamo; Pengfei Yu; Norman Kalmbach; Birgit Ritter; Reto Eggenschwiler; Werner J.D. Ouwendijk; David Mzinza; Likai Tan; Andreas Leffler; Michael Spohn; Richard J.P. Brown; Kai A. Kropp; Volkhard Kaever; Teng Cheong Ha; Pratibha Narayanan; Adam Grundhoff; Reinhold Förster; Axel Schambach; Georges M.G.M. Verjans; Manuela Schmidt; Andreas Kispert; Tobias Cantz; Ana Gomis; Florian Wegner; Abel Viejo-Borbolla

doi:10.1016/j.scr.2021.102535

Generation of hiPSC-derived low threshold mechanoreceptors containing axonal termini resembling bulbous sensory nerve endings and expressing Piezo1 and Piezo2

Shuyong Zhu^*
, Nancy Stanslowsky
, Jorge Fernández-Trillo
, Tamrat M. Mamo
, Pengfei Yu
, Norman Kalmbach
, Birgit Ritter
, Reto Eggenschwiler
, Werner J.D. Ouwendijk
, David Mzinza
, Likai Tan
, Andreas Leffler
, Michael Spohn
, Richard J.P. Brown
, Kai A. Kropp
, Volkhard Kaever
, Teng Cheong Ha
, Pratibha Narayanan
, Adam Grundhoff
, Reinhold Förster

Axel Schambach, Georges M.G.M. Verjans, Manuela Schmidt, Andreas Kispert, Tobias Cantz, Ana Gomis, Florian Wegner, Abel Viejo-Borbolla^*

^*Corresponding author for this work

Virology

Hannover Medical School
CSIC
Heinrich Pette Institute - Leibniz Institute for Experimental Virology
Paul-Ehrlich-Institut
Max Planck Institute of Experimental Medicine
Harvard Medical School

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

8 Citations (Scopus)

150 Downloads (Pure)

Abstract

Somatosensory low threshold mechanoreceptors (LTMRs) sense innocuous mechanical forces, largely through specialized axon termini termed sensory nerve endings, where the mechanotransduction process initiates upon activation of mechanotransducers. In humans, a subset of sensory nerve endings is enlarged, forming bulb-like expansions, termed bulbous nerve endings. There is no in vitro human model to study these neuronal endings. Piezo2 is the main mechanotransducer found in LTMRs. Recent evidence shows that Piezo1, the other mechanotransducer considered absent in dorsal root ganglia (DRG), is expressed at low level in somatosensory neurons. We established a differentiation protocol to generate, from iPSC-derived neuronal precursor cells, human LTMR recapitulating bulbous sensory nerve endings and heterogeneous expression of Piezo1 and Piezo2. The derived neurons express LTMR-specific genes, convert mechanical stimuli into electrical signals and have specialized axon termini that morphologically resemble bulbous nerve endings. Piezo2 is concentrated within these enlarged axon termini. Some derived neurons express low level Piezo1, and a subset co-express both channels. Thus, we generated a unique, iPSCs-derived human model that can be used to investigate the physiology of bulbous sensory nerve endings, and the role of Piezo1 and 2 during mechanosensation.

Original language	English
Article number	102535
Journal	Stem Cell Research
Volume	56
DOIs	https://doi.org/10.1016/j.scr.2021.102535
Publication status	Published - Oct 2021

Bibliographical note

Funding Information:
This work was supported by N-RENNT of the Ministry of Science and Culture of Lower Saxony to A.V.B., by a Marie Curie Career Integration Grant to A.V.B. (FP7-PEOPLE-2013-CIG, project number 631792, acronym INMA), by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy – EXC 2155 “RESIST” – Project ID 39087428, by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) - SFB-900 – 158989968 to R.F. (B1) and A.V.B. (B9), and by a Spanish Government project to A.G. (PID2019-108194RB-I00).

