Testis formation in XX individuals resulting from novel pathogenic variants in Wilms' tumor 1 (WT1) gene

Caroline Eozenou; Nitzan Gonen; Maria Sol Touzon; Anne Jorgensen; Svetlana A. Yatsenko; Leila Fusee; Alaa K. Kamel; Balazs Gellen; Gabriela Guercio; Priti Singh; Selma Witchel; Andrea J. Berman; Rana Mainpal; Mehdi Totonchi; Anahita Mohseni Meybodi; Masomeh Askari; Tiphanie Merel-Chali; Joelle Bignon-Topalovic; Roberta Migale; Mariana Costanzo; Roxana Marino; Pablo Ramirez; Natalia Perez Garrido; Esperanza Berensztein; Mona K. Mekkawy; John C. Schimenti; Rita Bertalan; Inas Mazen; Ken McElreavey; Alicia Belgorosky; Robin Lovell-Badge; Aleksandar Rajkovic; Anu Bashamboo

doi:10.1073/pnas.1921676117

Testis formation in XX individuals resulting from novel pathogenic variants in Wilms' tumor 1 (WT1) gene

Caroline Eozenou, Nitzan Gonen, Maria Sol Touzon, Anne Jorgensen, Svetlana A. Yatsenko, Leila Fusee, Alaa K. Kamel, Balazs Gellen, Gabriela Guercio, Priti Singh, Selma Witchel, Andrea J. Berman, Rana Mainpal, Mehdi Totonchi, Anahita Mohseni Meybodi, Masomeh Askari, Tiphanie Merel-Chali, Joelle Bignon-Topalovic, Roberta Migale, Mariana CostanzoRoxana Marino, Pablo Ramirez, Natalia Perez Garrido, Esperanza Berensztein, Mona K. Mekkawy, John C. Schimenti, Rita Bertalan, Inas Mazen, Ken McElreavey, Alicia Belgorosky, Robin Lovell-Badge, Aleksandar Rajkovic, Anu Bashamboo

The Mina and Everard Goodman Faculty of Life Sciences - at Bar-Ilan University

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

55 Scopus citations

Abstract

Sex determination in mammals is governed by antagonistic interactions of two genetic pathways, imbalance in which may lead to disorders/differences of sex development (DSD) in human. Among 46,XX individuals with testicular DSD (TDSD) or ovotesticular DSD (OTDSD), testicular tissue is present in the gonad. Although the testis-determining gene SRY is present in many cases, the etiology is unknown in most SRY-negative patients. We performed exome sequencing on 78 individuals with 46,XX TDSD/OTDSD of unknown genetic etiology and identified seven (8.97%) with heterozygous variants affecting the fourth zinc finger (ZF4) of Wilms' tumor 1 (WT1) (p.Ser478Thrfs∗17, p.Pro481Leufs∗15, p.Lys491Glu, p.Arg495Gln [x3], p.Arg495Gly). The variants were de novo in six families (P = 4.4 × 10^-6), and the incidence of WT1 variants in 46,XX DSD is enriched compared to control populations (P < 1.8 × 10^-4). The introduction of ZF4 mutants into a human granulosa cell line resulted in up-regulation of endogenous Sertoli cell transcripts and Wt1Arg495Gly/Arg495Gly XX mice display masculinization of the fetal gonads. The phenotype could be explained by the ability of the mutated proteins to physically interact with and sequester a key pro-ovary factor β-CATENIN, which may lead to up-regulation of testis-specific pathway. Our data show that unlike previous association of WT1 and 46,XY DSD, ZF4 variants of WT1 are a relatively common cause of 46,XX TDSD/OTDSD. This expands the spectrum of phenotypes associated with WT1 variants and shows that the WT1 protein affecting ZF4 can function as a protestis factor in an XX chromosomal context.

Original language	English
Pages (from-to)	13680-13688
Number of pages	9
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	117
Issue number	24
DOIs	https://doi.org/10.1073/pnas.1921676117
State	Published - 16 Jun 2020

Bibliographical note

Publisher Copyright:
© 2020 National Academy of Sciences. All rights reserved.

