Regulation of Cervical Mucus Secretion

• ClinicalTrials.gov Identifier: NCT02969590
• Recruitment Status: Completed
• First Posted: November 21, 2016
• Results First Posted: October 18, 2019
• Last Update Posted: October 18, 2019
• Sponsor: Oregon Health and Science University
• Collaborator: Society of Family Planning
• Information provided by (Responsible Party): Leo Han, Oregon Health and Science University

Study Description

Brief Summary:

The purpose of this study is to help us determine how hormones impact the production of cervical mucus which is a fluid secreted by the opening of the uterus, also called the cervix. During a normal menstrual cycle, cervical mucus changes in response to two hormones that the ovaries make: estrogen and progesterone. Many of the birth control methods women use thicken mucus and make it less likely for sperm to enter the uterus. The goal of this study is to better understand the effects estrogen and progesterone have on the cervical mucus in the body. To do this, the investigators will be using a drug that suppresses natural hormone production for 3 months. Estrogen and progesterone will be artificially replaced to better understand their effects. The investigators will also be studying the effect of a common birth control pill that may work by thickening cervical mucus. Last, the investigators will study some of the cells in the cervix that produce mucus.

Condition or disease	Intervention/treatment	Phase
Fertility; Contraception	Drug: Leuprolide acetate; Drug: Estrogen and Progesterone Replacement; Drug: Progestin	Phase 4

Detailed Description:

This proposed study is to evaluate the temporal changes in human cervical mucus in response to progesterone and progestins as an initial step toward clarifying these relationships. Our hypothesis is that direct effects of progesterone on the endocervix, independent of estrogen withdrawal, cause contraceptive changes to cervical mucus. To study this, a randomized, prospective, crossover study will be conducted by examining cervical mucus changes in a small cohort of women in whom investigators will suppress circulating hormonal levels by administering a GnRH antagonist. Investigators will then artificially replace Estrogen and Progesterone in order to differentiate their effects on clinical and laboratory measurements of mucus quality. The investigators will be looking closely at the immediate changes in mucus when .35 mg of norethindrone, a marketed drug is administered in this experimental setting. Cervical cell samples will also be collected at various time points and perform RT-PCR to determine whether genes for membrane bound progesterone receptor are expressed and regulated by Estrogen and Progesterone.

Study Design

• Study Type: Interventional (Clinical Trial)
• Actual Enrollment: 5 participants
• Allocation: Randomized
• Intervention Model: Crossover Assignment
• Masking: Single (Investigator)
• Primary Purpose: Basic Science
• Official Title: Regulation of Cervical Mucus Secretion
• Actual Study Start Date: March 29, 2015
• Actual Primary Completion Date: August 23, 2017
• Actual Study Completion Date: August 23, 2017

