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      Identification of recurrent USP48 and BRAF mutations in Cushing’s disease

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          Abstract

          Cushing’s disease results from corticotroph adenomas of the pituitary that hypersecrete adrenocorticotropin (ACTH), leading to excess glucocorticoid and hypercortisolism. Mutations of the deubiquitinase gene USP8 occur in 35–62% of corticotroph adenomas. However, the major driver mutations in USP8 wild-type tumors remain elusive. Here, we report recurrent mutations in the deubiquitinase gene USP48 (predominantly encoding p.M415I or p.M415V; 21/91 subjects) and BRAF (encoding p.V600E; 15/91 subjects) in corticotroph adenomas with wild-type USP8. Similar to USP8 mutants, both USP48 and BRAF mutants enhance the promoter activity and transcription of the gene encoding proopiomelanocortin (POMC), which is the precursor of ACTH, providing a potential mechanism for ACTH overproduction in corticotroph adenomas. Moreover, primary corticotroph tumor cells harboring BRAF V600E are sensitive to the BRAF inhibitor vemurafenib. Our study thus contributes to the understanding of the molecular mechanism of the pathogenesis of corticotroph adenoma and informs therapeutic targets for this disease.

          Abstract

          In this study the authors report USP48 and BRAF are frequently mutated in USP8 wild-type corticotroph adenomas, and cause Cushing’s disease mainly through promoting the promoter activity of POMC. Inhibition of BRAF may be a promising therapeutic strategy for the treatment of patients with BRAF-mutated corticotroph adenomas.

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          Most cited references27

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          The regulation of AP-1 activity by mitogen-activated protein kinases.

          M Karin (1995)
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            Morbidity and mortality in Cushing's disease: an epidemiological approach.

            Although Cushing's disease is a well documented clinical entity, there is no epidemiological information about it. The present study tries to obtain this information. Forty-nine patients affected by Cushing's disease living in Vizcaya (Spain) between 1975 and 1992 were considered for an epidemiological study. The prevalence of known cases at the end of 1992 was 39.1 per million inhabitants. The average incidence of newly diagnosed cases was 2.4 cases per million people per year. Cushing's disease was more frequent in women (n = 46) than in men (n = 3), with a ratio of 15:1. Diabetes mellitus and hypertension were observed in 38.7 and 55.1% of patients, respectively. Remission of Cushing's disease was achieved in 36 out of 41 patients (87.5%). In general, the mortality was higher than that expected for the control population (standardized mortality ratio, SMR 3.8, 95% confidence interval, CI 2.5-17.9, P < 0.03). Concerning the cause of death, the SMR of vascular disease was 5 (95% CI 3.4-48.6, P < 0.05). Higher age, persistence of hypertension and abnormalities of glucose metabolism after treatment, were independent predictors of mortality (multivariate analyses, P < 0.01). Prevalence of Cushing's disease was 39.1 cases/million inhabitants and average incidence was 2.4 cases/million per year. Mortality was elevated, due to vascular disease, associated with higher age, persistence of hypertension and impaired glucose metabolism.
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              Recurrent gain-of-function USP8 mutations in Cushing's disease

              Cushing's disease, also known as adrenocorticotropic hormone (ACTH)-secreting pituitary adenomas (PAs) that cause excess cortisol production, accounts for up to 85% of corticotrophin-dependent Cushing's syndrome cases. However, the genetic alterations in this disease are unclear. Here, we performed whole-exome sequencing of DNA derived from 12 ACTH-secreting PAs and matched blood samples, which revealed three types of somatic mutations in a candidate gene, USP8 (encoding ubiquitin-specific protease 8), exclusively in exon 14 in 8 of 12 ACTH-secreting PAs. We further evaluated somatic USP8 mutations in additional 258 PAs by Sanger sequencing. Targeted sequencing further identified a total of 17 types of USP8 variants in 67 of 108 ACTH-secreting PAs (62.04%). However, none of these mutations was detected in other types of PAs (n = 150). These mutations aggregate within the 14-3-3 binding motif of USP8 and disrupt the interaction between USP8 and 14-3-3 protein, resulting in an elevated capacity to protect EGFR from lysosomal degradation. Accordingly, PAs with mutated USP8 display a higher incidence of EGFR expression, elevated EGFR protein abundance and mRNA expression levels of POMC, which encodes the precursor of ACTH. PAs with mutated USP8 are significantly smaller in size and have higher ACTH production than wild-type PAs. In surgically resected primary USP8-mutated tumor cells, USP8 knockdown or blocking EGFR effectively attenuates ACTH secretion. Taken together, somatic gain-of-function USP8 mutations are common and contribute to ACTH overproduction in Cushing's disease. Inhibition of USP8 or EGFR is promising for treating USP8-mutated corticotrophin adenoma. Our study highlights the potentially functional mutated gene in Cushing's disease and provides insights into the therapeutics of this disease.
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                Author and article information