Funding Information:
We thank Ulrich Martin and Alexandra Haase (LEBAO, Hannover Medical School) for providing the iPSC line. We are thankful to Annette Garbe for performing LC-MS/MS quantification of neurotransmitters. We thank Martin Messerle (Hannover Medical School) for the ARPE-19 cells. We thank Emanuel Wyler (Max-Delbr?ck-Center for Molecular Medicine), Simon Krooss, Jens Bohne and Beate Sodeik (Hannover Medical School) for scientific advice. We thank Rudolf Bauerfeind, Michaela Kreienmeyer (Hannover Medical School) and Erik von Stedingk (BITPLANE), for technical support and Thomas Schulz (Hannover Medical School) for providing reagents. We are thankful to the Netherlands Brain Bank, Netherlands Institute for Neuroscience, Amsterdam, for providing the brain samples and bio samples. We are thankful to Miguel A. Valverde (Universitat Pompeu Fabra, Spain) for the pLKO1-Piezo2 shRNA and pLKO1-scramble shRNA plasmids. The raw and processed RNA-seq data has been submitted to Gene Expression Omnibus (GEO) as a GEO project and will be openly available.

Publisher Copyright:
© 2021 The Author(s)

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Generation of hiPSC-derived low threshold mechanoreceptors containing axonal termini resembling bulbous sensory nerve endings and expressing Piezo1 and Piezo2Final published version, 8.42 MBLicence: CC BY-NC-ND

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Zhu, S., Stanslowsky, N., Fernández-Trillo, J., Mamo, T. M., Yu, P., Kalmbach, N., Ritter, B., Eggenschwiler, R., Ouwendijk, W. J. D., Mzinza, D., Tan, L., Leffler, A., Spohn, M., Brown, R. J. P., Kropp, K. A., Kaever, V., Ha, T. C., Narayanan, P., Grundhoff, A., ... Viejo-Borbolla, A. (2021). Generation of hiPSC-derived low threshold mechanoreceptors containing axonal termini resembling bulbous sensory nerve endings and expressing Piezo1 and Piezo2. Stem Cell Research, 56, Article 102535. https://doi.org/10.1016/j.scr.2021.102535

@article{cd165912bc1448a2af24ba605935e545,

title = "Generation of hiPSC-derived low threshold mechanoreceptors containing axonal termini resembling bulbous sensory nerve endings and expressing Piezo1 and Piezo2",

abstract = "Somatosensory low threshold mechanoreceptors (LTMRs) sense innocuous mechanical forces, largely through specialized axon termini termed sensory nerve endings, where the mechanotransduction process initiates upon activation of mechanotransducers. In humans, a subset of sensory nerve endings is enlarged, forming bulb-like expansions, termed bulbous nerve endings. There is no in vitro human model to study these neuronal endings. Piezo2 is the main mechanotransducer found in LTMRs. Recent evidence shows that Piezo1, the other mechanotransducer considered absent in dorsal root ganglia (DRG), is expressed at low level in somatosensory neurons. We established a differentiation protocol to generate, from iPSC-derived neuronal precursor cells, human LTMR recapitulating bulbous sensory nerve endings and heterogeneous expression of Piezo1 and Piezo2. The derived neurons express LTMR-specific genes, convert mechanical stimuli into electrical signals and have specialized axon termini that morphologically resemble bulbous nerve endings. Piezo2 is concentrated within these enlarged axon termini. Some derived neurons express low level Piezo1, and a subset co-express both channels. Thus, we generated a unique, iPSCs-derived human model that can be used to investigate the physiology of bulbous sensory nerve endings, and the role of Piezo1 and 2 during mechanosensation.",

author = "Shuyong Zhu and Nancy Stanslowsky and Jorge Fern{\'a}ndez-Trillo and Mamo, \{Tamrat M.\} and Pengfei Yu and Norman Kalmbach and Birgit Ritter and Reto Eggenschwiler and Ouwendijk, \{Werner J.D.\} and David Mzinza and Likai Tan and Andreas Leffler and Michael Spohn and Brown, \{Richard J.P.\} and Kropp, \{Kai A.\} and Volkhard Kaever and Ha, \{Teng Cheong\} and Pratibha Narayanan and Adam Grundhoff and Reinhold F{\"o}rster and Axel Schambach and Verjans, \{Georges M.G.M.\} and Manuela Schmidt and Andreas Kispert and Tobias Cantz and Ana Gomis and Florian Wegner and Abel Viejo-Borbolla",

