CONTEXT: Germline loss-of-function CDKN1B gene variants cause the autosomal dominant syndrome of multiple endocrine neoplasia type 4 (MEN4). Even though pituitary neuroendocrine tumors are a well-known component of the syndrome, only 2 cases of Cushing's disease (CD) have so far been described in this setting. AIM: To screen a large cohort of CD patients for CDKN1B gene defects and to determine their functional effects. PATIENTS: We screened 211 CD patients (94.3% pediatric) by germline whole-exome sequencing (WES) only (n = 157), germline and tumor WES (n = 27), Sanger sequencing (n = 6), and/or germline copy number variant (CNV) analysis (n = 194). Sixty cases were previously unpublished. Variant segregation was investigated in the patients' families, and putative pathogenic variants were functionally characterized. RESULTS: Five variants of interest were found in 1 patient each: 1 truncating (p.Q107Rfs*12) and 4 nontruncating variants, including 3 missense changes affecting the CDKN1B protein scatter domain (p.I119T, p.E126Q, and p.D136G) and one 5' untranslated region (UTR) deletion (c.-29_-26delAGAG). No CNVs were found. All cases presented early (10.5 ± 1.3 years) and apparently sporadically. Aside from colon adenocarcinoma in 1 carrier, no additional neoplasms were detected in the probands or their families. In vitro assays demonstrated protein instability and disruption of the scatter domain of CDKN1B for all variants tested. CONCLUSIONS: Five patients with CD and germline CDKN1B variants of uncertain significance (n = 2) or pathogenic/likely pathogenic (n = 3) were identified, accounting for 2.6% of the patients screened. Our finding that germline CDKN1B loss-of-function may present as apparently sporadic, isolated pediatric CD has important implications for clinical screening and genetic counselling.

College of Medicine Sulaimani University Sulaimani Kurdistan Iraq

Department of Endocrinology Cochin Hospital Assistance Publique Hôpitaux de Paris Institut Cochin INSERM U1016 CNRS 8104 Paris Descartes University Paris France

Department of Laboratory Medicine and Pathology University of Minnesota Medical School Minneapolis Minnesota

Department of Pediatrics 2nd Faculty of Medicine Charles University and University Hospital Motol Prague Czech Republic

Epidemiology Branch Division of Intramural Population Health Research Eunice Kennedy Shriver National Institute of Child Health and Human Development Bethesda Maryland

Neurosurgery Unit for Pituitary and Inheritable Diseases National Institute of Neurological Disorders and Stroke Bethesda Maryland

Newborn Screening Program Wadsworth Center New York State Department of Health Albany New York

Section on Endocrinology and Genetics Eunice Kennedy Shriver National Institute of Child Health and Human Development Bethesda Maryland

Zobrazit více v PubMed

Pellegata NS, Quintanilla-Martinez L, Siggelkow H, et al. Germ-line mutations in PubMed PMC

Georgitsi M, Raitila A, Karhu A, et al. Germline PubMed

Agarwal SK, Mateo CM, Marx SJ. Rare germline mutations in cyclin-dependent kinase inhibitor genes in multiple endocrine neoplasia type 1 and related states. J Clin Endocrinol Metab. 2009;94(5):1826-1834. PubMed PMC

Molatore S, Marinoni I, Lee M, et al. A novel germline PubMed PMC

Belar O, De La Hoz C, Pérez-Nanclares G, Castaño L, Gaztambide S; Spanish MEN1 Group Novel mutations in PubMed

Malanga D, De Gisi S, Riccardi M, et al. Functional characterization of a rare germline mutation in the gene encoding the cyclin-dependent kinase inhibitor PubMed

Occhi G, Regazzo D, Trivellin G, et al. A novel mutation in the upstream open reading frame of the PubMed PMC

Pardi E, Mariotti S, Pellegata NS, et al. Functional characterization of a PubMed PMC

Sambugaro S, Di Ruvo M, Ambrosio MR, et al. Early onset acromegaly associated with a novel deletion in PubMed

Polyak K, Kato JY, Solomon MJ, et al. p27Kip1, a cyclin-Cdk inhibitor, links transforming growth factor-beta and contact inhibition to cell cycle arrest. Genes Dev. 1994;8(1):9-22. PubMed

