Early Detection of High Grade Ovarian Cancer Using Uterine Lavage EHUD Study and Duplex Sequencing

NCT04823871 | Completed

• 1 other identifier: EK 1612/2018
• Study Type: interventional
• Target: 406
• Locations: 4 countries
• Sites: 9

Brief Summary

In Phase I the sponsor will systematically test conditions for lavage filtration that increase tumor cell fraction without reducing tumor mutation yield. The Sponsor will also transition all lavages to luteal phase timing, when endometrial shedding is least. In Phase II the Sponsor will examine our data in context of clinical characteristics, particularly age, to develop a multivariate model that determines optimal mutant allele frequency (MAF) diagnostic threshold by patient. Furthermore, the sponsor will explore a highly innovative idea, entailing empirically determining each individual's background mutation load, agnostic of the aging or mutagenic exposures responsible, and using this as a personalized calibrator to determine optimal MAF diagnostic threshold.

Conditions

Ovarian Epithelial Cancer; Carcinoma in Situ; Ovarian Cancer

Keywords

Ovarian Epithelial Cancer f; Serous Intraepithelial Carcinoma (STIC); BRCA1 gene Mutation; BRCA2 gene Mutation; Neoplasms; Glandular and Epithelial; Ovarian Neoplasms; Ovarian cancer; Carcinoma in situ; Family history breast cancer and ovarian cancer

Outcome Measures

Primary Outcomes (7)

• Variant allele frequencies (VAF) for variants identified in both unfiltered and filtrate uterine lavage from the same patient.

  Preliminary testing with two samples indicated that filtration of UtLs to remove clusters of endometrial cells could increase sensitivity for tumor mutations several fold. This dataset will be expanded by analyzing the filtration effect in 10 ULs from patients with HGSC with known TP53 mutations. Each UtL will be divided in half: the first half will be filtered and the second will remain unfiltered. DNA will be extracted from the filtered, filtrate, and unfiltered fractions and analyzed by TP53KDS (total of 30 samples). For each UL, the DNA yield will be compared in each of the fractions and the MAF of the tumor mutation. It is anticipated that the filtered fraction will contain less DNA then the unfiltered fraction but the tumor mutation will be present at a higher frequency (i.e. more is gained by enrichment than is lost by reduction from less available DNA).

  Day 1

• SNP allele fraction (AF) comparison at different dilutions: expected AF vs. observed AF, AF variation among replicates (UtLs).

  SNP allele fraction (AF) will be compared at different dilutions to determine accuracy (expected AF vs. observed AF), precision (AF variation among replicates), and lowest limit of detection achievable.

  Day 1

• Total mutation frequency (UtLs)

  To asses the reproducibility in clinical use total mutation frequency will be assayed. The coefficient of variation among replicates will be calculated. Overall mutation frequency across TP53 in UtLs samples will be measured for both replicates of each patient. VAF will be approximated by number of non-reference duplex bases/total duplex bases sequenced. Confidence intervals will be computed using a binomial normal approximation.

  Day 1

• Mutant allele frequency (MAF) for all mutated positions, mutation spectrum (UtLs),

  A detailed mutation profile will be generated from the DS output files: mutant allele frequency (MAF) for all mutated positions, mutation spectrum, predicted pathogenicity to protein function, relationship to known hotspots, and overall mutation load (number of mutant nucleotides divided by the total number of nucleotides sequenced). Following this, samples will be unblinded for cases-controls status. TP53 MAF from DNA collected by UtLs will be used as predictor for differentiating between average risk patients (AIM I) with and without HGSC by logistic regression modelling. An analogous analysis will be applied to the group of high risk patients (AIM II) where presence and absence of STIC is defining the outcome variable. Cut off values for mutant allele frequency will be suggested in both cases and specificity and sensitivity will be estimated including appropriate confidence intervals.

  Day 1

• TP53 mutation frequencies in leukocytes

  The association between TP53 mutation frequencies in uterine lavage and leukocytes will be examined and will determine whether false negatives in Aim 1A correspond to cases with increased BB in leukocytes. In addition, TP53 mutation frequencies in leukocytes will be analyzed as a predictor of case control status, again using the leave out 10% procedure. This innovative calibration method will be compared with the simple ROC metrics achieved with the univariate model using a fixed mutation fraction as well as the adjusted multivariate model developed based on other patient characteristics such as age. As a scientific question unrelated to the present aims, it is worth investigating whether leukocyte TP53 mutation load will serve as an independent predictor of patient age, overall health or history of mutagenic exposures.