Funding

ACKNOWLEDGMENTS. A. Bashamboo is funded in part by a research grant from the European Society of Pediatric Endocrinology. K.M. and A. Bashamboo are funded by Agence Nationale de la Recherche ANR-10-LABX-73 REVIVE and Agence Nationale de la Recherche ANR-17-CE14-0038-01. This work is supported by the European Cooperation in Science and Technology (COST) Action DSDnet BM1303 (to A. Bashamboo and K.M.). N.G and R.L.-B. are funded by the Francis Crick Institute. The Francis Crick Institute receives its core funding from Cancer Research UK Grant FC001107, UK Medical Research Council Grant FC001107, Wellcome Grant FC001107, and by UK Medical Research Council Grant U117512772. M.S.T. and A. Belgorosky are supported by PIDC-20160028 Fondo Nacional de Ciencia y Tecnologia, Argentina, Grant PICT-2013-0181y PICT2016-0214; Agencia Nacional para Ciencia y Tecnologia, Argentina; and Consejo Nacional de Investigaciones Cientificas y Tecnologicas, Argentina. A.R. is funded by NIH Grants R01HD070647 and R21HD074278. A.J. is funded by a research grant from the Svend Andersen Foundation and the Danish Government’s support to Department of Growth and Reproduction for the International Center for Research and Research Training in Endocrine Disruption of Male Reproduction and Child Health (EDMaRC) programme. A.J.B. is supported by NIH Grant GM116889 and American Cancer Society Research Scholar Grant RSG-17-1.97-01-RMC. We are grateful to the Biological Research Facility, Genetic Modification Service, and Experimental Histopathology Facilities of the Francis Crick Institute. We acknowledge Dr. László Tiszlavicz (Pathological Department, University of Szeged, Hungary) for the histological examination of Patients 5a and 5b and Dr. Alejandro Suárez-Bonnet (Experimental Histopathology at Francis Crick Institute) for pathology report on mouse embryonic kidneys. A. Bashamboo is funded in part by a research grant from the European Society of Pediatric Endocrinology. K.M. and A. Bashamboo are funded by Agence Nationale de la Recherche ANR-10- LABX-73 REVIVE and Agence Nationale de la Recherche ANR-17-CE14- 0038-01. This work is supported by the European Cooperation in Science and Technology (COST) Action DSDnet BM1303 (to A. Bashamboo and K.M.). N.G and R.L.-B. are funded by the Francis Crick Institute. The Francis Crick Institute receives its core funding from Cancer Research UK Grant FC001107, UK Medical Research Council Grant FC001107, Wellcome Grant FC001107, and by UK Medical Research Council Grant U117512772. M.S.T. and A. Belgorosky are supported by PIDC-20160028 Fondo Nacional de Ciencia y Tecnologia, Argentina, Grant PICT-2013-0181y PICT2016-0214; Agencia Nacional para Ciencia y Tecnologia, Argentina; and Consejo Nacional de Investigaciones Cientificas y Tecnologicas, Argentina. A.R. is funded by NIH Grants R01HD070647 and R21HD074278. A.J. is funded by a research grant from the Svend Andersen Foundation and the Danish Government's support to Department of Growth and Reproduction for the International Center for Research and Research Training in Endocrine Disruption of Male Reproduction and Child Health (EDMaRC) programme. A.J.B. is supported by NIH Grant GM116889 and American Cancer Society Research Scholar Grant RSG-17-1.97-01-RMC. We are grateful to the Biological Research Facility, Genetic Modification Service, and Experimental Histopathology Facilities of the Francis Crick Institute. We acknowledge Dr. L?szl? Tiszlavicz (Pathological Department, University of Szeged, Hungary) for the histological examination of Patients 5a and 5b and Dr. Alejandro Su?rez-Bonnet (Experimental Histopathology at Francis Crick Institute) for pathology report on mouse embryonic kidneys.

Funders	Funder number
Agence Nationale de la Recherche	ANR-10-LABX-73
Agence Nationale de la Recherche ANR-10
Agence Nationale de la Recherche ANR-17-CE14-0038-01
Agencia Nacional para Ciencia y Tecnologia
American Cancer Society	RSG-17-1.97-01-RMC
COST	BM1303
Cancer Research UK	FC001107
Consejo Nacional de Investigaciones Cientificas y Tecnologicas
Department of Growth and Reproduction for the International Center for Research and Research Training in Endocrine Disruption of Male Reproduction and Child Health
EDMaRC
European Cooperation in Science and Technology
European Society of Pediatric Endocrinology
Fondo Nacional de Ciencia y Tecnologia	PICT-2013-0181y PICT2016-0214
Francis Crick Institute
Medical Research Council	U117512772
NIH	R21HD074278, R01HD070647
Svend Andersen Foundation
UK Medical Research Council
Wellcome
National Institute of General Medical Sciences	R01GM116889

