title: Recurrent gain-of-function USP8 mutations in Cushing's disease.
abstract: 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.
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Zeng-Yi
authors: ZY
authors: Ma
authors: Bio-X Institutes, Ministry of Education Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Institute of Social Cognitive and Behavioral Sciences, Shanghai Jiao Tong University, Shanghai 200030, China.
authors: Zhi-Jian
authors: ZJ
authors: Song
authors: 1] Bio-X Institutes, Ministry of Education Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Institute of Social Cognitive and Behavioral Sciences, Shanghai Jiao Tong University, Shanghai 200030, China [2] Shanghai Key Laboratory of Psychotic Disorders, Shanghai Mental Health Center, Shanghai Jiao Tong University School of Medicine, Shanghai 200030, China.
authors: Jian-Hua
authors: JH
authors: Chen
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Yong-Fei
authors: YF
authors: Wang
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Shi-Qi
authors: SQ
authors: Li
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Liang-Fu
authors: LF
authors: Zhou
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Ying
authors: Y
authors: Mao
authors: Department of Endocrinology, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Yi-Ming
authors: YM
authors: Li
authors: State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors: Rong-Gui
authors: RG
authors: Hu
authors: Department of Endocrinology, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Zhao-Yun
authors: ZY
authors: Zhang
authors: Department of Endocrinology, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Hong-Ying
authors: HY
authors: Ye
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Ming
authors: M
authors: Shen
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Xue-Fei
authors: XF
authors: Shou
authors: Bio-X Institutes, Ministry of Education Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Institute of Social Cognitive and Behavioral Sciences, Shanghai Jiao Tong University, Shanghai 200030, China.
authors: Zhi-Qiang
authors: ZQ
authors: Li
authors: State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors: Hong
authors: H
authors: Peng
authors: Bio-X Institutes, Ministry of Education Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Institute of Social Cognitive and Behavioral Sciences, Shanghai Jiao Tong University, Shanghai 200030, China.
authors: Qing-Zhong
authors: QZ
authors: Wang
authors: Bio-X Institutes, Ministry of Education Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Institute of Social Cognitive and Behavioral Sciences, Shanghai Jiao Tong University, Shanghai 200030, China.
authors: Dai-Zhan
authors: DZ
authors: Zhou
authors: Bio-X Institutes, Ministry of Education Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Institute of Social Cognitive and Behavioral Sciences, Shanghai Jiao Tong University, Shanghai 200030, China.
authors: Xiao-Lan
authors: XL
authors: Qin
authors: Bio-X Institutes, Ministry of Education Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Institute of Social Cognitive and Behavioral Sciences, Shanghai Jiao Tong University, Shanghai 200030, China.
authors: Jue
authors: J
authors: Ji
authors: Bio-X Institutes, Ministry of Education Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Institute of Social Cognitive and Behavioral Sciences, Shanghai Jiao Tong University, Shanghai 200030, China.
authors: Jie
authors: J
authors: Zheng
authors: Department of Neuropathology, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Hong
authors: H
authors: Chen
authors: Department of Neuropathology, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Yin
authors: Y
authors: Wang
authors: Department of Radiology, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Dao-Ying
authors: DY
authors: Geng
authors: Department of Radiology, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Wei-Jun
authors: WJ
authors: Tang
authors: Department of Epidemiology, School of Public Health, Fudan University, Shanghai 200032, China.
authors: Chao-Wei
authors: CW
authors: Fu
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Zhi-Feng
authors: ZF
authors: Shi
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Yi-Chao
authors: YC
authors: Zhang
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Zhao
authors: Z
authors: Ye
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Wen-Qiang
authors: WQ
authors: He
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Qi-Lin
authors: QL
authors: Zhang
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Qi-Sheng
authors: QS
authors: Tang
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Rong
authors: R
authors: Xie
authors: Bio-X Institutes, Ministry of Education Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Institute of Social Cognitive and Behavioral Sciences, Shanghai Jiao Tong University, Shanghai 200030, China.
authors: Jia-Wei
authors: JW
authors: Shen
authors: Bio-X Institutes, Ministry of Education Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Institute of Social Cognitive and Behavioral Sciences, Shanghai Jiao Tong University, Shanghai 200030, China.
authors: Zu-Jia
authors: ZJ
authors: Wen
authors: Bio-X Institutes, Ministry of Education Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Institute of Social Cognitive and Behavioral Sciences, Shanghai Jiao Tong University, Shanghai 200030, China.
authors: Juan
authors: J
authors: Zhou
authors: Department of Neurosurgery, Shanghai 5th People's Hospital, Shanghai 200240, China.
authors: Tao
authors: T
authors: Wang
authors: Department of Neurosurgery, Shanghai 5th People's Hospital, Shanghai 200240, China.
authors: Shan
authors: S
authors: Huang
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Hui-Jia
authors: HJ
authors: Qiu
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Ni-Dan
authors: ND
authors: Qiao
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Yi
authors: Y
authors: Zhang
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Li
authors: L
authors: Pan
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Wei-Min
authors: WM
authors: Bao
authors: Department of Neurosurgery, Provincial hospital, Shandong University, Jinan, Shandong 250021, China.
authors: Ying-Chao
authors: YC
authors: Liu
authors: Shanghai Institute of Immunology & Department of Immunobiology and Microbiology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
authors: Chuan-Xin
authors: CX
authors: Huang
authors: 1] Bio-X Institutes, Ministry of Education Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Institute of Social Cognitive and Behavioral Sciences, Shanghai Jiao Tong University, Shanghai 200030, China [2] Shandong Provincial Key Laboratory of Metabolic Disease, the Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, China.
authors: Yong-Yong
authors: YY
authors: Shi
authors: Department of Neurosurgery, Shanghai Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
authors: Yao
authors: Y
authors: Zhao
mesh_terms: D048948:14-3-3 Proteins / Q000378:metabolism; D049913:ACTH-Secreting Pituitary Adenoma / Q000235:genetics / Q000628:therapy*; D000293:Adolescent; D000324:Adrenocorticotropic Hormone / Q000378:metabolism*; D000328:Adult; D001483:Base Sequence; D003480:Cushing Syndrome / Q000235:genetics*; D010450:Endopeptidases / Q000235:genetics* / Q000378:metabolism; D056827:Endosomal Sorting Complexes Required for Transport / Q000235:genetics* / Q000378:metabolism; D066246:ErbB Receptors / Q000037:antagonists & inhibitors* / Q000378:metabolism; D059472:Exome / Q000235:genetics; D005260:Female; D000077156:Gefitinib; D006801:Humans; D008297:Male; D008875:Middle Aged; D011333:Pro-Opiomelanocortin / Q000378:metabolism; D011485:Protein Binding / Q000235:genetics; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011799:Quinazolines / Q000494:pharmacology; D034622:RNA Interference; D034741:RNA, Small Interfering; D017422:Sequence Analysis, DNA; D043222:Ubiquitin Thiolesterase / Q000235:genetics* / Q000378:metabolism; D055815:Young Adult
chemical_list: D048948:14-3-3 Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D047428:Protein Kinase Inhibitors; D011799:Quinazolines; D034741:RNA, Small Interfering; D011333:Pro-Opiomelanocortin; D000324:Adrenocorticotropic Hormone; C512478:EGFR protein, human; D066246:ErbB Receptors; D010450:Endopeptidases; C541534:USP8 protein, human; D043222:Ubiquitin Thiolesterase; D000077156:Gefitinib