Early Detection of Ovarian Cancer Using Uterine Lavage and Duplex - PowerPoint PPT Presentation

Early Detection of Ovarian Cancer Using Uterine Lavage and Duplex Sequencing GCIG Spring Meeting TRL Research Committee JUNE 2, 2017 Paul Speiser Medical University Vienna, Dept. Gynecologic Oncology Molecular Oncology Group GCIG Spring Meeting TRL Research Committee JUNE 2, 2017 / Paul Speiser Department Gynecologic Oncology, Molecular Oncology Group 1

Screening Recommendations US Preventative Services Task Force Grade A There is high certainty that the net benefit is substantial - recommended Grade: A Recommendation Screening cervical cancer ages 21 to 65 years with Pap smear every 3 years. Screening colorectal cancer : fecal occult blood testing, sigmoidoscopy, or colonoscopy, in adults, beginning at age 50 Innovative Concepts for Screening Detection of TP53 Mutations in Tampons of HGSC advanced stage patients 3 pos. out of 5 (60%) Obstet Gynecol 2014;124:881 – 5. DNA from Liquid Pap to detect Ovarian Cancer 22 Patients (4 early stage) 9 pos. (2eary stage) 41% www.ScienceTranslationalMedicine.org 9 January 2013 Vol 5 Issue 167 Uterine Lavage to detect Müllerian Duct Carcinomas 30 Patients (3 early stage) 24 pos. (3 early stage) 80% J Clin Oncol. 2015 Dec 20;33(36):4293-300. www.uspreventiveservicestaskforce.org

GCIG Spring Meeting TRL Research Committee JUNE 2, 2017 / Paul Speiser Department Gynecologic Oncology, Molecular Oncology Group 3

LUSTIC – L avage of the U terine cavity for diagnosis of S erous T ubal I ntraepithelial C arcinoma NCT02039388 Study aim: • Are exfoliated cells from STICs/occult carcinomas transported into the uterine cavity? • Is it possible to collect those cells via uterine lavage and detect them in the lavage fluid? → Earlier diagnosis of OC, or even its precursor lesions → Monitoring of HROC patients GCIG Spring Meeting TRL Research Committee JUNE 2, 2017 / Paul Speiser Department Gynecologic Oncology, Molecular Oncology Group 4

Study protocol: Histopath. Uterine examinatio lavage n GCIG Spring Meeting TRL Research Committee JUNE 2, 2017 / Paul Speiser Department Gynecologic Oncology, Molecular Oncology Group 5

Study protocol: STIC microdissection TP53 Sequencing Design a specific ddPCR assay TP53 mutation analysis of lavage sample GCIG Spring Meeting TRL Research Committee JUNE 2, 2017 / Paul Speiser Department Gynecologic Oncology, Molecular Oncology Group 6

LUSTIC # Patients Positive cases Klinikum Essen Mitte 34 ✓ Medical University of Graz ✓ 53 1 ✓ Tissue analysis  Foll. phase 1 Charles University Prag 59 ✓ Hamburg -Eppendorf 1 ✓ Lavage analysis 5 ✓ Universitätsklinikum  Shipment 4 Catholic University Leuven 27 ✓ ✓✓✓ Tissue analysis 1 ✓ Tissue analysis University College London 50 ✓ Trinity College Dublin 19 ✓ Radboud University Medical ✓ 5 Center ✓ Tissue analysis 1 Kopenhagen Rigshospitalet 11 ✓ Kepler Universitätsklinikum Linz ✓ 2 Brno Masaryk Memorial Cancer ✓ 7 Institute ✓ Lavage positive 2 Medical University of Vienna 36 ✓ Tissue analysis ✓ 11 308 = 3.6 % University Washington, Dept. Obst.&Gyn., Div. GYN ONC, Seminar, Mai 31 st 2017 Paul Speiser - Department Gynecologic Oncology, Molecular Oncology 7 Group

Applicability of the Lavage Concept for Screening Lavage IUD VAS Purposes local anesth. VAS minutes local anesth. VAS minutes interval 0 3 8 0 0 4 7 0 2.5 8 0 0 3.5 8 0 2.8 7 2.2 0 6.3 7 0 8.2 7 2.2 1 3.8 8 1 3.9 6 0 1 1 7 Proof of principle: 1 3.2 5 0 1 3.8 7 1 0 7 1 1 3 6 • Detection of early stage OC 1 3 5 0 1 2 7 1 1 7 0 1 1 7 • Detection of occult carcinoma 1 0 6 0 1 0 5 • 1 2 5 3 1 1 6 Detection of STIC 1 2.8 6 2.8 1 3.8 7 1 1 9 0 1 1 7 1 3 10 0 1 1 8 1 1.2 8 1 1 3 4 1 2 5 0 1 0 5 1 2 5 0 1 0.5 4 1 0 8 0 1 0 4 1 1 6 0 1 1.5 4 1 1 6 0 1 2 4 1 1 5 0 1 4 5 1 2 8 1 1 0.5 6 1 2.3 7 Median 1.8 6.7 1.7 6.04 GCIG Spring Meeting TRL Research Committee JUNE 2, 2017 / Paul Speiser Department Gynecologic Oncology, Molecular Oncology Group 8