Keywords

46,XX TDSD/OTDSD
Organogenesis
Sex determination
WT1
β-CATENIN

Access to Document

10.1073/pnas.1921676117

Cite this

Eozenou, C., Gonen, N., Touzon, M. S., Jorgensen, A., Yatsenko, S. A., Fusee, L., Kamel, A. K., Gellen, B., Guercio, G., Singh, P., Witchel, S., Berman, A. J., Mainpal, R., Totonchi, M., Meybodi, A. M., Askari, M., Merel-Chali, T., Bignon-Topalovic, J., Migale, R., ... Bashamboo, A. (2020). Testis formation in XX individuals resulting from novel pathogenic variants in Wilms' tumor 1 (WT1) gene. Proceedings of the National Academy of Sciences of the United States of America, 117(24), 13680-13688. https://doi.org/10.1073/pnas.1921676117

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abstract = "Sex determination in mammals is governed by antagonistic interactions of two genetic pathways, imbalance in which may lead to disorders/differences of sex development (DSD) in human. Among 46,XX individuals with testicular DSD (TDSD) or ovotesticular DSD (OTDSD), testicular tissue is present in the gonad. Although the testis-determining gene SRY is present in many cases, the etiology is unknown in most SRY-negative patients. We performed exome sequencing on 78 individuals with 46,XX TDSD/OTDSD of unknown genetic etiology and identified seven (8.97%) with heterozygous variants affecting the fourth zinc finger (ZF4) of Wilms' tumor 1 (WT1) (p.Ser478Thrfs∗17, p.Pro481Leufs∗15, p.Lys491Glu, p.Arg495Gln [x3], p.Arg495Gly). The variants were de novo in six families (P = 4.4 × 10-6), and the incidence of WT1 variants in 46,XX DSD is enriched compared to control populations (P < 1.8 × 10-4). The introduction of ZF4 mutants into a human granulosa cell line resulted in up-regulation of endogenous Sertoli cell transcripts and Wt1Arg495Gly/Arg495Gly XX mice display masculinization of the fetal gonads. The phenotype could be explained by the ability of the mutated proteins to physically interact with and sequester a key pro-ovary factor β-CATENIN, which may lead to up-regulation of testis-specific pathway. Our data show that unlike previous association of WT1 and 46,XY DSD, ZF4 variants of WT1 are a relatively common cause of 46,XX TDSD/OTDSD. This expands the spectrum of phenotypes associated with WT1 variants and shows that the WT1 protein affecting ZF4 can function as a protestis factor in an XX chromosomal context.",

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Eozenou, C, Gonen, N, Touzon, MS, Jorgensen, A, Yatsenko, SA, Fusee, L, Kamel, AK, Gellen, B, Guercio, G, Singh, P, Witchel, S, Berman, AJ, Mainpal, R, Totonchi, M, Meybodi, AM, Askari, M, Merel-Chali, T, Bignon-Topalovic, J, Migale, R, Costanzo, M, Marino, R, Ramirez, P, Garrido, NP, Berensztein, E, Mekkawy, MK, Schimenti, JC, Bertalan, R, Mazen, I, McElreavey, K, Belgorosky, A, Lovell-Badge, R, Rajkovic, A & Bashamboo, A 2020, 'Testis formation in XX individuals resulting from novel pathogenic variants in Wilms' tumor 1 (WT1) gene', Proceedings of the National Academy of Sciences of the United States of America, vol. 117, no. 24, pp. 13680-13688. https://doi.org/10.1073/pnas.1921676117

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T1 - Testis formation in XX individuals resulting from novel pathogenic variants in Wilms' tumor 1 (WT1) gene

AU - Eozenou, Caroline

AU - Gonen, Nitzan

AU - Touzon, Maria Sol

AU - Jorgensen, Anne

AU - Yatsenko, Svetlana A.

AU - Fusee, Leila

AU - Kamel, Alaa K.

AU - Gellen, Balazs

AU - Guercio, Gabriela

AU - Singh, Priti

AU - Witchel, Selma

AU - Berman, Andrea J.

