Clinical trial NCT04715828: Impact of Ionizing Treatment on the Nuclear Structure of Human Spermatozoa.
Official title: Impact of Ionizing Treatment on the Nuclear Structure of Human Spermatozoa.
Study type: OBSERVATIONAL
Overall status: UNKNOWN
Conditions: Sperm Preservation
Irradiated Semen
Keywords: Fertility Preservation
Ionizing radiation
Human sperm
Thyroid Cancer
Nuclear alterations
Telomere length
DNA Fragmentation and Oxidation
Chromatin decondensation
Conventional sperm parameters
Lead sponsor: University Hospital, Clermont-Ferrand
Sponsor class: OTHER
Collaborators: name: Agence de La Biomédecine
First posted: 2021-01-20
Enrollment: 200
Sex: MALE
Minimum age (years): 18
Maximum age (years): 45
Standard ages: ADULT
Accepts healthy volunteers: no
Observational model: COHORT
Time perspective: PROSPECTIVE
Sampling method: NON_PROBABILITY_SAMPLE
Study population: All measurements are made on surplus samples from men undergoing routine semen analysis at the Center for Reproductive Medicine, after receiving their written informed consent

Brief summary: Differentiated thyroid cancer is the third cause of cancer in young men of childbearing age. Its treatment by irradiation with Radioactive Iodine 131 therapy (RAT) could alter spermatogenesis and result in azoospermia and permanent infertility. A preventive gametes cryopreservation was recommended before RAT, but without mentioning a period of teratogenic risk transmissible to the offspring. To date, RAT impact on human sperm nucleus is poorly known or even unknown, notably on telomere length.

Our objective is to define RAT effects on human sperm nucleus by in vitro irradiation exposure of human spermatozoa to mimicking that of the gonads in the context of irradiation with iodine131 used for thyroid cancer. We will analyze standard sperm parameters, major DNA alterations and telomere length using molecular and cellular assays. Nucleus morphology and chromatin organization will also be analyzed using 3D bio-imaging. This study will permit to optimize the indications for the preservation of fertility.

Detailed description: Our main objective is to measure the in vitro impact of irradiation treatment on sperm nuclear quality such as DNA fragmentation and oxidation, chromatin condensation and organisation, nucleus morphology and notably sperm telomere length (STL).

The secondary objectives are :

* to measure RAT impact on sperm parameters (vitality, motility and morphology)
* to measure the impact of cryopreservation on chromatin organisation and nucleus morphology
* to evaluate RAT impact on human sperm cells in comparison with cryopreservation
* to evaluate the impact of different doses and types of irradiation on human sperm cells
* to establish relations between potential alterations of standards sperm parameters (vitality, motility and morphology) and nuclear sperm parameters (DNA fragmentation and oxidation, chromatin condensation and organisation, nucleus morphology and STL.

Our final goal is to provide a significant improvement of men fertility diagnosis and optimize our fertility preservation practices.

To this end, we will expose ejaculated human spermatozoa (n = 90) at different (low and moderate) doses of gamma or X photons to mimic gonads irradiation. Absorbed doses by the samples were calculated using the GATE Monte Carlo platform (version 8.2). Experiment geometry settings were modelled as three dimension voxelized volumes inside the software by assigning a shape, size, distance and density for all the volumes created. All measurements will be made on surplus samples from men undergoing routine semen analysis at the Center for Reproductive Medicine, after receiving their written informed consent. Semen samples are collected and subdivided into 3 arms to analyse sperm quality after:

* irradiation exposure (3 conditions);
* a freezing- thawing cycle;
* fresh state: negative control without treatment.

Before (fresh state) and after each treatment we will analyse:

* standard semen parameters (vitality, motility and morphology) in accordance with WHO, 2010;
* STL using Flow FISH, q-PCR and q-FISH;
* chromatin condensation (chromomycin A3);
* DNA oxidization (8-OHdG residues);
* DNA fragmentation (TUNEL);
* 3D nucleus structure using 3D bio-imaging.

Eligibility criteria: Inclusion Criteria:

* Man less than 45 years
* Man undergoing routine semen analysis at the Center for Reproductive Medicine, accepting the research protocol and signing the associated consent will be included in the protocol without distinction of physical criteria.

Exclusion Criteria:

-

Primary outcomes:
measure: To measure in vitro the impact of an ionizing radiation treatment on sperm nuclear quality and in particular the size of telomeres.
timeFrame: 01/01/2018-01/01/2022
Secondary outcomes:
measure: To measure in vitro the impact of an irradiation dose of I131 on sperm nuclear quality, notably on STL.
timeFrame: 01/01/2018-01/01/2022
measure: to measure RAT impact on sperm parameters (vitality, motility and morphology)
timeFrame: 01/01/2018-01/01/2022
measure: To measure in vitro the impact of cryopreservation on sperm nuclear quality, notably on STL, chromatin organisation and nucleus morphology
timeFrame: 01/01/2018-01/01/2022
measure: to evaluate RAT impact on human sperm cells in comparison with cryopreservation
timeFrame: 01/01/2018-01/01/2022
measure: to evaluate the impact of different doses and types of irradiation on human sperm cells
timeFrame: 01/01/2018-01/01/2022
measure: to establish relations between potential alterations of standards sperm parameters (vitality, motility and morphology) and nuclear sperm parameters (DNA fragmentation and oxidation, chromatin condensation and organisation, nucleus morphology and STL.
timeFrame: 01/01/2018-01/01/2022
Other outcomes:
Locations:
city: Clermont-Ferrand
country: France
facility: CHU de Clermont-Ferrand
Study officials:
affiliation: CHU de Clermont-Ferrand
name: Hanae PONS-REJRAJI, PhD
role: STUDY_CHAIR
References: