Modeling corticotroph deficiency with pituitary organoids supports the functional role of NFKB2 in human pituitary differentiation

Deficient Anterior pituitary with common Variable Immune Deficiency (DAVID) syndrome results from NFKB2 heterozygous mutations, causing adrenocorticotropic hormone deficiency (ACTHD) and primary hypogammaglobulinemia. While NFKB signaling plays a crucial role in the immune system, its connection to...

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Main Authors: Thi Thom Mac, Teddy Fauquier, Nicolas Jullien, Pauline Romanet, Heather Etchevers, Anne Barlier, Frederic Castinetti, Thierry Brue
Format: Article
Language:English
Published: eLife Sciences Publications Ltd 2024-11-01
Series:eLife
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Online Access:https://elifesciences.org/articles/90875
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author Thi Thom Mac
Teddy Fauquier
Nicolas Jullien
Pauline Romanet
Heather Etchevers
Anne Barlier
Frederic Castinetti
Thierry Brue
author_facet Thi Thom Mac
Teddy Fauquier
Nicolas Jullien
Pauline Romanet
Heather Etchevers
Anne Barlier
Frederic Castinetti
Thierry Brue
author_sort Thi Thom Mac
collection DOAJ
description Deficient Anterior pituitary with common Variable Immune Deficiency (DAVID) syndrome results from NFKB2 heterozygous mutations, causing adrenocorticotropic hormone deficiency (ACTHD) and primary hypogammaglobulinemia. While NFKB signaling plays a crucial role in the immune system, its connection to endocrine symptoms is unclear. We established a human disease model to investigate the role of NFKB2 in pituitary development by creating pituitary organoids from CRISPR/Cas9-edited human induced pluripotent stem cells (hiPSCs). Introducing homozygous TBX19K146R/K146R missense pathogenic variant in hiPSC, an allele found in congenital isolated ACTHD, led to a strong reduction of corticotrophs number in pituitary organoids. Then, we characterized the development of organoids harboring NFKB2D865G/D865G mutations found in DAVID patients. NFKB2D865G/D865G mutation acted at different levels of development with mutant organoids displaying changes in the expression of genes involved on pituitary progenitor generation (HESX1, PITX1, LHX3), hypothalamic secreted factors (BMP4, FGF8, FGF10), epithelial-to-mesenchymal transition, lineage precursors development (TBX19, POU1F1) and corticotrophs terminal differentiation (PCSK1, POMC), and showed drastic reduction in the number of corticotrophs. Our results provide strong evidence for the direct role of NFKB2 mutations in the endocrine phenotype observed in patients leading to a new classification of a NFKB2 variant of previously unknown clinical significance as pathogenic in pituitary development.
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spelling doaj-art-8c25604daae64966bbe1a5703a6ffb2a2024-12-17T13:41:09ZengeLife Sciences Publications LtdeLife2050-084X2024-11-011210.7554/eLife.90875Modeling corticotroph deficiency with pituitary organoids supports the functional role of NFKB2 in human pituitary differentiationThi Thom Mac0https://orcid.org/0000-0002-3746-2669Teddy Fauquier1https://orcid.org/0000-0003-1132-9402Nicolas Jullien2Pauline Romanet3Heather Etchevers4https://orcid.org/0000-0003-0201-3799Anne Barlier5Frederic Castinetti6Thierry Brue7https://orcid.org/0000-0001-8482-6691Aix-Marseille University, INSERM, UMR1251, Marseille Medical Genetics, Institut MarMaRa, Marseille, France; Hanoi Medical University Hospital, Hanoi, Viet NamAix-Marseille University, INSERM, UMR1251, Marseille Medical Genetics, Institut MarMaRa, Marseille, FranceAix-Marseille University, CNRS, UMR7051, Institut de NeuroPhysiopathologie, Marseille, FranceAix-Marseille University, INSERM, UMR1251, Marseille Medical Genetics, Institut MarMaRa, Marseille, France; Aix-Marseille University, APHM, INSERM, MMG, Laboratory of Molecular Biology, La Conception Hospital, Institut MarMaRa, Marseille, FranceAix-Marseille University, INSERM, UMR1251, Marseille Medical Genetics, Institut