TY - JOUR
T1 - Loss of KDM1A in GIP-dependent primary bilateral macronodular adrenal hyperplasia with Cushing's syndrome
T2 - a multicentre, retrospective, cohort study
AU - Chasseloup, Fanny
AU - Bourdeau, Isabelle
AU - Tabarin, Antoine
AU - Regazzo, Daniela
AU - Dumontet, Charles
AU - Ladurelle, Nataly
AU - Tosca, Lucie
AU - Amazit, Larbi
AU - Proust, Alexis
AU - Scharfmann, Raphael
AU - Mignot, Tiphaine
AU - Fiore, Frédéric
AU - Tsagarakis, Stylianos
AU - Vassiliadi, Dimitra
AU - Maiter, Dominique
AU - Young, Jacques
AU - Lecoq, Anne Lise
AU - Deméocq, Vianney
AU - Salenave, Sylvie
AU - Lefebvre, Hervé
AU - Cloix, Lucie
AU - Emy, Philippe
AU - Dessailloud, Rachel
AU - Vezzosi, Delphine
AU - Scaroni, Carla
AU - Barbot, Mattia
AU - de Herder, W.W.
AU - Pattou, François
AU - Tétreault, Martine
AU - Corbeil, Gilles
AU - Dupeux, Margot
AU - Lambert, Benoit
AU - Tachdjian, Gérard
AU - Guiochon-Mantel, Anne
AU - Beau, Isabelle
AU - Chanson, Philippe
AU - Viengchareun, Say
AU - Lacroix, André
AU - Bouligand, Jérôme
AU - Kamenický, Peter
N1 - Funding Information:
This study was funded by the Agence Nationale de la Recherche (ANR-18-CE14–0021 INTE-GRAL), Fondation du Grand défi Pierre Lavoie, and the French National Cancer Institute. FC received a Poste d'Accueil from INSERM and an Année Recherche fellowship from Assistance Publique–Hôpitaux de Paris. IBo received partial salary support from the Fonds de la recherche du Québec-Santé (FRQS). MT received a junior 1 salary award from the FRQS. We thank Céline Verstuyft (BB-0033–00089), Amandine Galioot, Sarah Cazenave, Emmanuelle Leteurtre, Marie Gaelle Barrande, Olivier Trassard, Nicolas Vendrisse, and Damien Schmitt for help with tissue samples. We thank Réseau de Recherche sur le cancer for sample collection at Centre hospitalier de l'Université de Montréal. We also thank Céline Piedvache and Kawther Nheri for help with statistical analyses and Séverine Pechberty for help with in-vitro experiments. RNA sequencing was done with support from the Paris Brain Institute iGenSeq and Data Analysis Core platforms.
Publisher Copyright:
© 2021 Elsevier Ltd
PY - 2021/12
Y1 - 2021/12
N2 - Background: GIP-dependent primary bilateral macronodular adrenal hyperplasia with Cushing's syndrome is caused by aberrant expression of the GIP receptor in adrenal lesions. The bilateral nature of this disease suggests germline genetic predisposition. We aimed to identify the genetic driver event responsible for GIP-dependent primary bilateral macronodular adrenal hyperplasia with Cushing's syndrome. Methods: We conducted a multicentre, retrospective, cohort study at endocrine hospitals and university hospitals in France, Canada, Italy, Greece, Belgium, and the Netherlands. We collected blood and adrenal samples from patients who had undergone unilateral or bilateral adrenalectomy for GIP-dependent primary bilateral macronodular adrenal hyperplasia with Cushing's syndrome. Adrenal samples from patients with primary bilateral macronodular adrenal hyperplasia who had undergone an adrenalectomy for overt or mild Cushing's syndrome without evidence of food-dependent cortisol production and those with GIP-dependent unilateral adrenocortical adenomas were used as control groups. We performed whole genome, whole exome, and targeted next generation sequencing, and copy number analyses of blood and adrenal DNA from patients with familial or sporadic disease. We performed RNA sequencing on adrenal samples and functional analyses of the identified genetic defect in the human adrenocortical cell line H295R. Findings: 17 patients with GIP-dependent primary bilateral macronodular adrenal hyperplasia with Cushing's syndrome were studied. The median age of patients was 43·3 (95% CI 38·8–47·8) years and most patients (15 [88%]) were women. We identified germline heterozygous pathogenic or most likely pathogenic variants in the KDM1A gene in all 17 patients. We also identified a recurrent deletion in the short p arm of chromosome 1 harboring the KDM1A locus in adrenal lesions of these patients. None of the 29 patients in the control groups had KDM1A germline or somatic alterations. Concomitant genetic inactivation of both KDM1A alleles resulted in loss of KDM1A expression in adrenal lesions. Global gene expression analysis showed GIP receptor upregulation with a log2 fold change of 7·99 (95% CI 7·34–8·66; p=4·4 × 10−125), and differential regulation of several other G protein-coupled receptors in GIP-dependent primary bilateral macronodular hyperplasia samples compared with control samples. In vitro pharmacological inhibition and inactivation of KDM1A by CRISPR-Cas9 genome editing resulted in an increase of GIP receptor transcripts and protein in human adrenocortical H295R cells. Interpretation: We propose that GIP-dependent primary bilateral macronodular adrenal hyperplasia with Cushing's syndrome results from a two-hit inactivation of KDM1A, consistent with the tumour suppressor gene model of tumorigenesis. Genetic testing and counselling should be offered to these patients and their relatives. Funding: Agence Nationale de la Recherche, Fondation du Grand défi Pierre Lavoie, and the French National Cancer Institute.