note = "Funding Information: This work was supported by N-RENNT of the Ministry of Science and Culture of Lower Saxony to A.V.B., by a Marie Curie Career Integration Grant to A.V.B. (FP7-PEOPLE-2013-CIG, project number 631792, acronym INMA), by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany{\textquoteright}s Excellence Strategy – EXC 2155 “RESIST” – Project ID 39087428, by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) - SFB-900 – 158989968 to R.F. (B1) and A.V.B. (B9), and by a Spanish Government project to A.G. (PID2019-108194RB-I00). Funding Information: We thank Ulrich Martin and Alexandra Haase (LEBAO, Hannover Medical School) for providing the iPSC line. We are thankful to Annette Garbe for performing LC-MS/MS quantification of neurotransmitters. We thank Martin Messerle (Hannover Medical School) for the ARPE-19 cells. We thank Emanuel Wyler (Max-Delbr?ck-Center for Molecular Medicine), Simon Krooss, Jens Bohne and Beate Sodeik (Hannover Medical School) for scientific advice. We thank Rudolf Bauerfeind, Michaela Kreienmeyer (Hannover Medical School) and Erik von Stedingk (BITPLANE), for technical support and Thomas Schulz (Hannover Medical School) for providing reagents. We are thankful to the Netherlands Brain Bank, Netherlands Institute for Neuroscience, Amsterdam, for providing the brain samples and bio samples. We are thankful to Miguel A. Valverde (Universitat Pompeu Fabra, Spain) for the pLKO1-Piezo2 shRNA and pLKO1-scramble shRNA plasmids. The raw and processed RNA-seq data has been submitted to Gene Expression Omnibus (GEO) as a GEO project and will be openly available. Publisher Copyright: {\textcopyright} 2021 The Author(s)",

year = "2021",

month = oct,

doi = "10.1016/j.scr.2021.102535",

language = "English",

volume = "56",

journal = "Stem Cell Research",

issn = "1873-5061",

publisher = "Elsevier",

}

Zhu, S, Stanslowsky, N, Fernández-Trillo, J, Mamo, TM, Yu, P, Kalmbach, N, Ritter, B, Eggenschwiler, R, Ouwendijk, WJD, Mzinza, D, Tan, L, Leffler, A, Spohn, M, Brown, RJP, Kropp, KA, Kaever, V, Ha, TC, Narayanan, P, Grundhoff, A, Förster, R, Schambach, A, Verjans, GMGM, Schmidt, M, Kispert, A, Cantz, T, Gomis, A, Wegner, F & Viejo-Borbolla, A 2021, 'Generation of hiPSC-derived low threshold mechanoreceptors containing axonal termini resembling bulbous sensory nerve endings and expressing Piezo1 and Piezo2', Stem Cell Research, vol. 56, 102535. https://doi.org/10.1016/j.scr.2021.102535

TY - JOUR

T1 - Generation of hiPSC-derived low threshold mechanoreceptors containing axonal termini resembling bulbous sensory nerve endings and expressing Piezo1 and Piezo2

AU - Zhu, Shuyong

AU - Stanslowsky, Nancy

AU - Fernández-Trillo, Jorge

AU - Mamo, Tamrat M.

AU - Yu, Pengfei

AU - Kalmbach, Norman

AU - Ritter, Birgit

AU - Eggenschwiler, Reto

AU - Ouwendijk, Werner J.D.

AU - Mzinza, David

AU - Tan, Likai

AU - Leffler, Andreas

AU - Spohn, Michael

AU - Brown, Richard J.P.

AU - Kropp, Kai A.

AU - Kaever, Volkhard

AU - Ha, Teng Cheong

AU - Narayanan, Pratibha

AU - Grundhoff, Adam

AU - Förster, Reinhold

AU - Schambach, Axel

AU - Verjans, Georges M.G.M.

