Clinical trial NCT05958914: Neuropeptide Expression During the Ovarian Cycle and in Patients With PCOS
Official title: Evaluation of Neuropeptide Expression During the Ovarian Cycle and in Patients With PCOS: Pilot Study
Study type: INTERVENTIONAL
Overall status: RECRUITING
Phase: NA
Conditions: PCOS
Keywords: PCOS
neuropeptides
enkephalins
SP
CGRP
VIP
Lead sponsor: University Of Perugia
Sponsor class: OTHER
First posted: 2023-07-25
Enrollment: 45
Sex: FEMALE
Minimum age (years): 18
Maximum age (years): 45
Standard ages: ADULT
Accepts healthy volunteers: yes
Allocation: NON_RANDOMIZED
Intervention model: PARALLEL
Intervention model description: The study involves an enrollment of 45 total patients, referred to the Medically assisted reproduction Center, divided into 3 groups, of 15 patients each: Group 1 (control) women with regular ovarian cycle; Group 2- non-PCOS women undergoing ovarian stimulation and ICSI treatment; Group 3- women with PCOS undergoing ovarian stimulation and ICSI treatment
Primary purpose: BASIC_SCIENCE

Brief summary: The goal of this clinical trial is to characterize PCOS in terms of neuropeptide expression and to make the comparison between women with and without PCOS by going to analyze venous blood samples taken during the three phases of the ovarian cycle and follicular fluid, taken during the ovulatory phase as part of the oocyte pick-up. Specifically, the expression of neuropeptides such as CGRP, SP, VIP and enkephalins in the three phases of the ovarian cycle (follicular phase, ovulatory phase and luteal phase) will be evluated in three groups of women afferent to the medically assisted reproduction centre. In particular, Group 1 (control) will include women with regular ovarian cycle; Group 2 will include non-PCOS women undergoing ovarian stimulation and ICSI treatment; finally Group 3 will include women with PCOS undergoing ovarian stimulation and ICSI treatment.

The main question\[s\] it aims to answer are:

* Is there a fluctuation in blood neuropeptides concentrations of women with normal ovarian cycles during the three phases of the ovarian cycle?
* Do PCOS women has altered levels of blood and follicolar fluid concentration of neuropepides with respect to non-PCOS individuals?
* Is blood and follicolar fluid concentration of neuropeptide modulated by protocols of ovarian stimulation?

Participants of Group 1 will follow the ovarian monitoring protocol, during which blood samples will be taken at the three phases of the ovarian cycle.

Participants of Group 2 and 3 will undergo ovarian stimulation and ICSI treatment, followed by blood and follicular fluid sampling at the specified cycle phases.

Researchers will compare the control Group 1 with Group 2 and 3 to see if there is a significative difference in the concentration of blood neuropeptides between the three groups at the same phase of the ovarian cycle.

Moreover, they will evaluate if there is there significant different concentration of neuropeptides in follicolar fluid between group 2 and 3 during the ovulatory phase.

Detailed description: This project aims to investigate the neurosensory mechanisms underlying one of the most common disorders related to folliculogenesis, such as PCOS. The study aims to characterize PCOS in terms of neuropeptide expression and to make the comparison between women with and without PCOS, by going to analyze venous blood samples taken during the three phases of the ovarian cycle and follicular fluid, taken during the ovulatory phase as part of the oocyte pick-up. Specifically, the expression of neuropeptides such as CGRP, SP, VIP and enkephalins in the three phases of the ovarian cycle (follicular phase, ovulatory phase and luteal phase) in the three groups of women considered will be evaluated.

Study design:

The study will enroll 45 total patients, referred to the PMA Center, divided into 3 groups of 15 patients each: Group 1 (control) women with regular ovarian cycle; Group 2- non-PCOS women undergoing ovarian stimulation and ICSI treatment; Group 3- women with PCOS undergoing ovarian stimulation and ICSI treatment.

The study will have a total duration of 24 months divided as follows:

* Pre-phase monitoring and enrollment of subjects (20 months).

  * Group 1 will follow the ovarian monitoring protocol, during which blood samples will be taken at the three stages of the ovarian cycle.
  * Group 2 and 3 will undergo ovarian stimulation and ICSI treatment, taken with blood and follicular fluid samples at the specified cycle phases.
* Performance of chemical/biochemical analysis performed at the electrophysiological laboratory of the University of Perugia (2 months).
* Performance of statistical tests and processing of the data obtained (2 months)

Subjects will be given a 4 mL venous blood draw as per regular clinical practice to perform hormone dosing during ovarian cycle monitoring.