                Contributors
                zhaoyaohs@vip.sina.com
                huangcx@shsmu.edu.cn
                shiyongyong@gmail.com
                Journal
                Nat Commun
                Nat Commun
                Nature Communications
                Nature Publishing Group UK (London )
                2041-1723
                9 August 2018
                9 August 2018
                2018
                : 9
                : 3171
                Affiliations
                [1 ]ISNI 0000 0004 0368 8293, GRID grid.16821.3c, Shanghai Key Laboratory of Psychotic Disorders, , Shanghai Mental Health Center, Shanghai Jiao Tong University School of Medicine; Bio-X Institutes, Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders (Ministry of Education), and the Collaborative Innovation Center for Brain Science, Shanghai Jiao Tong University, ; Shanghai, 200030 China
                [2 ]ISNI 0000 0004 1798 5117, GRID grid.412528.8, Department of Otolaryngology Head and Neck Surgery & Center of Sleep Medicine, , Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, ; Shanghai, 200233 China
                [3 ]ISNI 0000 0004 0368 8293, GRID grid.16821.3c, Shanghai Institute of Immunology, Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, , Shanghai Jiao Tong University School of Medicine, ; Shanghai, 200025 China
                [4 ]ISNI 0000 0001 0125 2443, GRID grid.8547.e, Department of Neurosurgery, Huashan Hospital, Shanghai Medical College, , Fudan University, ; Shanghai, 200040 China
                [5 ]Shanghai Pituitary Tumor Center, Shanghai, 200040 China
                [6 ]ISNI 0000 0001 0125 2443, GRID grid.8547.e, Institute of Neurosurgery, , Fudan University, ; Shanghai, 200040 China
                [7 ]ISNI 0000000119573309, GRID grid.9227.e, CAS Key Laboratory of Systems Biology, CAS Center for Excellence in Molecular Cell Science, Innovation Center for Cell Signaling Network, Shanghai Institute of Biochemistry and Cell Biology, , Chinese Academy of Sciences, ; Shanghai, 200031 China
                [8 ]ISNI 0000 0004 1797 8419, GRID grid.410726.6, University of Chinese Academy of Sciences, ; Beijing, 100049 China
                [9 ]ISNI 0000000119573309, GRID grid.9227.e, Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, , Chinese Academy of Sciences, ; Shanghai, 201203 China
                [10 ]ISNI 0000 0001 0125 2443, GRID grid.8547.e, Department of Endocrinology, Huashan Hospital, Shanghai Medical College, , Fudan University, ; Shanghai, 200040 China
                [11 ]ISNI 0000 0001 0125 2443, GRID grid.8547.e, Department of Pathology, Huashan Hospital, Shanghai Medical College, , Fudan University, ; Shanghai, 200040 China
                [12 ]ISNI 0000 0001 0125 2443, GRID grid.8547.e, Department of Radiology, Huashan Hospital, Shanghai Medical College, , Fudan University, ; Shanghai, 200040 China
                [13 ]Shanghai Center for Bioinformation Technology (SCBIT), Shanghai Academy of Science and Technology, Shanghai, 201203 China
                [14 ]ISNI 0000 0001 0125 2443, GRID grid.8547.e, Department of Epidemiology, School of Public Health, , Fudan University, ; Shanghai, 200032 China
                [15 ]ISNI 0000 0001 0125 2443, GRID grid.8547.e, State Key Laboratory of Medical Neurobiology, Institute of Neurosurgery, Shanghai Medical College, , Fudan University, ; 200040 Shanghai, China
                [16 ]ISNI 0000000100241216, GRID grid.189509.c, Department of Pathology, Preston Robert Tisch Brain Tumor Center, , Duke University Medical Center, ; Durham, NC 27710 USA
                [17 ]ISNI 0000 0004 0368 8293, GRID grid.16821.3c, Institute of Neuropsychiatric Science and Systems Biological Medicine, , Shanghai Jiao Tong University, ; Shanghai, 200030 China
                [18 ]GRID grid.412631.3, Department of Psychiatry, , First Teaching Hospital of Xinjiang Medical University, ; Urumqi, Xinjiang 830054 China
                [19 ]ISNI 0000 0001 0455 0905, GRID grid.410645.2, The Affiliated Hospital of Qingdao University & The Biomedical Sciences Institute of Qingdao University (Qingdao Branch of SJTU Bio-X Institutes), , Qingdao University, ; Qingdao, Shandong 266003 China
                Author information
                http://orcid.org/0000-0003-0326-3124
                http://orcid.org/0000-0002-0174-9254
                http://orcid.org/0000-0003-3864-8382
                http://orcid.org/0000-0003-4374-1055
                http://orcid.org/0000-0003-1710-1505
                Article
                5275
                10.1038/s41467-018-05275-5
                6085354
                30093687
                e9330c42-c383-4d0c-9757-bcc760b0add4
                © The Author(s) 2018

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 26 July 2017
                : 13 June 2018
                Funding
                Funded by: the National Key Basic Research Program of China (973 Program)
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