Sperm Phenotype and Differentially Methylated Regions

Infertility, Male Clinical Trial

— Epigenetics  

Official title:

Association Between Anogenital Distance, Sperm Phenotype and Epigenetics in Infertile Men.

Clinical Trial Details — Status: Completed

Administrative data

• NCT number: NCT05461079
• Other study ID #: RME12072017
• Status: Completed
• Start date: November 1, 2017
• Est. completion date: December 31, 2023

Study information

• Verified date: March 2024
• Source: University of Basel
• Contact: n/a
• Is FDA regulated: No
• Study type: Observational

Clinical Trial Summary

Testicular dysgenesis syndrome (TDS) is known to cause epigenetic abnormalities in spermatozoa. Anogenital distance (AGD) is considered to be a suitable clinical marker of TDS, but the direct link between AGD and epigenetic abnormalities is still missing.

Infertile men (n=10) presenting with shortened AGD and a control group of normal semen donors (n=10) with normal AGD will then be asked to provide one semen sample each. Using a flow cytometer and sorter (FACS) their spermatozoa will be sorted into populations of spermatozoa with/without DNA fragmentation or with/without chromatin decondensation. These sorted populations of spermatozoa will then be examined for differences in epigenetic imprinting differences using whole genome expression analysis. Whereas the sorting of spermatozoa will be carried out in Basel, the epigenetic analysis will be carried at the University of Geneva.

Description:

A subset of 10 men with shortened AGD (together with a control group of 10 fertile donors with normal AGD) will be asked to provide up to three semen samples, each of which then will be sorted with FACS into subpopulations with/without DNA fragmentation and into subpopulations with/without chromatin decondensation.

Spermatozoa with fragmented DNA will be separated through FACS-sorting of spermatozoa with intact DNA using the YoPro 1-dye, which has been shown to correlate significantly with the degree of DNA fragmentation in the nuclei of sperm.

In addition, spermatozoa with abnormal chromatin remodelling will be separated through sorting from spermatozoa with condensed chromatin using the fluorochrome chromomycin A3 (CMA3), which competes for protamin for binding to the minor groove of DNA thereby correlating with the persistence of histones in the sperm nuclei. Pilot experiments have demonstrated the highly significant and close correlation of CMA3 with anilin blue staining. Anilin blue staining is not suitable for the sorting experiment, because it requires fixation of the spermatozoa. Sorting based on CMA3 can be carried out with living spermatozoa.

The sorted and anonymized samples will then be sent frozen in dry ice to a laboratory at the University of Geneva for the assessment of differences in the epigenetic imprinting of the DNA using whole genome expression studies.

Recruitment information / eligibility

• Status: Completed
• Enrollment: 60
• Est. completion date: December 31, 2023
• Est. primary completion date: January 30, 2021
• Gender: Male
• Age group: 20 Years to 55 Years

Eligibility

Inclusion Criteria:

• infertile men with known anogenital distance

Exclusion Criteria:

• sperm concentration must be more than 15 million/ml to allow appropriate sorting with flow cytometry...

Related Conditions & MeSH terms

• Infertility
• Infertility, Male

Intervention

Diagnostic Test:
• obtention of up to three semen samples
sorting of spermatozoa with flow cytometry. In the presence of insufficient numbers of spermatozoa after sorting (<15 mill), up to three semen samples will be collected.

Locations

Country	Name	City	State
Switzerland	Christian De Geyter	Basel

Sponsors (2)

Lead Sponsor	Collaborator
University of Basel	University of Geneva, Switzerland

Country where clinical trial is conducted

Switzerland, 

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	anogenital distance and epigenetics	number of differentially methylated transposable regulatory sequences in the genome of sorted spermatozoa.	6 months
Secondary	sperm phenotype 1	based on conventional semen analysis: concentration of spermatozoa (in mill/ml).	6 months
Secondary	sperm phenotype 2	based on conventional semen analysis: progressive motility (in %).	6 months
Secondary	sperm phenotype 3	based on conventional semen analysis: normal morphology (in %), staining).	6 months
Secondary	sperm phenotype 4	chromatin decondensation (as given by % of CMA3 staining).	6 months
Secondary	sperm phenotype 5	DNA fragmentation (% of YoPro 1-staining).	6 months