Chu IM, Hengst L, Slingerland JM. The Cdk inhibitor p27 in human cancer: prognostic potential and relevance to anticancer therapy. Nat Rev Cancer. 2008;8(4):253-267. PubMed

Dahia PL, Aguiar RC, Honegger J, et al. Mutation and expression analysis of the PubMed

Lidhar K, Korbonits M, Jordan S, et al. Low expression of the cell cycle inhibitor p27Kip1 in normal corticotroph cells, corticotroph tumors, and malignant pituitary tumors. J Clin Endocrinol Metab. 1999;84(10):3823-3830. PubMed

Korbonits M, Chahal HS, Kaltsas G, et al. Expression of phosphorylated p27(Kip1) protein and Jun activation domain-binding protein 1 in human pituitary tumors. J Clin Endocrinol Metab. 2002;87(6):2635-2643. PubMed

Molatore S, Pellegata NS. The MENX syndrome and p27: relationships with multiple endocrine neoplasia. In: Martini L, ed. Prog Brain Res. Vol 182 Oxford, UK: Elsevier; 2010:295-320. PubMed

Frederiksen A, Rossing M, Hermann P, Ejersted C, Thakker RV, Frost M. Clinical features of multiple endocrine neoplasia type 4: novel pathogenic variant and review of published cases. J Clin Endocrinol Metab. 2019;104(9):3637-3646. PubMed PMC

Igreja S, Chahal HS, Akker SA, et al. Assessment of PubMed

Stratakis CA, Tichomirowa MA, Boikos S, et al. The role of germline PubMed PMC

Tichomirowa MA, Lee M, Barlier A, et al. Cyclin-dependent kinase inhibitor 1B ( PubMed

Hernández-Ramírez LC, Gam R, Valdés N, et al. Loss-of-function mutations in the CABLES1 gene are a novel cause of Cushing’s disease. Endocr Relat Cancer. 2017;24(8):379-392. PubMed PMC

Trivellin G, Correa RR, Batsis M, et al. Screening for GPR101 defects in pediatric pituitary corticotropinomas. Endocr Relat Cancer. 2016;23(5):357-365. PubMed PMC

Faucz FR, Tirosh A, Tatsi C, et al. Somatic USP8 gene mutations are a common cause of pediatric Cushing disease. J Clin Endocrinol Metab. 2017;102(8):2836-2843. PubMed PMC

Hernández-Ramírez LC, Tatsi C, Lodish MB, et al. Corticotropinoma as a component of carney complex. J Endocr Soc. 2017;1(7):918-925. PubMed PMC

Makri A, Bonella MB, Keil MF, et al. Children with MEN1 gene mutations may present first (and at a young age) with Cushing disease. Clin Endocrinol (Oxf). 2018;89(4):437-443. PubMed PMC

Cohen M, Persky R, Stegemann R, et al. Germline USP8 mutation associated with pediatric Cushing disease and other clinical features: a new syndrome. J Clin Endocrinol Metab. 2019;104(10):4676-4682. PubMed PMC

Tatsi C, Pankratz N, Lane J, et al. Large genomic aberrations in corticotropinomas are associated with greater aggressiveness. J Clin Endocrinol Metab. 2019;104(5):1792-1801. PubMed PMC

Wang K, Li M, Hakonarson H. ANNOVAR: functional annotation of genetic variants from high-throughput sequencing data. Nucleic Acids Res. 2010;38(16):e164. PubMed PMC

Liu X, Jian X, Boerwinkle E. dbNSFP: a lightweight database of human nonsynonymous SNPs and their functional predictions. Hum Mutat. 2011;32(8):894-899. PubMed PMC

Robinson JT, Thorvaldsdóttir H, Winckler W, et al. Integrative genomics viewer. Nat Biotechnol. 2011;29(1):24-26. PubMed PMC

Kopanos C, Tsiolkas V, Kouris A, et al. VarSome: the human genomic variant search engine. Bioinformatics. 2019;35(11):1978-1980. PubMed PMC

Karczewski KJ, Francioli LC, Tiao G, et al. Variation across 141 456 human exomes and genomes reveals the spectrum of loss-of-function intolerance across human protein-coding genes. bioRxiv 2019:531210.

