Oral Presentation HUPO 2019 - 18th Human Proteome Organization World Congress

Integrative -Omics Reveals Novel Targets Underlying the Pathomechanisms of Uterine Fibroids and Associated Heavy Menstrual Bleeding (#200)

Darragh P O'Brien 1 , Kavita S Subramaniam 2 , Thomas T Tapmeier 2 , Chen-Yi Wang 3 , Simon Davis 1 , Adam Cribbs 3 , Jessica Malzahn 3 , Thomas M Zollner 4 , Bianca De Leo 4 , Maik Oberdorf 4 , Joerg Muller 4 , Martin Fritsch 4 , Martin Philpott 3 , Krina T Zondervan 2 , Udo Oppermann 3 , Adrian Harris 5 , Christian M Becker 2 , Benedikt M Kessler 1
  1. Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom
  2. Endometriosis CaRe Centre, Nuffield Department of Women's & Reproductive Health, University of Oxford, Oxford, United Kingdom
  3. Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, University of Oxford, Oxford, United Kingdom
  4. Bayer AG, Berlin, Germany
  5. Department of Oncology, University of Oxford, Oxford, United Kingdom

Background: Uterine fibroids (UF’s) are benign tumours affecting up to 80% of women of reproductive age. UFs produce factors that induce angiogenesis with a highly irregular vasculature. Approximately 30% of fibroid patients suffer severe symptoms including pelvic pain and heavy menstrual bleeding (HMB). Although mutations in MED12 or HMGA2 account for the majority of UF occurrence, the processes by which these lead to UF’s and HMB remain poorly understood. Using an array of -Omics technologies, we undertake the first comprehensive integrative biology study of the pathomechanisms and vascularisation underlying UF formation and maintenance.

Methodologies: Fibroid, endometrium, myometrium, pseudocapsule, and healthy control samples (n = 130 tissues, across 45 donors) were obtained from patients undergoing surgery at the John Radcliffe Hospital, Oxford University Hospitals, UK. Gene and protein expression profiles between tissue types were investigated by RNA-Seq and quantitative proteomics. Spatial, cell-type resolved proteomic information was generated across tissues using Laser-Capture Microdissection Mass Spectrometry (LCM-MS) of uterine CD31+ cells. Candidate angiogenic factors were validated on FFPE tissues by immunohistochemistry and tested in vitro on human uterine microvascular endothelial cells. Their angiogenic potential was evaluated in proliferation and tube formation assays. Vascularised UF architecture was imaged by light sheet microscopy.

Findings: Integration of uterine tissue transcriptomics (>20,000 genes), proteomics (~5,500 proteins), and LCM-MS (~3,000 proteins), revealed altered biological pathways between different uterine tissues, e.g., Wnt signalling, which may serve as novel druggable targets for the treatment of UFs and HMB. Immunohistochemistry of UF vasculature confirmed the presence of candidate angiogenic factors, which were validated using tube length, endothelial mesh, and cell proliferation assays. Cleared whole mice uterus showed a distinct blood vessel pattern upon visualisation with 3D reconstruction imaging.

Concluding Statement: Using our multi -Omics approach, we have identified several exciting avenues to better understand the pathomechanisms of UFs and associated HMB.