Arms and Interventions

Arm	Intervention/treatment
No Intervention: No Intervention: Spontaneous Cycle (1 month); In order to qualify for the intervention phase, subjects needed to demonstrate favorable mucus at ovulation (Insler score of greater than or equal to 10 within 24h of an LH surge) and progesterone level in the luteal phase consistent with ovulation (a single P4 of greater than or equal to 3ng/ ml between days 18-35 of menstrual cycle).
Active Comparator: NET Arm - Norethindrone (4 months); This arm will receive Norethindrone (NET) first then experience estradiol withdrawal (E2WD). On the day prior to each admission, we instructed subjects to apply three fresh estradiol (E2) patches (0.3mg/day) to achieve mid-cycle level E2 effects. On the day of the first and third admission, subjects were randomized to the experimental condition, either: (1) oral progestin administration (e.g., a single dose of 0.35mg norethindrone) with continuation of the E2 patches; or (2) estradiol withdrawal (e.g., removal of the patches). Subject then received the converse intervention for admissions two and four. Subjects resumed treatment with the 0.1mg E2 patch until their next scheduled inpatient assessment. We required a minimum of five days at this level before repeating the dose escalation and the next evaluation.	Drug: Leuprolide acetate; Following this spontaneous cycle, participants received a single intramuscular injection of leuprolide acetate (11.25mg, LupronVR; Abbvie, Chicago, IL), a dose documented to suppress ovarian function for at least 3 months. Ovarian suppression is confirmed with serum E2 less than or equal to 35 pg/ml 21-28 day post-injection.; Other Name: Lupron; Drug: Estrogen and Progesterone Replacement; Replace hormones exogenously to create an artificial cycle; Other Names: Estradiol (vivelle-dot); Progesterone (prometrium, progestin); Drug: Progestin; Synthetic progestin to assess effects on cervical mucus.; Other Names: Norethindrone; NET
Active Comparator: E2WD Arm - Estradiol (4 months); This arm will experience estradiol withdrawal (E2WD) first and then receive Norethindrone (NET). On the day prior to each admission, we instructed subjects to apply three fresh estradiol (E2) patches (0.3mg/day) to achieve mid-cycle level E2 effects. On the day of the first and third admission, subjects were randomized to the experimental condition, either: (1) oral progestin administration (e.g., a single dose of 0.35mg norethindrone) with continuation of the E2 patches ; or (2) estradiol withdrawal (e.g., removal of the patches). Subject then received the converse intervention for admissions two and four. Subjects resumed treatment with the 0.1mg E2 patch until their next scheduled inpatient assessment. We required a minimum of five days at this level before repeating the dose escalation and the next evaluation.	Drug: Leuprolide acetate; Following this spontaneous cycle, participants received a single intramuscular injection of leuprolide acetate (11.25mg, LupronVR; Abbvie, Chicago, IL), a dose documented to suppress ovarian function for at least 3 months. Ovarian suppression is confirmed with serum E2 less than or equal to 35 pg/ml 21-28 day post-injection.; Other Name: Lupron; Drug: Estrogen and Progesterone Replacement; Replace hormones exogenously to create an artificial cycle; Other Names: Estradiol (vivelle-dot); Progesterone (prometrium, progestin); Drug: Progestin; Synthetic progestin to assess effects on cervical mucus.; Other Names: Norethindrone; NET

Outcome Measures

Primary Outcome Measures :

1. Median Cervical Mucus Score - Baseline [ Time Frame: Baseline ]
   Measurement of median cervical mucus scores at baseline. The current clinical standard for appraising cervical mucus is the cervical mucus score (ie. Insler score) that examines mucus based on 5 metrics including volume, spinnbarkeit (stretch), ferning, viscosity and cellularity on a 15-point scale. Per WHO guidelines, scores above 10 are considered "mucus favoring penetration" and scores below 10 are considered to be "unfavorable to penetration."
2. Sperm Penetration Scores [ Time Frame: Approximately one year ]
   Measuring sperm penetration scores in different hormonal conditions
3. Median Cervical Mucus Score - 2 Hour [ Time Frame: 2 hours ]
   Measurement of median cervical mucus scores 2 hours following norethindrone administration. The current clinical standard for appraising cervical mucus is the cervical mucus score (ie. Insler score) that examines mucus based on 5 metrics including volume, spinnbarkeit (stretch), ferning, viscosity and cellularity on a 15-point scale. Per WHO guidelines, scores above 10 are considered "mucus favoring penetration" and scores below 10 are considered to be "unfavorable to penetration."
4. Median Cervical Mucus Score - 6 Hour [ Time Frame: 6 hours ]
   Measurement of median cervical mucus scores 6 hours following norethindrone administration. The current clinical standard for appraising cervical mucus is the cervical mucus score (ie. Insler score) that examines mucus based on 5 metrics including volume, spinnbarkeit (stretch), ferning, viscosity and cellularity on a 15-point scale. Per WHO guidelines, scores above 10 are considered "mucus favoring penetration" and scores below 10 are considered to be "unfavorable to penetration."
5. Median Cervical Mucus Score - 24 Hour [ Time Frame: 24 hours ]
   Measurement of median cervical mucus scores 24 hours following norethindrone administration. The current clinical standard for appraising cervical mucus is the cervical mucus score (ie. Insler score) that examines mucus based on 5 metrics including volume, spinnbarkeit (stretch), ferning, viscosity and cellularity on a 15-point scale. Per WHO guidelines, scores above 10 are considered "mucus favoring penetration" and scores below 10 are considered to be "unfavorable to penetration."

Secondary Outcome Measures :

1. Change in PGRMC1 During Menstrual Cycle [ Time Frame: 1 month ]

   Measuring the mean transcript change of membrane bound progestin receptors (PGRMC1) from follicular phase to ovulation to luteal phase of spontaneous menstrual cycle.