Borsari S, Pardi E, Pellegata NS, et al. Loss of p27 expression is associated with MEN1 gene mutations in sporadic parathyroid adenomas. Endocrine. 2017;55(2):386-397. PubMed

Chang BL, Zheng SL, Isaacs SD, et al. A polymorphism in the CDKN1B gene is associated with increased risk of hereditary prostate cancer. Cancer Res. 2004;64(6):1997-1999. PubMed

Pappa V, Papageorgiou S, Papageorgiou E, et al. A novel p27 gene mutation in a case of unclassified myeloproliferative disorder. Leuk Res. 2005;29(2):229-231. PubMed

Lauter KB, Arnold A. Mutational analysis of CDKN1B, a candidate tumor-suppressor gene, in refractory secondary/tertiary hyperparathyroidism. Kidney Int. 2008;73(10):1137-1140. PubMed

Ojeda D, Lakhal B, Fonseca DJ, et al. Sequence analysis of the CDKN1B gene in patients with premature ovarian failure reveals a novel mutation potentially related to the phenotype. Fertil Steril 2011; 95:2658-2660 e2651. PubMed

Ruiz-Heredia Y, Sánchez-Vega B, Onecha E, et al. Mutational screening of newly diagnosed multiple myeloma patients by deep targeted sequencing. Haematologica. 2018;103(11):e544-e548. PubMed PMC

Gummlich L, Kähne T, Naumann M, Kilic E, Jung K, Dubiel W. New insights into the mechanism of COP9 Signalosome-Cullin-RING ubiquitin-ligase pathway deregulation in urological cancers. Int Rev Cell Mol Biol. 2016;323:181-229. PubMed

Daly AF, Jaffrain-Rea ML, Ciccarelli A, et al. Clinical characterization of familial isolated pituitary adenomas. J Clin Endocrinol Metab. 2006;91(9):3316-3323. PubMed

Alrezk R, Hannah-Shmouni F, Stratakis CA. MEN4 and CDKN1B mutations: the latest of the MEN syndromes. Endocr Relat Cancer. 2017;24(10):T195-T208. PubMed PMC

The Cancer Genome Atlas Network. Comprehensive molecular characterization of human colon and rectal cancer. Nature 2012; 487(7407):330-337. PubMed PMC

Esteban-Jurado C, Vila-Casadesús M, Garre P, et al. Whole-exome sequencing identifies rare pathogenic variants in new predisposition genes for familial colorectal cancer. Genet Med. 2015;17(2):131-142. PubMed PMC

Brandi ML, Gagel RF, Angeli A, et al. Guidelines for diagnosis and therapy of MEN type 1 and type 2. J Clin Endocrinol Metab. 2001;86(12):5658-5671. PubMed

McAllister SS, Becker-Hapak M, Pintucci G, Pagano M, Dowdy SF. Novel p27(kip1) C-terminal scatter domain mediates Rac-dependent cell migration independent of cell cycle arrest functions. Mol Cell Biol. 2003;23(1):216-228. PubMed PMC

Besson A, Gurian-West M, Schmidt A, Hall A, Roberts JM. p27Kip1 modulates cell migration through the regulation of RhoA activation. Genes Dev. 2004;18(8):862-876. PubMed PMC

Fujita N, Sato S, Katayama K, Tsuruo T. Akt-dependent phosphorylation of p27Kip1 promotes binding to 14-3-3 and cytoplasmic localization. J Biol Chem. 2002;277(32): 28706-28713. PubMed

Costa-Guda J, Marinoni I, Molatore S, Pellegata NS, Arnold A. Somatic mutation and germline sequence abnormalities in CDKN1B, encoding p27Kip1, in sporadic parathyroid adenomas. J Clin Endocrinol Metab. 2011;96(4):E701-E706. PubMed PMC

Elston MS, Meyer-Rochow GY, Dray M, Swarbrick M, Conaglen JV. Early onset primary hyperparathyroidism associated with a novel germline mutation in CDKN1B. Case Rep Endocrinol. 2015;2015:510985. PubMed PMC