MarMaRa, Marseille, FranceAix-Marseille University, INSERM, UMR1251, Marseille Medical Genetics, Institut MarMaRa, Marseille, France; Aix-Marseille University, APHM, INSERM, MMG, Laboratory of Molecular Biology, La Conception Hospital, Institut MarMaRa, Marseille, France; Aix Marseille University, APHM, INSERM, MMG, Department of Endocrinology, La Conception Hospital, Institut MarMaRa, Marseille, FranceAix-Marseille University, INSERM, UMR1251, Marseille Medical Genetics, Institut MarMaRa, Marseille, France; Aix Marseille University, APHM, INSERM, MMG, Department of Endocrinology, La Conception Hospital, Institut MarMaRa, Marseille, FranceAix-Marseille University, INSERM, UMR1251, Marseille Medical Genetics, Institut MarMaRa, Marseille, France; Aix Marseille University, APHM, INSERM, MMG, Department of Endocrinology, La Conception Hospital, Institut MarMaRa, Marseille, FranceDeficient Anterior pituitary with common Variable Immune Deficiency (DAVID) syndrome results from NFKB2 heterozygous mutations, causing adrenocorticotropic hormone deficiency (ACTHD) and primary hypogammaglobulinemia. While NFKB signaling plays a crucial role in the immune system, its connection to endocrine symptoms is unclear. We established a human disease model to investigate the role of NFKB2 in pituitary development by creating pituitary organoids from CRISPR/Cas9-edited human induced pluripotent stem cells (hiPSCs). Introducing homozygous TBX19K146R/K146R missense pathogenic variant in hiPSC, an allele found in congenital isolated ACTHD, led to a strong reduction of corticotrophs number in pituitary organoids. Then, we characterized the development of organoids harboring NFKB2D865G/D865G mutations found in DAVID patients. NFKB2D865G/D865G mutation acted at different levels of development with mutant organoids displaying changes in the expression of genes involved on pituitary progenitor generation (HESX1, PITX1, LHX3), hypothalamic secreted factors (BMP4, FGF8, FGF10), epithelial-to-mesenchymal transition, lineage precursors development (TBX19, POU1F1) and corticotrophs terminal differentiation (PCSK1, POMC), and showed drastic reduction in the number of corticotrophs. Our results provide strong evidence for the direct role of NFKB2 mutations in the endocrine phenotype observed in patients leading to a new classification of a NFKB2 variant of previously unknown clinical significance as pathogenic in pituitary development.https://elifesciences.org/articles/90875pituitary developmentOrganoidscorticotroph deficitpituitary hormone deficitsACTH deficiencyDAVID syndrome
spellingShingle Thi Thom Mac
Teddy Fauquier
Nicolas Jullien
Pauline Romanet
Heather Etchevers
Anne Barlier
Frederic Castinetti
Thierry Brue
Modeling corticotroph deficiency with pituitary organoids supports the functional role of NFKB2 in human pituitary differentiation
eLife
pituitary development
Organoids
corticotroph deficit
pituitary hormone deficits
ACTH deficiency
DAVID syndrome
title Modeling corticotroph deficiency with pituitary organoids supports the functional role of NFKB2 in human pituitary differentiation
title_full Modeling corticotroph deficiency with pituitary organoids supports the functional role of NFKB2 in human pituitary differentiation
title_fullStr Modeling corticotroph deficiency with pituitary organoids supports the functional role of NFKB2 in human pituitary differentiation
title_full_unstemmed Modeling corticotroph deficiency with pituitary organoids supports the functional role of NFKB2 in human pituitary differentiation
title_short Modeling corticotroph deficiency with pituitary organoids supports the functional role of NFKB2 in human pituitary differentiation
title_sort modeling corticotroph deficiency with pituitary organoids supports the functional role of nfkb2 in human pituitary differentiation
topic pituitary development
Organoids
corticotroph deficit
pituitary hormone deficits
ACTH deficiency
DAVID syndrome
url https://elifesciences.org/articles/90875
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