   Total RNA (ribonucleic acid) was isolated from endocervical cell samples and analyzed for expression of PGRMC1 using real-time PCR (polymerase chain reaction) relative to levels of ribosomal (S10) RNA. An endocervical brush will be inserted into the os and then immediately rinsed into a special RNA preserving reagent. After total RNA is isolated and purified, it will be reverse transcribed into cDNA using primers.

   Ratio of PGRMC1 to 18s RNA

   Gene expression of membrane bound progesterone receptors in endocervical cells

Eligibility Criteria

• Ages Eligible for Study: 21 Years to 40 Years (Adult)
• Sexes Eligible for Study: Female
• Accepts Healthy Volunteers: Yes

Criteria

Inclusion Criteria:

• Normal menstrual cycles of 25-35 days in length for at least previous 3 cycles
• 21-40 years of age
• BMI >18, <30
• Serum P4 ≥ 3 ng/ml on single sample collected between days 18-25 of self-reported menstrual cycle
• Flexible schedule allowing morning blood draws on less than 48 hour notice
• In good general health
• Commit to remain on stable diet during study period (no changes to normal dietary habits)
• Commit to using non-hormonal contraceptive methods during study period except those prescribed in the experimental protocol
• No objections to taking study drugs
• No objections to refraining from intercourse the night before any sampling and willing to using condoms during vaginal intercourse.

Exclusion Criteria:

• Oral contraceptive use or other hormone supplement within the preceding 2 months
• Women with current cervical infection
• Evidence of abnormal cervical cytology
• Use of Paragard IUD for contraception
• Long-acting hormonal contraceptive use in the past 12 months (e.g., Depo-Provera®)
• Contraindications to study drugs
• Current or past pregnancy within the previous 6 months or currently trying to conceive
• Desiring to conceive in the next 8 months
• Breastfeeding in the past 2 months
• Diagnosed Diabetes or Metabolic Syndrome
• Current or previous use of cholesterol lowering drugs within the preceding 12 months
• Diagnosed Polycystic Ovary Syndrome
• History of, or self-reported, substance abuse
• Smoker
• Previous infertility treatment excluding male factor issues
• Use of an investigational drug within the past 2 months
• History of excisional or ablative treatment procedure on cervix (ie. LEEP, Cryotherapy, Cold Knife Cone)

Contacts and Locations

Locations

United States, Oregon

Oregon Health & Science University

Portland, Oregon, United States, 97239

Sponsors and Collaborators

Oregon Health and Science University

Society of Family Planning

Investigators

• Principal Investigator: Leo Han, MD; Oregon Health and Science University

  Study Documents (Full-Text)

Documents provided by Leo Han, Oregon Health and Science University:

Study Protocol and Statistical Analysis Plan  [PDF] November 12, 2018

More Information

Publications automatically indexed to this study by ClinicalTrials.gov Identifier (NCT Number):

Han L, Padua E, Edelman A, Jensen JT. Appraising cervical mucus: a new approach to evaluating contraceptives. Eur J Contracept Reprod Health Care. 2018 Feb;23(1):78-83. doi: 10.1080/13625187.2018.1437134. Epub 2018 Feb 19.

• Responsible Party: Leo Han, Instructor, Oregon Health and Science University
• ClinicalTrials.gov Identifier: NCT02969590
• Other Study ID Numbers: OHSU IRB 11532
• First Posted: November 21, 2016
• Results First Posted: October 18, 2019
• Last Update Posted: October 18, 2019
• Last Verified: October 2019

Keywords provided by Leo Han, Oregon Health and Science University:

Contraception; Mucus; Cervix; progestin; hormones

Additional relevant MeSH terms:

Leuprolide; Norethindrone; Estradiol; Estrogens; Progesterone; Progestins; Hormones; Hormones, Hormone Substitutes, and Hormone Antagonists; Physiological Effects of Drugs; Contraceptive Agents; Reproductive Control Agents; Contraceptive Agents, Female; Fertility Agents, Female; Fertility Agents; Antineoplastic Agents, Hormonal; Antineoplastic Agents; Contraceptives, Oral, Synthetic; Contraceptives, Oral