No diagnostic tests outside of routine clinical practice are planned, and patients will not be required to travel to the PMA Center for additional visits beyond those required by regular clinical practice.

Subjects and Recruitment Location: 45 patients will be recruited: 15 women with a diagnosis of PCOS and 30 non-PCOS women at the medically assisted reproduction Center of the Perugia Hospital (Head: Prof. S. Gerli).

MATERIALS AND METHODS:

GROUP 1: Patients, as per normal clinical practice, will undergo three blood draws in the follicular, ovulatory and luteal phases.

GROUP 2 and 3: Patients will undergo ovarian stimulation treatment with gonadotropins and egg retrieval. As part of this process, as per clinical practice, blood and follicular fluid sampling will be performed.

Methods Biological Material Collection: During ovarian cycle monitoring, a venous blood sampling will be performed in the morning after at least 10 hours of fasting in patients during the phases described in Table 1. A follicular fluid sample will also be taken during oocyte pick-up (group 2 and 3). Specifically, an aliquot of blood sample, which follicular fluid will be evaluated to assay the level of neuropeptides (CGRP, SP, VIP and enkephalins). The level of CGRP will be measured using an extraction-free enzyme immunoassay kit (Peninsula laboratories LLC, CA, USA) following the manufacturer's protocol. For the quantitative measurement of human substance P, the Human Substance P Enzyme ELISA Assay Kit (ELK1453-96T - Twin Elix srl) will be used. For VIP, the ELISA Kit for Human VIP (ELK1453-96T - Twin Elix srl) will be used, and for the enkephalins assay, the ELISA Kit for Human Enkephalins (ELK5246-96T - Twin Elix srl) will be used. Neuropeptide assays will be conducted at the Physiology Laboratory of the Department of Chemistry, Biology and Biotechnology, University of Perugia, in collaboration with Prof. Fioretti.

Sera will also be analyzed for fasting glucose, triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL), insulin, dehydroepiandrosterone sulfate (DHEAS), sex hormone-binding globulin (SHBG), progesterone, estradiol, follicle-stimulating hormone (FSH), LH and total testosterone. Serum concentrations of glucose (hexokinase), total cholesterol (CHOD-PAP) and triglycerides (GPO-PAP) will also be measured using standard enzymatic methods (Roche Diagnostics, IN, USA) with a fully automated analyzer (Roche Modular PE, Roche Diagnostics, IN, USA).

Eligibility criteria: Inclusion Criteria:

* infertile women
* BMI between 18 and 30 kg/m2,
* basal FSH \< 10 IU/L,
* number of antral follicles (2-10 m) per ovary \> 10,
* regular uterine cavity assessed by hysterosalpinography, sonohysterography or hysteroscopy, and hematologic and biochemical parameters within normal limits will be included in the study, euthyroid status (with or without treatment)
* In case of women with PCOS, it should be diagnosed according to the Rotterdam criteria (The Rotterdam ESHRE/ASRM-sponsored PCOS consensus workshop group (2004) Revised 2003 consensus on diagnostic criteria and longterm health risks related to polycystic ovary syndrome (PCOS).
* No restrictions regarding the indication of infertility.

Exclusion Criteria:

* presence of ovaries inaccessible to oocyte pick-up,
* presence of sactosalpinx, heterologous fertilization,
* contraindication to pregnancy,
* atypical genital discharge of unspecified cause,
* uncontrolled dystothyroidism, presence of neoplasms,
* severe alteration of liver or kidney function, taking medications that may interfere with the study.