Bugalho MJ, Domingues R. Uncommon association of cerebral meningioma, parathyroid adenoma and papillary thyroid carcinoma in a patient harbouring a rare germline variant in the CDKN1B gene. BMJ Case Rep 2016; 2016:bcr2015213934. PubMed PMC

Richards S, Aziz N, Bale S, et al. ; ACMG Laboratory Quality Assurance Committee. Standards and guidelines for the interpretation of sequence variants: a joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology. Genet Med. 2015;17(5):405-424. PubMed PMC

Russo AA, Jeffrey PD, Patten AK, Massagué J, Pavletich NP. Crystal structure of the p27Kip1 cyclin-dependent-kinase inhibitor bound to the cyclin A-Cdk2 complex. Nature. 1996;382(6589):325-331. PubMed

Connor MK, Kotchetkov R, Cariou S, et al. CRM1/Ran-mediated nuclear export of p27(Kip1) involves a nuclear export signal and links p27 export and proteolysis. Mol Biol Cell. 2003;14(1):201-213. PubMed PMC

Sekimoto T, Fukumoto M, Yoneda Y. 14-3-3 suppresses the nuclear localization of threonine 157-phosphorylated p27(Kip1). EMBO J. 2004;23(9):1934-1942. PubMed PMC

Al-Hebshi NN, Li S, Nasher AT, et al. Exome sequencing of oral squamous cell carcinoma in users of Arabian snuff reveals novel candidates for driver genes. Int J Cancer. 2016;139(2): 363-372. PubMed PMC

Martin D, Abba MC, Molinolo AA, et al. The head and neck cancer cell oncogenome: a platform for the development of precision molecular therapies. Oncotarget. 2014;5(19):8906-8923. PubMed PMC

Occhi G, Trivellin G, Ceccato F, et al. Prevalence of PubMed

Landa I, Montero-Conde C, Malanga D, et al. Allelic variant at -79 (C>T) in CDKN1B (p27Kip1) confers an increased risk of thyroid cancer and alters mRNA levels. Endocr Relat Cancer. 2010;17(2):317-328. PubMed

Capasso M, McDaniel LD, Cimmino F, et al. The functional variant rs34330 of CDKN1B is associated with risk of neuroblastoma. J Cell Mol Med. 2017;21(12):3224-3230. PubMed PMC

Ma H, Jin G, Hu Z, et al. Variant genotypes of CDKN1A and CDKN1B are associated with an increased risk of breast cancer in Chinese women. Int J Cancer. 2006;119(9):2173-2178. PubMed

Driver KE, Song H, Lesueur F, et al. ; Studies in Epidemiology and Risks of Cancer Heredity (SEARCH) Team Association of single-nucleotide polymorphisms in the cell cycle genes with breast cancer in the British population. Carcinogenesis. 2008;29(2):333-341. PubMed PMC

Canbay E, Eraltan IY, Cercel A, et al. CCND1 and CDKN1B polymorphisms and risk of breast cancer. Anticancer Res. 2010;30(7):3093-3098. PubMed

Wang W, Spitz MR, Yang H, Lu C, Stewart DJ, Wu X. Genetic variants in cell cycle control pathway confer susceptibility to lung cancer. Clin Cancer Res. 2007;13(19):5974-5981. PubMed

Cai H, Xiang YB, Qu S, et al. Association of genetic polymorphisms in cell-cycle control genes and susceptibility to endometrial cancer among Chinese women. Am J Epidemiol. 2011;173(11):1263-1271. PubMed PMC

Liu F, Wei YG, Luo LM, et al. Genetic variants of p21 and p27 and hepatocellular cancer risk in a Chinese Han population: a case-control study. Int J Cancer. 2013;132(9):2056-2064. PubMed

Cheng XK, Wang XJ, Li XD, Ren XQ. Genetic association between the cyclin-dependent kinase inhibitor gene p27/Kip1 polymorphism (rs34330) and cancer susceptibility: a meta-analysis. Sci Rep. 2017;7(1):44871. PubMed PMC

Sekiya T, Bronstein MD, Benfini K, et al. p27 variant and corticotropinoma susceptibility: a genetic and in vitro study. Endocr Relat Cancer. 2014;21(3):395-404. PubMed

