Tachykinin and Kisspeptin Expression in Human Granulosa and Cumulus Cells

Development of a New Diagnostic Tool to Assess Oocyte Quality Based on Tachykinin and Kisspeptin Expression Analysis in Human Granulosa and Cumulus Cells

The purpose of this project is to study the presence of expression differences - at RNA and protein level - of different members of the tachykinin family and kisspeptin and their receptors, between fertile women and infertile patients with different etiologies.

Study Overview

• Status: Completed
• Conditions: Infertility
• Intervention / Treatment: Procedure: Ovarian puncture

• Study Type: Interventional
• Enrollment (Actual): 236
• Phase: Not Applicable

Contacts and Locations

Study Locations

• Spain
  • Sevilla, Spain, 41011
    • IVI Sevilla
  • Sevilla, Spain, 41011
    • Instituto de Investigaciones Químicas

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 45 years (ADULT)
• Accepts Healthy Volunteers: Yes
• Genders Eligible for Study: Female

Description

Two groups of subjects (each one with different criteria):

1. Oocyte donors

   Inclusion Criteria:

   • Age between 18 and 34 years
   • Normal caryotype
   • Normal psychological test

   Exclusion Criteria:

   • Presence of hereditary diseases
   • Beta-thalassemia
   • Cystic fibrosis
   • Human immunodeficiency virus (HIV), Hepatitis B virus (HBV), Hepatitis C virus (HCV)
2. Infertility patients

Inclusion Criteria:

• Age between 18 and 45 years
• One of the following etiologies:
• Patient with PCOS according to Rotterdam Criteria or
• Patients with endometriosis stage I-IV or
• Patients with low ovarian response according to Bologna criteria or
• Patients with advances maternal age (between 38 and 45 years)

Study Plan

How is the study designed?

Design Details

• Primary Purpose: BASIC_SCIENCE
• Allocation: NON_RANDOMIZED
• Interventional Model: SINGLE_GROUP
• Masking: NONE

Number of Arms

5

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
ACTIVE_COMPARATOR: Donors; Oocyte donors without infertility problems	Procedure: Ovarian puncture; Donors and patients are subjected to ovarian puncture as part of their donation procedure or IVF treatment. Oocyte are retrieved in this procedure along with granulosa and cumulus cells. Donors and patients sign an informed consent in order to donate their granulosa and cumulus cells for the study.
EXPERIMENTAL: PCOS; Patients with Polycystic Ovary Syndrome (PCOS) according to Rotterdam criteria	Procedure: Ovarian puncture; Donors and patients are subjected to ovarian puncture as part of their donation procedure or IVF treatment. Oocyte are retrieved in this procedure along with granulosa and cumulus cells. Donors and patients sign an informed consent in order to donate their granulosa and cumulus cells for the study.
EXPERIMENTAL: Endometriosis; Patients with endometriosis (I-IV)	Procedure: Ovarian puncture; Donors and patients are subjected to ovarian puncture as part of their donation procedure or IVF treatment. Oocyte are retrieved in this procedure along with granulosa and cumulus cells. Donors and patients sign an informed consent in order to donate their granulosa and cumulus cells for the study.
EXPERIMENTAL: Age; Patients with advanced maternal age (38 years or more)	Procedure: Ovarian puncture; Donors and patients are subjected to ovarian puncture as part of their donation procedure or IVF treatment. Oocyte are retrieved in this procedure along with granulosa and cumulus cells. Donors and patients sign an informed consent in order to donate their granulosa and cumulus cells for the study.
EXPERIMENTAL: Low ovarian response; Patients with low ovarian response according to Bologna criteria	Procedure: Ovarian puncture; Donors and patients are subjected to ovarian puncture as part of their donation procedure or IVF treatment. Oocyte are retrieved in this procedure along with granulosa and cumulus cells. Donors and patients sign an informed consent in order to donate their granulosa and cumulus cells for the study.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Expression level of Substance P (SP)	Expression analysis at mRNA (mRNA = messenger ribonucleic acid) and protein level using techniques of real time quantitative PCR (PCR = polymerase chain reaction), immunocytochemistry and western blot	4 years
Expression level of neurokinin A (NKA)	Expression analysis at mRNA and protein level using techniques of real time quantitative PCR, immunocytochemistry and western blot	4 years
Expression level of neurokinin B (NKB)	Expression analysis at mRNA and protein level using techniques of real time quantitative PCR, immunocytochemistry and western blot	4 years
Expression level of hemokinin 1 (HK-1)	Expression analysis at mRNA and protein level using techniques of real time quantitative PCR, immunocytochemistry and western blot	4 years
Expression level of neurokinin 1 receptor (NK1R)	Expression analysis at mRNA and protein level using techniques of real time quantitative PCR, immunocytochemistry and western blot	4 years
Expression of neurokinin 2 receptor (NK2R)	Expression analysis at mRNA and protein level using techniques of real time quantitative PCR, immunocytochemistry and western blot	4 years
Expression level of neurokinin 3 receptor (NK3R)	Expression analysis at mRNA and protein level using techniques of real time quantitative PCR, immunocytochemistry and western blot	4 years
Expression level of kisspeptin 1 (KISS1)	Expression analysis at mRNA and protein level using techniques of real time quantitative PCR, immunocytochemistry and western blot	4 years
Expression level of kisspeptin 1 receptor (KISS1R)	Expression analysis at mRNA and protein level using techniques of real time quantitative PCR, immunocytochemistry and western blot	4 years

