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NCT Number: NCT05461079

Sperm Phenotype and Differentially Methylated Regions

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.

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Key information

Conditions

Infertility, Male Genital Diseases Genital Diseases, Male Infertility Male Urogenital Diseases Urogenital Diseases

Age range

20 year–55 year

Sex eligibility

Male

Study type

Observational

Primary location

Christian De Geyter

Basel, 4031, Switzerland

About this study

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.

Who can participate

Only the study team can determine whether someone qualifies for participation.

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...

Treatment and study plan

obtention of up to three semen samples

Diagnostic Test

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.

Other names: conventional semen analysis followed by sorting

Primary outcomes

  1. anogenital distance and epigenetics

    Time frame: 6 months

    number of differentially methylated transposable regulatory sequences in the genome of sorted spermatozoa.

Secondary outcomes

  1. sperm phenotype 1

    Time frame: 6 months

    based on conventional semen analysis: concentration of spermatozoa (in mill/ml).

  2. sperm phenotype 2

    Time frame: 6 months

    based on conventional semen analysis: progressive motility (in %).

  3. sperm phenotype 3

    Time frame: 6 months

    based on conventional semen analysis: normal morphology (in %), staining).

  4. sperm phenotype 4

    Time frame: 6 months

    chromatin decondensation (as given by % of CMA3 staining).

  5. sperm phenotype 5

    Time frame: 6 months

    DNA fragmentation (% of YoPro 1-staining).

Sponsors and collaborators

Lead sponsor

University of Basel

Other

Collaborators

  • University of Geneva, Switzerland

Registry information

Official study title

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

Acronym: Epigenetics

Important dates

Study start
2017
Primary completion
2021
Study completion
2023
First posted
Jul 15, 2022
Registry last updated
Mar 20, 2024

OpenTrials presents study information sourced from ClinicalTrials.gov. The official registry record should be consulted for the latest information.

View the official ClinicalTrials.gov record (opens in a new tab)

This listing is for discovery and informational purposes only. It is not medical advice, does not guarantee that a study is recruiting, and does not determine eligibility. Contact the study team and a qualified healthcare professional when considering participation.

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