Li G, Sturgis EM, Wang LE, et al. Association between the V109G polymorphism of the p27 gene and the risk and progression of oral squamous cell carcinoma. Clin Cancer Res. 2004;10(12 Pt 1):3996-4002. PubMed

Schöndorf T, Eisele L, Göhring UJ, et al. The V109G polymorphism of the p27 gene CDKN1B indicates a worse outcome in node-negative breast cancer patients. Tumour Biol. 2004;25(5-6):306-312. PubMed

Naidu R, Har YC, Taib NA. P27 V109G Polymorphism is associated with lymph node metastases but not with increased risk of breast cancer. J Exp Clin Cancer Res. 2007;26(1):133-140. PubMed

Camargo-Kosugi CM, da Silva ID, Sato H, et al. The V109G polymorphism in the p27 gene is associated with endometriosis. Eur J Obstet Gynecol Reprod Biol. 2009;145(2):180-183. PubMed

Mohamed FZ, Hussien YM, AlBakry MM, Mohamed RH, Said NM. Role of DNA repair and cell cycle control genes in ovarian cancer susceptibility. Mol Biol Rep. 2013;40(5):3757-3768. PubMed

Halkova T, Dvorakova S, Sykorova V, et al. Polymorphisms in selected DNA repair genes and cell cycle regulating genes involved in the risk of papillary thyroid carcinoma. Cancer Biomark. 2016;17(1):97-106. PubMed

Schumacher FR, Al Olama AA, Berndt SI, et al. ; Profile Study; Australian Prostate Cancer BioResource (APCB); IMPACT Study; Canary PASS Investigators; Breast and Prostate Cancer Cohort Consortium (BPC3); PRACTICAL (Prostate Cancer Association Group to Investigate Cancer-Associated Alterations in the Genome) Consortium; Cancer of the Prostate in Sweden (CAPS); Prostate Cancer Genome-wide Association Study of Uncommon Susceptibility Loci (PEGASUS); Genetic Associations and Mechanisms in Oncology (GAME-ON)/Elucidating Loci Involved in Prostate Cancer Susceptibility (ELLIPSE) Consortium. Association analyses of more than 140 000 men identify 63 new prostate cancer susceptibility loci. Nat Genet. 2018;50(7):928-936. PubMed PMC

Mahajan A, Taliun D, Thurner M, et al. Fine-mapping type 2 diabetes loci to single-variant resolution using high-density imputation and islet-specific epigenome maps. Nat Genet. 2018;50(11):1505-1513. PubMed PMC

Wei F, Xu J, Tang L, et al. p27(Kip1) V109G polymorphism and cancer risk: a systematic review and meta-analysis. Cancer Biother Radiopharm. 2012;27(10):665-671. PubMed PMC

Barbieri RB, Bufalo NE, Secolin R, et al. Polymorphisms of cell cycle control genes influence the development of sporadic medullary thyroid carcinoma. Eur J Endocrinol. 2014;171(6): 761-767. PubMed

Pasquali D, Circelli L, Faggiano A, et al. CDKN1B V109G polymorphism a new prognostic factor in sporadic medullary thyroid carcinoma. Eur J Endocrinol. 2011;164(3):397-404. PubMed

Kibel AS, Suarez BK, Belani J, et al. CDKN1A and CDKN1B polymorphisms and risk of advanced prostate carcinoma. Cancer Res. 2003;63(9):2033-2036. PubMed

Lu Y, Gao K, Zhang M, et al. Genetic Association Between CDKN1B rs2066827 Polymorphism and Susceptibility to Cancer. Medicine (Baltimore). 2015;94(46):e1217. PubMed PMC

Francisco G, Gonçalves FT, Luiz OC, et al. Polymorphisms in the p27kip-1 and prohibitin genes denote novel genes associated with melanoma risk in Brazil, a high ultraviolet index region. Melanoma Res. 2013;23(3):231-236. PubMed

Circelli L, Ramundo V, Marotta V, et al. ; Multidisciplinary Group for NeuroEndocrine Tumours of Naples Prognostic role of the CDNK1B V109G polymorphism in multiple endocrine neoplasia type 1. J Cell Mol Med. 2015;19(7): 1735-1741. PubMed PMC

Zobrazit více v PubMed

ClinicalTrials.gov
NCT00001595