Sperm Phenotype and Differentially Methylated Regions

NCT ID: NCT05461079

Last Updated: 2024-03-20

Study Results

Results pending

The study team has not published outcome measurements, participant flow, or safety data for this trial yet. Check back later for updates.

Basic Information

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Recruitment Status

COMPLETED

Total Enrollment

60 participants

Study Classification

OBSERVATIONAL

Study Start Date

2017-11-01

Study Completion Date

2023-12-31

Brief Summary

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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.

Detailed Description

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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.

Conditions

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Infertility, Male

Study Design

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Observational Model Type

CASE_CONTROL

Study Time Perspective

PROSPECTIVE

Study Groups

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subfertile men

10 infertile men with shortened AGD (\< 40 mm)

obtention of up to three semen samples

Intervention Type 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.

fertile semen donors

10 fertile semen donors with normal AGD (\>40 mm)

obtention of up to three semen samples

Intervention Type 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.

Interventions

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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.

Intervention Type DIAGNOSTIC_TEST

Other Intervention Names

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conventional semen analysis followed by sorting

Eligibility Criteria

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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...
Minimum Eligible Age

20 Years

Maximum Eligible Age

55 Years

Eligible Sex

MALE

Accepts Healthy Volunteers

No

Sponsors

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University of Geneva, Switzerland

OTHER

Sponsor Role collaborator

University of Basel

OTHER

Sponsor Role lead

Responsible Party

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Christian De Geyter

Prof. Dr.med.

Responsibility Role PRINCIPAL_INVESTIGATOR

Locations

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Christian De Geyter

Basel, , Switzerland

Site Status

Countries

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Switzerland

Other Identifiers

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RME12072017

Identifier Type: -

Identifier Source: org_study_id

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