AU - Mainpal, Rana

AU - Totonchi, Mehdi

AU - Meybodi, Anahita Mohseni

AU - Askari, Masomeh

AU - Merel-Chali, Tiphanie

AU - Bignon-Topalovic, Joelle

AU - Migale, Roberta

AU - Costanzo, Mariana

AU - Marino, Roxana

AU - Ramirez, Pablo

AU - Garrido, Natalia Perez

AU - Berensztein, Esperanza

AU - Mekkawy, Mona K.

AU - Schimenti, John C.

AU - Bertalan, Rita

AU - Mazen, Inas

AU - McElreavey, Ken

AU - Belgorosky, Alicia

AU - Lovell-Badge, Robin

AU - Rajkovic, Aleksandar

AU - Bashamboo, Anu

PY - 2020/6/16

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N2 - Sex determination in mammals is governed by antagonistic interactions of two genetic pathways, imbalance in which may lead to disorders/differences of sex development (DSD) in human. Among 46,XX individuals with testicular DSD (TDSD) or ovotesticular DSD (OTDSD), testicular tissue is present in the gonad. Although the testis-determining gene SRY is present in many cases, the etiology is unknown in most SRY-negative patients. We performed exome sequencing on 78 individuals with 46,XX TDSD/OTDSD of unknown genetic etiology and identified seven (8.97%) with heterozygous variants affecting the fourth zinc finger (ZF4) of Wilms' tumor 1 (WT1) (p.Ser478Thrfs∗17, p.Pro481Leufs∗15, p.Lys491Glu, p.Arg495Gln [x3], p.Arg495Gly). The variants were de novo in six families (P = 4.4 × 10-6), and the incidence of WT1 variants in 46,XX DSD is enriched compared to control populations (P < 1.8 × 10-4). The introduction of ZF4 mutants into a human granulosa cell line resulted in up-regulation of endogenous Sertoli cell transcripts and Wt1Arg495Gly/Arg495Gly XX mice display masculinization of the fetal gonads. The phenotype could be explained by the ability of the mutated proteins to physically interact with and sequester a key pro-ovary factor β-CATENIN, which may lead to up-regulation of testis-specific pathway. Our data show that unlike previous association of WT1 and 46,XY DSD, ZF4 variants of WT1 are a relatively common cause of 46,XX TDSD/OTDSD. This expands the spectrum of phenotypes associated with WT1 variants and shows that the WT1 protein affecting ZF4 can function as a protestis factor in an XX chromosomal context.

AB - Sex determination in mammals is governed by antagonistic interactions of two genetic pathways, imbalance in which may lead to disorders/differences of sex development (DSD) in human. Among 46,XX individuals with testicular DSD (TDSD) or ovotesticular DSD (OTDSD), testicular tissue is present in the gonad. Although the testis-determining gene SRY is present in many cases, the etiology is unknown in most SRY-negative patients. We performed exome sequencing on 78 individuals with 46,XX TDSD/OTDSD of unknown genetic etiology and identified seven (8.97%) with heterozygous variants affecting the fourth zinc finger (ZF4) of Wilms' tumor 1 (WT1) (p.Ser478Thrfs∗17, p.Pro481Leufs∗15, p.Lys491Glu, p.Arg495Gln [x3], p.Arg495Gly). The variants were de novo in six families (P = 4.4 × 10-6), and the incidence of WT1 variants in 46,XX DSD is enriched compared to control populations (P < 1.8 × 10-4). The introduction of ZF4 mutants into a human granulosa cell line resulted in up-regulation of endogenous Sertoli cell transcripts and Wt1Arg495Gly/Arg495Gly XX mice display masculinization of the fetal gonads. The phenotype could be explained by the ability of the mutated proteins to physically interact with and sequester a key pro-ovary factor β-CATENIN, which may lead to up-regulation of testis-specific pathway. Our data show that unlike previous association of WT1 and 46,XY DSD, ZF4 variants of WT1 are a relatively common cause of 46,XX TDSD/OTDSD. This expands the spectrum of phenotypes associated with WT1 variants and shows that the WT1 protein affecting ZF4 can function as a protestis factor in an XX chromosomal context.

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Testis formation in XX individuals resulting from novel pathogenic variants in Wilms' tumor 1 (WT1) gene

Abstract

Bibliographical note

Funding

Keywords

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