VATES: Relationship Between Sperm Head Vacuoles and Sperm DNA Alterations in Infertile Men

Sponsor

University Hospital, Rouen (Other)

Overall Status

Completed

CT.gov ID

NCT02006446

Collaborator

(none)

200

Enrollment

Locations

Actual Duration (Months)

100

Patients Per Site

2.7

Patients Per Site Per Month

Study Details

Study Description

Brief Summary

In men presenting sperm alterations, the selection of genetically undamaged spermatozoa need to be improved in order to increase the success of assisted reproduction treatments.

The aim of this study is to determine whether the presence of sperm head vacuoles is associated with sperm DNA alterations.

Condition or Disease	Intervention/Treatment	Phase
Male Infertility

Detailed Description

The use of intracytoplasmic sperm injection (ICSI) has greatly improved the treatment of severe male infertility, especially for men with oligo-astheno-teratozoospermia (OAT). This in vitro fertilization procedure allows the direct injection of a single spermatozoon into an oocyte. The sperm used for ICSI is selected under a microscope at a 400x magnification. Several studies have reported that de novo chromosomal abnormalities are increased in children born from ICSI , thereby raising the question of the genetic quality of spermatozoa used for ICSI.

A method called MSOME (high magnification Motile Sperm Organelle Morphology Examination), which allows the detailed morphological evaluation of motile spermatozoa in real time and under high magnification (6600x), was developed in 2001. With this technique, fine morphological abnormalities - mainly vacuoles in the head of spermatozoa - were detected. The use of MSOME for the detection of morphologically normal sperm for ICSI gave rise to a technique called IMSI (Intracytoplasmic Morphologically Selected sperm Injection). Higher pregnancy rates were obtained with IMSI compared to ICSI and sperm head vacuoles were found to negatively affect assisted reproduction success rates and embryo development. The use of IMSI also decreases the risk of sex chromosome aneuploidy in embryos .

Several authors found that the presence of large vacuoles in the sperm head correlates with several nuclear alterations: DNA fragmentation , abnormal chromatin condensation and aneuploidy . However, these studies are controversial and were performed on few spermatozoa. In order to improve the selection of spermatozoa with a normal chromosomal content, it is essential not only to confirm the existence of a relationship between sperm head vacuoles and altered sperm nuclear quality but also to better characterize these alterations.

The main goal of this study is to investigate the correlation between sperm head vacuole areas and sperm aneuploidy rates in men with isolated teratozoospermia or OAT. Vacuole areas will be measured by MSOME and aneuploidy rates by FISH for chromosomes 18, X and Y. Moreover, the correlation between sperm head vacuole areas and other nuclear alterations (DNA fragmentation, abnormal chromatin condensation, telomere abnormalities) will be evaluated. Semen samples from 200 patients recruited over a 2-year period will be collected, stored and analyzed.

This study involves the collection of relevant medical information. The computer website for the patient database is secure and protected by a password. The information will be entered and only be viewed by the investigators. On-site monitoring visits will be conducted throughout the study.

Study Design

Study Type:

Observational [Patient Registry]

Actual Enrollment :

200 participants

Observational Model:

Cohort

Time Perspective:

Prospective

Official Title:

Are Sperm Head Vacuoles Associated With Sperm Nuclear Alterations?

Actual Study Start Date :

Dec 1, 2013

Actual Primary Completion Date :

Dec 1, 2016

Actual Study Completion Date :

Jan 1, 2017

Arms and Interventions

Arm	Intervention/Treatment
VATES men with altered spermograms (isolated teratozoospermia or oligo-astheno-teratozoospermia)

Outcome Measures

Primary Outcome Measures

Correlation coefficient between mean vacuole areas in sperm heads (measured by MSOME) and sperm aneuploidy rates for chromosomes X, Y and 18 (evaluated by FISH) [up to 30 months]

Secondary Outcome Measures

Total sperm count [1 day]
Percentage of mobile spermatozoa [1 day]
Percentage of morphologically abnormal spermatozoa [1 day]
Mean vacuole area threshold (measured with Receiver Operating Characteristic curves) [up to 30 months]
Correlation coefficient between vacuole areas and sperm DNA fragmentation (evaluated by TUNEL analysis) [up to 30 months]
Correlation coefficient between vacuole areas and abnormal chromatin condensation (evaluated by aniline blue staining) [up to 30 months]
Correlation coefficient between vacuole areas and telomere number, distribution and length (evaluated by quantitative FISH) [up to 30 months]

Eligibility Criteria

Criteria

Ages Eligible for Study:

18 Years and Older

Sexes Eligible for Study:

Male

Accepts Healthy Volunteers:

Yes

Inclusion Criteria:

men
18 years old or more
affiliated to social security
with isolated teratozoospermia or oligo-astheno-teratozoospermia according to the WHO guidelines (2010):
sperm concentration < 15 million per ml or sperm number < 39 million per ejaculate
progressive motility < 35%
morphologically normal forms < 50% according to the modified David's classification (Auger et al., 2001
mobile spermatozoa selected by density gradient centrifugation > 1 million, in previous spermograms
with normal constitutional karyotypes (46, XY)
signed an informed consent

Exclusion Criteria:

with abnormal constitutional karyotypes
with a major alteration of sperm parameters, with mobile spermatozoa selected by density gradient centrifugation < 1 million
under tutorship or guardianship by court order

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	Laboratoire de spermiologie, CHRU de Lille	Hôpital Calmette CHRU de Lille		France
2	Laboratoire de Biologie de la Reproduction - CECOS - EA 4308, CHU - Hôpitaux de Rouen	Rouen		France	76031