Norwegian Adenomyosis Study II: Gene Expression Profiling of Adenomyosis (NAPPED II)

Norwegian Adenomyosis Study: Pathophysiology, Peristalsis, Expression Profiling and Diagnosis, Part 2

Adenomyosis is characterized by the appearance of endometrial cells in the muscular layer of the uterus. It affects about 15-20% of the female population.

The symptoms of adenomyosis are heavy menstrual bleedings and painful menstruation (dysmenorrhea) and in addition chronic pelvic pain. Subfertility and infertility have been correlated with adenomyosis.

Parity, age and uterine abrasion increase the risk of adenomyosis. Hormonal factors such as local hyperestrogenism and elevated levels of prolactin (PRL) have been identified, but autoimmune and mechanical factors are also hypothesized.

Regarding treatment, the most effective measure is hysterectomy. As this is a very drastic measure in younger women, levonogestrel-releasing intrauterine devices, Gonadotropin releasing hormone (GnRH)-analogues, Danazol, uterine embolization and endometrial ablation have been tried, but studies are few in number, retrospective, and have small sample sizes.

Adenomyosis has so far not been subject to extensive research efforts. The pathogenesis of adenomyosis remains still unclear, there are not many satisfying treatment options and diagnostics include mostly magnetic resonance imaging (MRI) and histology.

The investigators designed a series of 3 studies with a broad approach in understanding adenomyosis. This is part 2.

In this study the investigators take both tissue samples and blood samples that will be investigated in order to understand the basic processes leading to adenomyosis.

Study Overview

• Status: Active, not recruiting
• Conditions: Adenomyosis
• Intervention / Treatment: Procedure: Myometrial biopsy; Procedure: endometrial biopsy

Detailed Description

Biopsy of focal adenomyosis of the myometrium:

This will be an extension of the NAPPED1-study. The investigators will perform ultrasound-guided transvaginal biopsies of the myometrium and collect venous blood samples.

As recent studies have suggested abnormalities in the regulation of specific genes in the development of adenomyosis, the investigators want to investigate differentially expressed genes in adenomyosis compared to eutopic endometrium. Using microarrays, the investigators can simultaneously screen differences in expression of thousands of genes in samples from the two groups. Profiling studies performed on endometrium of healthy individuals and of endometriosis show results that enable identification of biological processes and molecular mechanisms. Expression profiles can be used to identify molecular targets for therapeutic purposes. There are some very interesting studies that investigate drug treatment on a molecular level e.g. the effect of Danazol treatment on eutopic and ectopic endometrial tissue, but intramural adenomyosis has not been subject to gene profiling yet.

Tissue samples can be easily obtained after hysterectomy, but those samples will only represent older women, and cannot be used for consecutive monitoring of biochemical effects of treatment, as the uterus is removed. In order to investigate the pathophysiology of adenomyosis in younger women, and compare it to those in older individuals, as well as to evaluate effects of treatment, it is necessary to be able to obtain in-vivo samples.

The plan is therefore to take transvaginal, ultrasound-guided biopsy-samples from the uterus (myometrium) of all included patients at the beginning of their surgery, when the patient is under full anesthesia. The safety of comparable procedures has been shown in prior studies, but the investigators will further validate the safety of this method. The investigators believe that an in-vivo biopsy is a safe measure, and that representative samples of adenomyosis can be obtained.

The challenge with adenomyosis is that it is located intramyometrially. A transcervical biopsy will contaminate the sample with eutopic endometria, therefore it is most meaningful to take the biopsy transvaginally, but not through the cavity. After obtaining the biopsy sample, it will be examined histologically by imprint, to confirm if glandular cells are contained where expected. When good routines show a reliable level of specificity, the investigators will go further by selecting approximately 10 patients for gene profiling of adenomyosis. The investigators will also use endometrial biopsies (Pipelle) from those patients to see if there is a difference between intramural adenomyosis and their eutopic endometria. Most patients that undergo hysterectomy have taken a pipelle-biopsy routinely.

The levels of prolactin, Anti-müllerian hormone (AMH), Follicle stimulating hormone (FSH), luteinizing hormone (LH) and estrogen will also be taken, in order to determine if the severity of adenomyosis is only related to age as shown before, or also to hormonal activity.