AU - Schmidt, Manuela

AU - Kispert, Andreas

AU - Cantz, Tobias

AU - Gomis, Ana

AU - Wegner, Florian

AU - Viejo-Borbolla, Abel

N1 - Funding Information: This work was supported by N-RENNT of the Ministry of Science and Culture of Lower Saxony to A.V.B., by a Marie Curie Career Integration Grant to A.V.B. (FP7-PEOPLE-2013-CIG, project number 631792, acronym INMA), by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy – EXC 2155 “RESIST” – Project ID 39087428, by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) - SFB-900 – 158989968 to R.F. (B1) and A.V.B. (B9), and by a Spanish Government project to A.G. (PID2019-108194RB-I00). Funding Information: We thank Ulrich Martin and Alexandra Haase (LEBAO, Hannover Medical School) for providing the iPSC line. We are thankful to Annette Garbe for performing LC-MS/MS quantification of neurotransmitters. We thank Martin Messerle (Hannover Medical School) for the ARPE-19 cells. We thank Emanuel Wyler (Max-Delbr?ck-Center for Molecular Medicine), Simon Krooss, Jens Bohne and Beate Sodeik (Hannover Medical School) for scientific advice. We thank Rudolf Bauerfeind, Michaela Kreienmeyer (Hannover Medical School) and Erik von Stedingk (BITPLANE), for technical support and Thomas Schulz (Hannover Medical School) for providing reagents. We are thankful to the Netherlands Brain Bank, Netherlands Institute for Neuroscience, Amsterdam, for providing the brain samples and bio samples. We are thankful to Miguel A. Valverde (Universitat Pompeu Fabra, Spain) for the pLKO1-Piezo2 shRNA and pLKO1-scramble shRNA plasmids. The raw and processed RNA-seq data has been submitted to Gene Expression Omnibus (GEO) as a GEO project and will be openly available. Publisher Copyright: © 2021 The Author(s)

PY - 2021/10

Y1 - 2021/10

N2 - Somatosensory low threshold mechanoreceptors (LTMRs) sense innocuous mechanical forces, largely through specialized axon termini termed sensory nerve endings, where the mechanotransduction process initiates upon activation of mechanotransducers. In humans, a subset of sensory nerve endings is enlarged, forming bulb-like expansions, termed bulbous nerve endings. There is no in vitro human model to study these neuronal endings. Piezo2 is the main mechanotransducer found in LTMRs. Recent evidence shows that Piezo1, the other mechanotransducer considered absent in dorsal root ganglia (DRG), is expressed at low level in somatosensory neurons. We established a differentiation protocol to generate, from iPSC-derived neuronal precursor cells, human LTMR recapitulating bulbous sensory nerve endings and heterogeneous expression of Piezo1 and Piezo2. The derived neurons express LTMR-specific genes, convert mechanical stimuli into electrical signals and have specialized axon termini that morphologically resemble bulbous nerve endings. Piezo2 is concentrated within these enlarged axon termini. Some derived neurons express low level Piezo1, and a subset co-express both channels. Thus, we generated a unique, iPSCs-derived human model that can be used to investigate the physiology of bulbous sensory nerve endings, and the role of Piezo1 and 2 during mechanosensation.

AB - Somatosensory low threshold mechanoreceptors (LTMRs) sense innocuous mechanical forces, largely through specialized axon termini termed sensory nerve endings, where the mechanotransduction process initiates upon activation of mechanotransducers. In humans, a subset of sensory nerve endings is enlarged, forming bulb-like expansions, termed bulbous nerve endings. There is no in vitro human model to study these neuronal endings. Piezo2 is the main mechanotransducer found in LTMRs. Recent evidence shows that Piezo1, the other mechanotransducer considered absent in dorsal root ganglia (DRG), is expressed at low level in somatosensory neurons. We established a differentiation protocol to generate, from iPSC-derived neuronal precursor cells, human LTMR recapitulating bulbous sensory nerve endings and heterogeneous expression of Piezo1 and Piezo2. The derived neurons express LTMR-specific genes, convert mechanical stimuli into electrical signals and have specialized axon termini that morphologically resemble bulbous nerve endings. Piezo2 is concentrated within these enlarged axon termini. Some derived neurons express low level Piezo1, and a subset co-express both channels. Thus, we generated a unique, iPSCs-derived human model that can be used to investigate the physiology of bulbous sensory nerve endings, and the role of Piezo1 and 2 during mechanosensation.

UR - https://www.scopus.com/pages/publications/85116012975

U2 - 10.1016/j.scr.2021.102535

DO - 10.1016/j.scr.2021.102535

M3 - Article

AN - SCOPUS:85116012975

SN - 1873-5061

VL - 56

JO - Stem Cell Research

JF - Stem Cell Research

M1 - 102535

ER -

Generation of hiPSC-derived low threshold mechanoreceptors containing axonal termini resembling bulbous sensory nerve endings and expressing Piezo1 and Piezo2

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