Merlin-Deficient iPSCs Show Altered Pluripotency and Constitute a Potential in Vitro Model for NF2-Related Schwannomas

Núria Catasús; Miguel Torres-Martin; Alex Negro; Bernd Kuebler; Inma Rosas; Gemma Casals-Sendra; Helena Mazuelas; Francesc Roca-Ribas; Emilio Amilibia; Begoña Aran; Anna Veiga; Ángel Raya; Bernat Gel; Ignacio Blanco; Eduard Serra; Meritxell Carrió; Elisabeth Castellanos

doi:10.1101/2022.12.14.520389

Merlin-Deficient iPSCs Show Altered Pluripotency and Constitute a Potential in Vitro Model for NF2-Related Schwannomas

Núria Catasús, Miguel Torres-Martin, Alex Negro, Bernd Kuebler, Inma Rosas, Gemma Casals-Sendra, Helena Mazuelas, Francesc Roca-Ribas, Emilio Amilibia, Begoña Aran, Anna Veiga, Ángel Raya, Bernat Gel, Ignacio Blanco, Eduard Serra, Meritxell Carrió, Elisabeth Castellanos

Department of Surgery

Research output: Other contribution

Abstract

Background The appearance of bilateral vestibular schwannomas (VS) is one of the most characteristic features of NF2-related schwannomatosis (NF2-related SWN), an autosomal dominant syndrome that predisposes to the development of tumours of the nervous system. VS are caused by the bi-allelic inactivation of the NF2 gene in a cell of the Schwann cell lineage. Our current understanding of VS initiation and progression as well as the development of new effective therapies is hampered by the absence of human non-perishable cell-based models.

Principal Findings We generated and characterized induced pluripotent stem cell (iPSC) lines with single or bi-allelic inactivation of NF2 by combining the direct reprogramming of VS cells with the use of CRISPR/Cas9 editing. Despite the difficulty of maintaining merlin-deficient iPSCs, we were able to differentiate them into neural crest (NC) cells. At this stage, these cells showed spontaneous expression of the SC marker S100B and the impossibility of generating Schwann cells in 2D cultures. Nevertheless, by applying a 3D Schwann cell differentiation protocol, we successfully generated NF2(+/-) and NF2(-/-) spheroids homogeneously expressing classical markers of the NC-SC axis.

Conclusions Our results show a critical function of NF2 for both reprograming and maintaining a stable pluripotent state. In addition, merlin-deficient cultures also denoted an altered differentiation capacity of merlin-deficient cells towards the NC-SC axis, in the in vitro conditions used. Finally, the generated NF2(+/-) and NF2(-/-) spheroids show potential as a genuine in vitro model of NF2-related tumours.

Original language	Undefined/Unknown
DOIs	https://doi.org/10.1101/2022.12.14.520389
Publication status	Published - 14 Dec 2022

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Catasús, N., Torres-Martin, M., Negro, A., Kuebler, B., Rosas, I., Casals-Sendra, G., Mazuelas, H., Roca-Ribas, F., Amilibia, E., Aran, B., Veiga, A., Raya, Á., Gel, B., Blanco, I., Serra, E., Carrió, M., & Castellanos, E. (2022, Dec 14). Merlin-Deficient iPSCs Show Altered Pluripotency and Constitute a Potential in Vitro Model for NF2-Related Schwannomas. https://doi.org/10.1101/2022.12.14.520389

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title = "Merlin-Deficient iPSCs Show Altered Pluripotency and Constitute a Potential in Vitro Model for NF2-Related Schwannomas",

abstract = "Background The appearance of bilateral vestibular schwannomas (VS) is one of the most characteristic features of NF2-related schwannomatosis (NF2-related SWN), an autosomal dominant syndrome that predisposes to the development of tumours of the nervous system. VS are caused by the bi-allelic inactivation of the NF2 gene in a cell of the Schwann cell lineage. Our current understanding of VS initiation and progression as well as the development of new effective therapies is hampered by the absence of human non-perishable cell-based models.Principal Findings We generated and characterized induced pluripotent stem cell (iPSC) lines with single or bi-allelic inactivation of NF2 by combining the direct reprogramming of VS cells with the use of CRISPR/Cas9 editing. Despite the difficulty of maintaining merlin-deficient iPSCs, we were able to differentiate them into neural crest (NC) cells. At this stage, these cells showed spontaneous expression of the SC marker S100B and the impossibility of generating Schwann cells in 2D cultures. Nevertheless, by applying a 3D Schwann cell differentiation protocol, we successfully generated NF2(+/-) and NF2(-/-) spheroids homogeneously expressing classical markers of the NC-SC axis.Conclusions Our results show a critical function of NF2 for both reprograming and maintaining a stable pluripotent state. In addition, merlin-deficient cultures also denoted an altered differentiation capacity of merlin-deficient cells towards the NC-SC axis, in the in vitro conditions used. Finally, the generated NF2(+/-) and NF2(-/-) spheroids show potential as a genuine in vitro model of NF2-related tumours.",