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Patient or donor age	Years	4 years
Patient or donor body mass index	kg/m2	4 years
Years of infertility	Number of years that the patient has been trying to have a child without success	4 years
Estradiol level (day of ovulation induction)	Estradiol level in blood (pg/mL), measured the day in which ovulation is induced	4 years
Progesterone level (day of ovulation induction)	Progesterone level in blood (ng/mL), measured the day in which ovulation is induced	4 years
Number of oocytes retrieved in the ovarian puncture		4 years
Number of mature oocytes (metaphase II) obtained in the ovarian puncture		4 years
Fertilization rate	Proportion of oocytes correctly fertilized (2 pronuclei and 2 polar bodies) after the ICSI (ICSI = intracytoplasmic sperm injection) procedure	4 years
Embryo quality	Embryo quality at day 3 of embryo development (of each embryo of the cohort). Quality assessed according to ASEBIR (ASEBIR = "Asociación Española para el estudio de la Biología de la Reproducción" = Spanish society for reproductive biology) criteria. Embryo quality grade A to D.	4 years
Daily and total dose of FSH (FSH = follicle stimulating hormone)	Daily dose of FSH for controlled ovarian stimulation. International Units (IU)	4 years
Daily and total dose of HMG (HMG = human menopausal gonadotropin)	Daily dose of HMG for controlled ovarian stimulation. International Units (IU)	4 years
Type of ovulation induction	Ovulation induction using 6500 IU of hCG (hCG = human chorionic gonadotropin) or 0.2 mg of triptorelin (Decapeptyl)	4 years

Collaborators and Investigators

• Sponsor: IVI Sevilla

Publications and helpful links

General Publications

• Garcia-Ortega J, Pinto FM, Fernandez-Sanchez M, Prados N, Cejudo-Roman A, Almeida TA, Hernandez M, Romero M, Tena-Sempere M, Candenas L. Expression of neurokinin B/NK3 receptor and kisspeptin/KISS1 receptor in human granulosa cells. Hum Reprod. 2014 Dec;29(12):2736-46. doi: 10.1093/humrep/deu247. Epub 2014 Oct 14.
• Garcia-Ortega J, Pinto FM, Prados N, Bello AR, Almeida TA, Fernandez-Sanchez M, Candenas L. Expression of Tachykinins and Tachykinin Receptors and Interaction with Kisspeptin in Human Granulosa and Cumulus Cells. Biol Reprod. 2016 Jun;94(6):124. doi: 10.1095/biolreprod.116.139881. Epub 2016 May 4.

Study record dates

Study Major Dates

• Study Start: January 1, 2014
• Primary Completion (ACTUAL): July 1, 2019
• Study Completion (ACTUAL): July 1, 2019

Study Registration Dates

• First Submitted: August 12, 2016
• First Submitted That Met QC Criteria: August 19, 2016
• First Posted (ESTIMATE): August 25, 2016

Study Record Updates

• Last Update Posted (ACTUAL): February 13, 2020
• Last Update Submitted That Met QC Criteria: February 11, 2020
• Last Verified: February 1, 2020

More Information

Terms related to this study

• Keywords: Infertility; Kisspeptin; Tachykinin
• Additional Relevant MeSH Terms: Infertility
• Other Study ID Numbers: IVISEVILLA