GCIG Spring Meeting TRL Research Committee JUNE 2, 2017 / Paul Speiser Department Gynecologic Oncology, Molecular Oncology Group 9

Duplex Sequencing removes technological background Increase signal Decrease noise > 10,000-fold error reduction GCIG Spring Meeting TRL Research Committee JUNE 2, 2017 / Paul Speiser Department Gynecologic Oncology, Molecular Oncology Group 10

Tumor TP53 mutation is identified in 80% of ovarian cancer patients, >10-fold over biological background mutation Low-frequency TP53 mutations are prevalent in women with and without ovarian cancer: ‼ “ biological background ” GCIG Spring Meeting TRL Research Committee JUNE 2, 2017 / Paul Speiser Department Gynecologic Oncology, Molecular Oncology Group 11

Biological background TP53 mutations in uterine lavage from cases and controls increase with age GCIG Spring Meeting TRL Research Committee JUNE 2, 2017 / Paul Speiser Department Gynecologic Oncology, Molecular Oncology Group 12

Background mutations are not random Exonic background mutations by class Background mutations by pathogenicity Control cases Control cases Cancer cases Cancer cases Proportion of mutations Proportion of mutations Mutations are disproportionately Mutations are disproportionately pathogenic non-synonymous GCIG Spring Meeting TRL Research Committee JUNE 2, 2017 / Paul Speiser Department Gynecologic Oncology, Molecular Oncology Group

Background mutations are not random GCIG Spring Meeting TRL Research Committee JUNE 2, 2017 / Paul Speiser Department Gynecologic Oncology, Molecular Oncology Group

TP53 mutations in normal GU tissues Mutation load by tissue type Healthy 54 year old GCIG Spring Meeting TRL Research Committee JUNE 2, 2017 / Paul Speiser Department Gynecologic Oncology, Molecular Oncology Group

TP53 mutations in normal GU tissues Mutation load by tissue type Healthy 54 year old Average of 52-57 year olds GCIG Spring Meeting TRL Research Committee JUNE 2, 2017 / Paul Speiser Department Gynecologic Oncology, Molecular Oncology Group

Next steps... • Analysis of „ healthy tissue “ • SBIR Grant: Phase I Ideal sample preparation Validate sensitivity and reproducibility of high-througput DS Phase II TP53 Sequencing of uterine lavage samples of 100 average risk women with HGSC 100 average risk cancer-free controls 15 high-risk BRCA 1/2 mutation carriers with microscopic HGSC 100 high-risk BRCA 1/2 mutation carriers controls Morgenfortbildung 16.3.2017/ Elisabeth Maritschnegg Department of Obstetrics and Gynecology 17

The Early Detection Research Network (EDRN) June 7, 2017 Presentation to the Breast/GYN Collaborative Group LOVE Trial - Lavage of the uterine cavity and fallopian tubes for OVarian cancer Early detection Sensitivity & Specificity lavage concept in detecting STIC/occult EOC 2160 HBOC women, 6-monthly screen, 5%, 108 occult HGSC or STIC FDA Presubmission Meeting, Washington May 2016 Morgenfortbildung 16.3.2017/ Paul Speiser Department of Obstetrics and Gynecology 18

Acknowledgements University of Washington Larry Loeb Rosana Risques Katy Loubet-Senear Isaac Kinde, Yuxuan Wang, Lius Diaz, Kenneth Kinzler, Nickolas Loeb lab Papadopoulos, Bert Vogelstein Risques lab MUG, Universitätsklinik f. Frauenheilkunde u. Geburtshilfe , Gunda Kennedy lab Pristauz, Karl Tamussiono – Graz , Austria Klinikum Essen Mitte, Department Gynecology, Gynecologic Oncology, Florian Heitz, Andreas du Bois – Essen , Germany TwinStrand Biosciences Inc. Charité University - Campus Virchow Clinic, Department of Gynecology, Jalid Sehouli – Berlin , Germany Jesse Salk Charles University, Department of Obst.&Gyn., Gyn. Oncology Center, David Cibula – Prague , Czech Republic Charles University, Department of Obst.&Gyn., Dr. Jiri Bouda, Dr. Alena Bartakova – Pilsen , Czech Republic – 16 Samples Medical University of Vienna University Medical Center Hamburg-Eppendorf, Department of Gynecology, Sven Mahner – Hamburg , Germany Molecular Onvology Group Catholic University Leuven, Division Gynaecological Oncology, Ignace Vergote – Leuven , Belgium Robert Zeillinger The UK Familial Ovarian Cancer Screening Study (UK FOCSS), UCL, Martin Widschwendter – London , UK Elisabeth Maritschnegg Radboud Universiteit/Research Inst. Oncology, Univ. of Nijmegen Leon Massuger, – Nijmegen , Netherlands Trinity College, St James's Hospital, Department Gynaecologic Oncology, Noreen Gleeson – Dublin , Ireland University of Copenhagen, Rigshospitalet, Department of Gynaecology, Mikkel Rosendahl – Kopenhagen , Denmark