AB - Background: GIP-dependent primary bilateral macronodular adrenal hyperplasia with Cushing's syndrome is caused by aberrant expression of the GIP receptor in adrenal lesions. The bilateral nature of this disease suggests germline genetic predisposition. We aimed to identify the genetic driver event responsible for GIP-dependent primary bilateral macronodular adrenal hyperplasia with Cushing's syndrome. Methods: We conducted a multicentre, retrospective, cohort study at endocrine hospitals and university hospitals in France, Canada, Italy, Greece, Belgium, and the Netherlands. We collected blood and adrenal samples from patients who had undergone unilateral or bilateral adrenalectomy for GIP-dependent primary bilateral macronodular adrenal hyperplasia with Cushing's syndrome. Adrenal samples from patients with primary bilateral macronodular adrenal hyperplasia who had undergone an adrenalectomy for overt or mild Cushing's syndrome without evidence of food-dependent cortisol production and those with GIP-dependent unilateral adrenocortical adenomas were used as control groups. We performed whole genome, whole exome, and targeted next generation sequencing, and copy number analyses of blood and adrenal DNA from patients with familial or sporadic disease. We performed RNA sequencing on adrenal samples and functional analyses of the identified genetic defect in the human adrenocortical cell line H295R. Findings: 17 patients with GIP-dependent primary bilateral macronodular adrenal hyperplasia with Cushing's syndrome were studied. The median age of patients was 43·3 (95% CI 38·8–47·8) years and most patients (15 [88%]) were women. We identified germline heterozygous pathogenic or most likely pathogenic variants in the KDM1A gene in all 17 patients. We also identified a recurrent deletion in the short p arm of chromosome 1 harboring the KDM1A locus in adrenal lesions of these patients. None of the 29 patients in the control groups had KDM1A germline or somatic alterations. Concomitant genetic inactivation of both KDM1A alleles resulted in loss of KDM1A expression in adrenal lesions. Global gene expression analysis showed GIP receptor upregulation with a log2 fold change of 7·99 (95% CI 7·34–8·66; p=4·4 × 10−125), and differential regulation of several other G protein-coupled receptors in GIP-dependent primary bilateral macronodular hyperplasia samples compared with control samples. In vitro pharmacological inhibition and inactivation of KDM1A by CRISPR-Cas9 genome editing resulted in an increase of GIP receptor transcripts and protein in human adrenocortical H295R cells. Interpretation: We propose that GIP-dependent primary bilateral macronodular adrenal hyperplasia with Cushing's syndrome results from a two-hit inactivation of KDM1A, consistent with the tumour suppressor gene model of tumorigenesis. Genetic testing and counselling should be offered to these patients and their relatives. Funding: Agence Nationale de la Recherche, Fondation du Grand défi Pierre Lavoie, and the French National Cancer Institute.
UR - http://www.scopus.com/inward/record.url?scp=85119150855&partnerID=8YFLogxK
U2 - 10.1016/S2213-8587(21)00236-9
DO - 10.1016/S2213-8587(21)00236-9
M3 - Article
C2 - 34655521
AN - SCOPUS:85119150855
VL - 9
SP - 813
EP - 824
JO - The Lancet Diabetes and Endocrinology
JF - The Lancet Diabetes and Endocrinology
SN - 2213-8587
IS - 12
ER -