• Study Type: Observational
• Enrollment (Anticipated): 80

Contacts and Locations

Study Locations

• Norway
  • Oslo, Norway, 0382
    • Gynecological department, Oslo University Hospital, Ullevål

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 30 years to 50 years (Adult)
• Accepts Healthy Volunteers: Yes
• Genders Eligible for Study: Female

• Sampling Method: Non-Probability Sample

Study Population

women referred to our clinic and volunteering to participate

Description

Inclusion Criteria:

Premenopausal women aged 30 - 50 years old scheduled for vaginal, abdominal or laparoscopic total hysterectomy one or more of the following clinical symptoms:

• bleeding disorders (menorrhagia, irregular bleeding, hypermenorrhoea),
• chronic pelvic pain,
• dysmenorrhoea,
• or dyspareunia junction zone definable

Exclusion Criteria:

• postmenopausal women,
• pregnancy
• gynecological cancer
• GnRH analog therapy or systemic hormone therapy in the last three months prior to hysterectomy
• junctional zone not identifiable

Study Plan

How is the study designed?

Design Details

• Observational Models: Case-Control
• Time Perspectives: Prospective

Number of groups / cohorts

2

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
Biopsy: adenomyosis; Myometrial biopsy Pipelle	Procedure: Myometrial biopsy; transvaginal ultrasound guided biopsy of the myometria; Procedure: endometrial biopsy; transcervical endometrial biopsy; Other Names: Pipelle
Biopsy: Healthy; Myometrial Biopsy Pipelle	Procedure: Myometrial biopsy; transvaginal ultrasound guided biopsy of the myometria; Procedure: endometrial biopsy; transcervical endometrial biopsy; Other Names: Pipelle

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Sensitivity and specificity of biopsies for adenomyotic tissue in percent (%)	Sensitivity and specificity of ultrasound guided myometrial biopsies for adenomyotic tissue	At time of hysterectomy

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
multiple comparison of gene expression, measured in fold	Expression profile for proliferative and invasive genes in adenomyotic tissue and surrounding stromal cells compared to healthy individuals, in fold	at time of biopsy taking
Frequency of complications related to biopsy taking in percent (%)		through 1 hour after biopsytaking
Serum levels of prolactin in mU/L	Serum levels of prolactin	at time of hysterectomy
Serum level of ER, in nmol/L	Serum level of estrogen.	at time of hysterectomy
Serum levels of FSH in U/L	Levels of follicle stimulating hormone.	at time of hysterectomy
Serum levels of AMH in pmol/L	Levels of Anti-Mullerian hormone.	at time of hysterectomy
Serum levels of LH in U/L	Levels of luteinizing hormone.	at time of hysterectomy
difference hormone serum-levels in fold	Comparison of serum-levels of PRL, FSH, AMH, LH and ER in patients with and without adenomyosis	at time of hysterectomy

Collaborators and Investigators

• Sponsor: Oslo University Hospital
• Collaborators: University of Oslo; Helse Sor-Ost
• Investigators: Study Chair: Erik Qvigstad, PhD, MD, Oslo University Hospital, Ullevål; Principal Investigator: Tina Tellum, MD, Oslo University Hospital

Study record dates

Study Major Dates

• Study Start (Actual): August 6, 2014
• Primary Completion (Anticipated): November 1, 2023
• Study Completion (Anticipated): December 1, 2024

Study Registration Dates

• First Submitted: July 17, 2014
• First Submitted That Met QC Criteria: July 22, 2014
• First Posted (Estimate): July 23, 2014

Study Record Updates

• Last Update Posted (Actual): March 23, 2023
• Last Update Submitted That Met QC Criteria: March 22, 2023
• Last Verified: March 1, 2023

More Information

Terms related to this study

• Keywords: Dysmenorrhea; Pelvic pain; Menorrhagia; expression profiling; uterine biopsies; serum hormones
• Additional Relevant MeSH Terms: Uterine Diseases; Adenomyosis
• Other Study ID Numbers: 2014/637b