Abstract 728: Tissue-specific tumorigenesis in multiple endocrine neoplasia type 1

Abstract

Abstract Multiple endocrine neoplasia type 1 (MEN1) is an inherited cancer syndrome caused by a germline mutation in the MEN1 gene. While a germline heterozygous mutation in the MEN1 gene predisposes tumor formation in specific tissues such as the endocrine pancreas, parathyroid glands and anterior pituitary, this tissue-specific tumorigenesis is not dependent on MEN1 mutations alone. In fact, a homozygous deletion of Men1 in mouse pancreatic exocrine tissue does not result in tumor formation, suggesting a tissue-specific mechanism. Loss of menin activates a menin-interacting protein retinoblastoma-binding-protein 5 (RBBP5). Since RBBP5 transcriptionally regulates DNA methyltransferase 1 (DNMT1), this causes global DNA hypermethylation and subsequent tumorigenesis in MEN1-target endocrine tissues. We hypothesize that while RBBP5 is ubiquitously expressed, it exclusively binds to the DNMT1 promoter in MEN1-target-tissues through its recruitment by tissue-specific factors. Using ChIP-PCR, we demonstrated that Rbbp5 is bound to the Dnmt1 promoter in MEN1-target-tissues, while not in non-target tissues. In the present study, we set out to identify and validate putative endocrine-specific factors involved in Rbbp5 binding to the Dnmt1 promoter. To investigate this tissue-specific occupancy of Rbbp5 at the Dnmt1 promoter, a ChIP-Seq analysis was carried out on DNA purified from pancreatic endocrine islet cells and pancreatic exocrine cells from wild type (WT) mice. These DNA samples were analyzed to identify differentially occupied DNA binding sites of menin and Rbbp5 by a high-throughput tissue-specific ChIP-seq assay. This analysis yielded a list of endocrine-specific genes regulated by both menin and Rbbp5. In order to further identify the tissue-specific factors that may alter menin-Rbbp5 binding to the Dnmt1 promoter, we performed tissue-specific RNA-Seq on endocrine and exocrine pancreas tissues from WT mice. Integrating tissue-specific ChIP-Seq and RNA-seq data, we identified 20 endocrine tissue-specific factors that may alter the interaction of the menin-Rbbp5 complex with the Dnmt1 promoter. By carrying out this unbiased genome-wide screen, we identified endocrine-specific candidate factors that may interact with the menin-Rbbp5 complex at the Dnmt1 promoter. After screening all candidates, the stand-out candidates were identified for further validation. Criteria to be met to establish stand-out candidates included: MEN1-target-tissue-specific expression through immunohistochemical staining and high mRNA expression by qRT-PCR analysis in WT mouse pancreatic islets on samples extracted by laser capture microscopy. In conclusion, we have identified candidates for Rbbp5 recruitment to the Dnmt1 promoter that must be tested further to determine their role in the observed tissue specificity of MEN1-related tumorigenesis. Citation Format: Juliet C. Gardiner. Tissue-specific tumorigenesis in multiple endocrine neoplasia type 1 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 728.