Primary outcomes:
measure: Evaluation of the normal fluctuation of CGRP during the ovarian cycle in non-PCOS women
timeFrame: 5 days postmenstrual, 14 days postmenstrual, 18 days postmenstrual
measure: Evaluation of the normal fluctuation of SP during the ovarian cycle in non-PCOS women
timeFrame: 5 days postmenstrual, 14 days postmenstrual, 18 days postmenstrual
measure: Evaluation of the normal fluctuation of VIP during the ovarian cycle in non-PCOS women
timeFrame: 5 days postmenstrual, 14 days postmenstrual, 18 days postmenstrual
measure: Evaluation of the normal fluctuation of enkephalins during the ovarian cycle of non PCOS women
timeFrame: 5 days postmenstrual, 14 days postmenstrual, 18 days postmenstrual
Secondary outcomes:
measure: Evaluation of the fluctuation of CGRP during the ovarian cycle in non-PCOS women undergoing ICSI treatment
timeFrame: 5 days before egg pick-up, day of egg pick up, 10 days after egg pick-up
measure: Evaluation of the fluctuation of SP during the ovarian cycle in non-PCOS women undergoing ICSI treatment
timeFrame: 5 days before egg pick-up, day of egg pick up, 10 days after egg pick-up
measure: Evaluation of the fluctuation of enkephalins during the ovarian cycle in non-PCOS women undergoing ICSI treatment
timeFrame: 5 days before egg pick-up, day of egg pick up, 10 days after egg pick-up
measure: Evaluation of the fluctuation of VIP during the ovarian cycle in non-PCOS women undergoing ICSI treatment
timeFrame: 5 days before egg pick-up, day of egg pick up, 10 days after egg pick-up
measure: Evaluation of the fluctuation of CGRP during the ovarian cycle in PCOS women undergoing ICSI treatment
timeFrame: 5 days before egg pick-up, day of egg pick up, 10 days after egg pick-up
measure: Evaluation of the fluctuation of SP during the ovarian cycle in PCOS women undergoing ICSI treatment
timeFrame: 5 days before egg pick-up, day of egg pick up, 10 days after egg pick-up
measure: Evaluation of the fluctuation of enkephalins during the ovarian cycle in PCOS women undergoing ICSI treatment
timeFrame: 5 days before egg pick-up, day of egg pick up, 10 days after egg pick-up
measure: Evaluation of the fluctuation of VIP during the ovarian cycle in PCOS women undergoing ICSI treatment
timeFrame: 5 days before egg pick-up, day of egg pick up, 10 days after egg pick-up
measure: Comparison of CGRP blood concentration between PCOS women and non-PCOS women undergoing ICSI treatment
timeFrame: 5 days before egg pick-up, day of egg pick up, 10 days after egg pick-up
measure: Comparison of SP blood concentration between PCOS women and non-PCOS women undergoing ICSI treatment
timeFrame: 5 days before egg pick-up, day of egg pick up, 10 days after egg pick-up
measure: Comparison of enkephalins blood concentration between PCOS women and non-PCOS women undergoing ICSI treatment
timeFrame: 5 days before egg pick-up, day of egg pick up, 10 days after egg pick-up
measure: Comparison of VIP blood concentration between PCOS women and non-PCOS women undergoing ICSI treatment
timeFrame: 5 days before egg pick-up, day of egg pick up, 10 days after egg pick-up
measure: Comparison of CGRP blood concentration between non-PCOS women with normal ovarian cycle and non-PCOS women undergoing ICSI treatment
timeFrame: 7 days after menstruation, 14 days after menstruation, 20 days after menstruation
measure: Comparison of SP blood concentration between non-PCOS women with normal ovarian cycle and non-PCOS women undergoing ICSI treatment
timeFrame: 7 days after menstruation, 14 days after menstruation, 20 days after menstruation
measure: Comparison of enkephalins blood concentration between non-PCOS women with normal ovarian cycle and non-PCOS women undergoing ICSI treatment
timeFrame: 7 days after menstruation, 14 days after menstruation, 20 days after menstruation
measure: Comparison of VIP blood concentration between non-PCOS women with normal ovarian cycle and non-PCOS women undergoing ICSI treatment
timeFrame: 7 days after menstruation, 14 days after menstruation, 20 days after menstruation
measure: Comparison of CGRP concentration in follicolar fluid between non-PCOS women and PCOS women both undergoing ICSI treatment
timeFrame: 5 days before egg pick-up, day of egg pick up, 10 days after egg pick-up
measure: Comparison of SP concentration in follicolar fluid between non-PCOS women and PCOS women both undergoing ICSI treatment
timeFrame: 5 days before egg pick-up, day of egg pick up, 10 days after egg pick-up
measure: Comparison of enkephalins concentration in follicolar fluid between non-PCOS women and PCOS women both undergoing ICSI treatment
timeFrame: 5 days before egg pick-up, day of egg pick up, 10 days after egg pick-up
measure: Comparison of VIP concentration in follicolar fluid between non-PCOS women and PCOS women both undergoing ICSI treatment
timeFrame: 5 days before egg pick-up, day of egg pick up, 10 days after egg pick-up
Other outcomes:
Locations:
city: Perugia
country: Italy
facility: University of Perugia, Piazza Menghini 1, S. Andrea delle Fratte.
state: Umbria
Study officials:
affiliation: University Of Perugia
name: Sandro Gerli, Professor
role: PRINCIPAL_INVESTIGATOR
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