  Day 1

• Variant allele frequencies (VAF) for variants identified in uterine lavage ans pap smear from the same patient.

  Comparison of performance of UtL collection with that of Pap smear collection. It will be investigated if any additional power to discriminate cases from controls can be gained by combining information from Pap smears and ULs.

  Day 1

• dCT-PCR values from 96-plexed high-throughput MSREqPCR analysis

  Aiming to evaluate the potential for defining a DNA-Methylation signature for simple PCR testing, dCT-PCR values from 96-plexed high-throughput MSREqPCR analysis will be analysed by bioinformatics \& biostatistics. See Detailed description og the Aim III above for more details.

  Day 1

Study Arms (2)

High risk patients for breast and/or ovarian cancer

OTHER

Procedure/Surgery: Lavage of the Cavum uteri and proximal Fallopian tubes, performed in the luteal phase of the female cycle

Procedure: Speiser-catheter for uterine and tubal lavage

Suspected Ovarian Epithelial Cancer

OTHER

Procedure/Surgery: Lavage of the Cavum uteri and proximal Fallopian tubes, performed in the luteal phase of the female cycle

Procedure: Speiser-catheter for uterine and tubal lavage

Interventions

Speiser-catheter for uterine and tubal lavage PROCEDURE

Procedure/Surgery: Lavage of the Cavum uteri and proximal Fallopian tubes, performed in the luteal phase of the female cycle

High risk patients for breast and/or ovarian cancer; Suspected Ovarian Epithelial Cancer

Eligibility Criteria

• Age: 18 Years - 80 Years
• Gender Eligibility Details: female
• Healthy Volunteers: No
• Age Groups: Adult (18-64), Older Adult (65+)

You may qualify if:

• Written informed consent obtained
• Age ≥ 18 years and ≤ 80 years
• Women undergoing primary surgery for suspected High grade serous carcinoma (HGSC) If premenopausal in luteal phase (minimum 14 days after last day of menstrual period) or:
• Women with high risk for breast or ovarian cancer (HBOC) undergoing prophylactic resection of tubes with or without ovaries. If premenopausal in luteal phase except amenorrhea under hormonal contraception (incl. levonorgestrel -IUD)

You may not qualify if:

• Incapacitated women
• Pregnant women
• Prior hysterectomy
• Prior bilateral salpingectomy
• Prior tubal ligation
• First half of menstrual cycle
• Interval debulking
• Current cytotoxic chemotherapy

Sponsors & Collaborators

• Paul Speiser, Prof.MD,: lead

Study Sites (9)

Medizinische Universität Graz

Graz, Austria

Location

Medical University Innsbruck

Innsbruck, Austria

Location

Kepler Universitätsklinikum Linz

Linz, Austria

Location

Medical University Vienna, Dptm. of Obstetrics & Gynaecology

Vienna, 1090, Austria

Location

Masaryk Memorial Cancer Institute Brno

Brno, Czechia

Location

Charles University Prag

Prague, Czechia

Location

University Hospital Bonn

Bonn, Germany

Location

Gynaecological Clinic TU of Munich

München, Germany

Location

St. James Hospital

Dublin, Ireland

Location

MeSH Terms

Conditions

Carcinoma, Ovarian Epithelial; Carcinoma in Situ; Ovarian Neoplasms; Neoplasms

Condition Hierarchy (Ancestors)

Carcinoma; Neoplasms, Glandular and Epithelial; Neoplasms by Histologic Type; Endocrine Gland Neoplasms; Neoplasms by Site; Ovarian Diseases; Adnexal Diseases; Genital Diseases, Female; Female Urogenital Diseases; Female Urogenital Diseases and Pregnancy Complications; Urogenital Diseases; Genital Neoplasms, Female; Urogenital Neoplasms; Genital Diseases; Endocrine System Diseases; Gonadal Disorders

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: NON RANDOMIZED
• Masking: NONE
• Purpose: DIAGNOSTIC
• Intervention Model: PARALLEL
• Sponsor Type: OTHER
• Responsible Party: SPONSOR INVESTIGATOR
• PI Title: Clinical Professor

Model Details: Case control study in two patient population (AIM 1 in average risk population, AIM 2 in high risk population)

Study Record Dates

• First Submitted: February 4, 2020
• First Posted: April 1, 2021
• Study Start: November 1, 2018
• Primary Completion: June 30, 2022
• Study Completion: June 30, 2022
• Last Updated: September 2, 2022

Record last verified: 2022-09

Locations

AU (4); IE (1); DE (2); CZ (2)