author = "N{\'u}ria Catas{\'u}s and Miguel Torres-Martin and Alex Negro and Bernd Kuebler and Inma Rosas and Gemma Casals-Sendra and Helena Mazuelas and Francesc Roca-Ribas and Emilio Amilibia and Bego{\~n}a Aran and Anna Veiga and {\'A}ngel Raya and Bernat Gel and Ignacio Blanco and Eduard Serra and Meritxell Carri{\'o} and Elisabeth Castellanos",

year = "2022",

month = dec,

day = "14",

doi = "10.1101/2022.12.14.520389",

language = "Indefinido/desconocido",

type = "Other",

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Catasús, N, Torres-Martin, M, Negro, A, Kuebler, B, Rosas, I, Casals-Sendra, G, Mazuelas, H, Roca-Ribas, F, Amilibia, E, Aran, B, Veiga, A, Raya, Á, Gel, B, Blanco, I, Serra, E, Carrió, M & Castellanos, E 2022, Merlin-Deficient iPSCs Show Altered Pluripotency and Constitute a Potential in Vitro Model for NF2-Related Schwannomas.. https://doi.org/10.1101/2022.12.14.520389

TY - GEN

T1 - Merlin-Deficient iPSCs Show Altered Pluripotency and Constitute a Potential in Vitro Model for NF2-Related Schwannomas

AU - Catasús, Núria

AU - Torres-Martin, Miguel

AU - Negro, Alex

AU - Kuebler, Bernd

AU - Rosas, Inma

AU - Casals-Sendra, Gemma

AU - Mazuelas, Helena

AU - Roca-Ribas, Francesc

AU - Amilibia, Emilio

AU - Aran, Begoña

AU - Veiga, Anna

AU - Raya, Ángel

AU - Gel, Bernat

AU - Blanco, Ignacio

AU - Serra, Eduard

AU - Carrió, Meritxell

AU - Castellanos, Elisabeth

PY - 2022/12/14

Y1 - 2022/12/14

N2 - Background The appearance of bilateral vestibular schwannomas (VS) is one of the most characteristic features of NF2-related schwannomatosis (NF2-related SWN), an autosomal dominant syndrome that predisposes to the development of tumours of the nervous system. VS are caused by the bi-allelic inactivation of the NF2 gene in a cell of the Schwann cell lineage. Our current understanding of VS initiation and progression as well as the development of new effective therapies is hampered by the absence of human non-perishable cell-based models.Principal Findings We generated and characterized induced pluripotent stem cell (iPSC) lines with single or bi-allelic inactivation of NF2 by combining the direct reprogramming of VS cells with the use of CRISPR/Cas9 editing. Despite the difficulty of maintaining merlin-deficient iPSCs, we were able to differentiate them into neural crest (NC) cells. At this stage, these cells showed spontaneous expression of the SC marker S100B and the impossibility of generating Schwann cells in 2D cultures. Nevertheless, by applying a 3D Schwann cell differentiation protocol, we successfully generated NF2(+/-) and NF2(-/-) spheroids homogeneously expressing classical markers of the NC-SC axis.Conclusions Our results show a critical function of NF2 for both reprograming and maintaining a stable pluripotent state. In addition, merlin-deficient cultures also denoted an altered differentiation capacity of merlin-deficient cells towards the NC-SC axis, in the in vitro conditions used. Finally, the generated NF2(+/-) and NF2(-/-) spheroids show potential as a genuine in vitro model of NF2-related tumours.

AB - Background The appearance of bilateral vestibular schwannomas (VS) is one of the most characteristic features of NF2-related schwannomatosis (NF2-related SWN), an autosomal dominant syndrome that predisposes to the development of tumours of the nervous system. VS are caused by the bi-allelic inactivation of the NF2 gene in a cell of the Schwann cell lineage. Our current understanding of VS initiation and progression as well as the development of new effective therapies is hampered by the absence of human non-perishable cell-based models.Principal Findings We generated and characterized induced pluripotent stem cell (iPSC) lines with single or bi-allelic inactivation of NF2 by combining the direct reprogramming of VS cells with the use of CRISPR/Cas9 editing. Despite the difficulty of maintaining merlin-deficient iPSCs, we were able to differentiate them into neural crest (NC) cells. At this stage, these cells showed spontaneous expression of the SC marker S100B and the impossibility of generating Schwann cells in 2D cultures. Nevertheless, by applying a 3D Schwann cell differentiation protocol, we successfully generated NF2(+/-) and NF2(-/-) spheroids homogeneously expressing classical markers of the NC-SC axis.Conclusions Our results show a critical function of NF2 for both reprograming and maintaining a stable pluripotent state. In addition, merlin-deficient cultures also denoted an altered differentiation capacity of merlin-deficient cells towards the NC-SC axis, in the in vitro conditions used. Finally, the generated NF2(+/-) and NF2(-/-) spheroids show potential as a genuine in vitro model of NF2-related tumours.

U2 - 10.1101/2022.12.14.520389

DO - 10.1101/2022.12.14.520389

M3 - Otras contribuciones

ER -