PMID- 32780883 OWN - NLM STAT- MEDLINE DCOM- 20210728 LR - 20220531 IS - 1523-4681 (Electronic) IS - 0884-0431 (Linking) VI - 36 IP - 1 DP - 2021 Jan TI - Multiple Endocrine Neoplasia Type 1 (MEN1) 5'UTR Deletion, in MEN1 Family, Decreases Menin Expression. PG - 100-109 LID - 10.1002/jbmr.4156 [doi] AB - Multiple endocrine neoplasia type 1 (MEN1) is an autosomal dominant disorder characterized by the occurrence of parathyroid, pancreatic and pituitary tumors, and is due to mutations in the coding region of the MEN1 gene, which encodes menin. We investigated a family with identical twins that had MEN1, with different MEN1 tumors. DNA sequence analysis of the MEN1 coding region had not identified any abnormalities and we hypothesized that deletions and mutations involving the untranslated regions may be involved. Informed consent and venous blood samples were obtained from five family members. Sanger DNA sequencing and multiplex ligation-dependent probe amplification (MLPA) analyses were performed using leukocyte DNA. This revealed a heterozygous 596bp deletion (Delta596bp) between nucleotides -1087 and -492 upstream of the translation start site, located within the MEN1 5' untranslated region (UTR), and includes the core promoter and multiple cis-regulatory regions. To investigate the effects of this 5'UTR deletion on MEN1 promoter activity, we generated luciferase reporter constructs, containing either wild-type 842bp or mutant 246bp MEN1 promoter, and transfected them into human embryonic kidney HEK293 and pancreatic neuroendocrine tumor BON-1 cells. This revealed the Delta596bp mutation to result in significant reductions by 37-fold (p < 0.0001) and 16-fold (p < 0.0001) in luciferase expression in HEK293 and BON-1 cells, respectively, compared to wild-type. The effects of this 5'UTR deletion on MEN1 transcription and translation were assessed using qRT-PCR and Western blot analyses, respectively, of mRNA and protein lysates obtained from Epstein-Barr-virus transformed lymphoblastoid cells derived from affected and unaffected individuals. This demonstrated the Delta596bp mutation to result in significant reductions of 84% (p < 0.05) and 88% (p < 0.05) in MEN1 mRNA and menin protein, respectively, compared to unaffected individuals. Thus, our results report the first germline MEN1 5'UTR mutation and highlight the importance of investigating UTRs in MEN1 patients who do not have coding region mutations. (c) 2020 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR). CI - (c) 2020 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR). FAU - Kooblall, Kreepa G AU - Kooblall KG AUID- ORCID: 0000-0001-9547-2067 AD - Academic Endocrine Unit, Radcliffe Department of Medicine, University of Oxford, Oxford Centre for Diabetes, Endocrinology and Metabolism (OCDEM), Churchill Hospital, Oxford, UK. FAU - Boon, Hannah AU - Boon H AD - Oxford Medical Genetics Laboratory, Oxford University Hospitals NHS Trust, Oxford, UK. FAU - Cranston, Treena AU - Cranston T AD - Oxford Medical Genetics Laboratory, Oxford University Hospitals NHS Trust, Oxford, UK. FAU - Stevenson, Mark AU - Stevenson M AD - Academic Endocrine Unit, Radcliffe Department of Medicine, University of Oxford, Oxford Centre for Diabetes, Endocrinology and Metabolism (OCDEM), Churchill Hospital, Oxford, UK. FAU - Pagnamenta, Alistair T AU - Pagnamenta AT AD - Wellcome Trust Centre for Human Genetics, Oxford, UK. AD - Oxford NIHR Comprehensive Biomedical Research Centre, Oxford, UK. FAU - Rogers, Angela AU - Rogers A AD - Academic Endocrine Unit, Radcliffe Department of Medicine, University of Oxford, Oxford Centre for Diabetes, Endocrinology and Metabolism (OCDEM), Churchill Hospital, Oxford, UK. FAU - Grozinsky-Glasberg, Simona AU - Grozinsky-Glasberg S AD - Neuroendocrine Tumour Unit, ENETS Center of Excellence, Department of Endocrinology, Hadassah-Hebrew University Medical Centre, Jerusalem, Israel. FAU - Richardson, Tristan AU - Richardson T AD - Royal Bournemouth Hospital, Castle Lane East, Bournemouth, UK. FAU - Flanagan, Daniel Eh AU - Flanagan DE AD - Department of Endocrinology, Derriford Hospital, Plymouth, UK. CN - Genomics England Research Consortium AD - Genomics England Research Consortium, London, UK. AD - William Harvey Research Institute, Queen Mary University of London, London, UK. FAU - Taylor, Jenny C AU - Taylor JC AD - Wellcome Trust Centre for Human Genetics, Oxford, UK. AD - Oxford NIHR Comprehensive Biomedical Research Centre, Oxford, UK. FAU - Lines, Kate E AU - Lines KE AD - Academic Endocrine Unit, Radcliffe Department of Medicine, University of Oxford, Oxford Centre for Diabetes, Endocrinology and Metabolism (OCDEM), Churchill Hospital, Oxford, UK. FAU - Thakker, Rajesh V AU - Thakker RV AUID- ORCID: 0000-0002-1438-3220 AD - Academic Endocrine Unit, Radcliffe Department of Medicine, University of Oxford, Oxford Centre for Diabetes, Endocrinology and Metabolism (OCDEM), Churchill Hospital, Oxford, UK. LA - eng GR - G9825289/MRC_/Medical Research Council/United Kingdom GR - G1000467/MRC_/Medical Research Council/United Kingdom GR - DH_/Department of Health/United Kingdom GR - WT_/Wellcome Trust/United Kingdom GR - CRUK_/Cancer Research UK/United Kingdom PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20200915 PL - England TA - J Bone Miner Res JT - Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research JID - 8610640 RN - 0 (5' Untranslated Regions) RN - 0 (MEN1 protein, human) RN - 0 (Proto-Oncogene Proteins) SB - IM MH - 5' Untranslated Regions/genetics MH - Base Sequence MH - HEK293 Cells MH - Humans MH - *Multiple Endocrine Neoplasia Type 1/genetics MH - Proto-Oncogene Proteins MH - Sequence Analysis, DNA OTO - NOTNLM OT - GENETIC ANALYSIS OT - MONOZYGOTIC TWINS OT - NEUROENDOCRINE OT - PARATHYROID-RELATED DISORDERS OT - PROMOTER ACTIVITY EDAT- 2020/08/12 06:00 MHDA- 2021/07/29 06:00 CRDT- 2020/08/12 06:00 PHST- 2020/03/04 00:00 [received] PHST- 2020/07/22 00:00 [revised] PHST- 2020/08/02 00:00 [accepted] PHST- 2020/08/12 06:00 [pubmed] PHST- 2021/07/29 06:00 [medline] PHST- 2020/08/12 06:00 [entrez] AID - 10.1002/jbmr.4156 [doi] PST - ppublish SO - J Bone Miner Res. 2021 Jan;36(1):100-109. doi: 10.1002/jbmr.4156. Epub 2020 Sep 15.