Impact of Ionizing Treatment on the Nuclear Structure of Human Spermatozoa.

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.

Study Overview

• Status: Unknown
• Conditions: Sperm Preservation; Irradiated Semen
• Intervention / Treatment: Other: Cryopreserved semen; Other: Acute and low irradiation; Other: Long and low irradiation; Other: Long and medium irradiation

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.

• Study Type: Observational
• Enrollment (Anticipated): 200

Contacts and Locations

Study Locations

• France
  • Clermont-Ferrand, France, 63000
    • Recruiting
    • CHU de Clermont-Ferrand
    • Contact: Lise Laclautre, PharmD; Phone Number: 33 04 73 75 49 62; Email: promo_interne_drci@chu-clermontferrand.fr
    • Principal Investigator: Hanaé PONS-REJRAJI, PhD
    • Sub-Investigator: Florence BRUGNON, MD, PhD
    • Sub-Investigator: Cyril BOUCHE, PhD
    • Sub-Investigator: Aurélie VEGA, PhD
    • Sub-Investigator: Philippe VAGO, MD, PhD
    • Sub-Investigator: Andreï TCHIRKOV, MD, PhD
    • Sub-Investigator: Céline PEBREL-RICHARD, MD
    • Sub-Investigator: Gaëlle SALAUN, MD, PhD
    • Sub-Investigator: Lauren VERONESE, MD, PhD
    • Sub-Investigator: Laetitia GOUAS, MD, PhD
    • Sub-Investigator: Carole GOUMY, MD, PhD

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 45 years (ADULT)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: Male

• 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

Description

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:

-

Study Plan

How is the study designed?

Number of groups / cohorts

5

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
control arm "fresh"; Fresh semen treated without irradiation before cryopreservation	Other: Cryopreserved semen; sperm will be frozen in Cryosperm® cryoprotectant medium (Origio). The samples will be packaged in previously identified high security straws (Cryobiosystem). Slow freezing of the straws will be carried out using the Nanodigicool® programmable device (Cryobiosystem).
control arm "cryopreserved"; cryopreserved semen without irradiation (n=60)	Other: Cryopreserved semen; sperm will be frozen in Cryosperm® cryoprotectant medium (Origio). The samples will be packaged in previously identified high security straws (Cryobiosystem). Slow freezing of the straws will be carried out using the Nanodigicool® programmable device (Cryobiosystem).
acute irradiation at low dose; Pelvis scanning : acute irradiation (1 or 2 second) using low dose (n=30),	Other: Acute and low irradiation; sperm sample will be placed on the scanner's processing table to be exposed for 1 to 2 seconds. Several dose levels will then be made in order to limit the value of 17 mGy
long irradiation at low dose; Bone scintigraphy : long irradiation (3h) using low dose (n=30),	Other: Long and low irradiation; sperm sample will be exposed to gamma radiation by adding a solution of Tc99m for 3 hours.
long irradiation at medium dose; Radioactive Iodine 131 therapy (RAT) applied for thyroid cancer : long irradiation (3h) using medium dose (n=60),	Other: Long and medium irradiation; sperm sample will be exposed to gamma radiation by adding a solution of I131 for 3 hours

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
To measure in vitro the impact of an ionizing radiation treatment on sperm nuclear quality and in particular the size of telomeres.	To achieve this goal, we will expose human sperm to an irradiation dose of I131 to mimic the irradiation of gonads and sperm. Reliquat of semen will be cryopreserved (control condition). After thawing, the same sample will be subdivised into 2 groups in order to analyze the sperm quality : After an irradiation dose of I131; After a cryopreservation	01/01/2018-01/01/2022

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
To measure in vitro the impact of an irradiation dose of I131 on sperm nuclear quality, notably on STL.	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. Semen samples will be 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: 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.	01/01/2018-01/01/2022
to measure RAT impact on sperm parameters (vitality, motility and morphology)	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. Semen samples will be 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.; 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;	01/01/2018-01/01/2022
To measure in vitro the impact of cryopreservation on sperm nuclear quality, notably on STL, chromatin organisation and nucleus morphology	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. Semen samples will be 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: 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.	01/01/2018-01/01/2022
to evaluate RAT impact on human sperm cells in comparison with cryopreservation	We will compare effects induced by RAT and cryopreservation on each sperm sample. In order to limit the bias, for each irradiation treatment, a sample of sperm cells (cryopreserved control) will be treated under identical conditions.	01/01/2018-01/01/2022
to evaluate the impact of different doses and types of irradiation on human sperm cells	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. Semen samples will be 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. Irradiation exposure will be realised under 3 conditions: acute and low irradiation : sperm sample will be placed on the scanner's processing table to be exposed for 1 to 2 seconds. Several dose levels will then be made in order to limit the value of 17 mGy.; long and low irradiation: sperm sample will be exposed to gamma radiation by adding a solution of Tc99m for 3 hours.; long and medium irradiation : sperm sample will be exposed to gamma radiation by adding a solution of I131 for 3 hours	01/01/2018-01/01/2022
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.	We will correlate the effects induced by on vitality, motility and morphology and nuclear sperm parameters (DNA fragmentation and oxidation, chromatin condensation and organisation, nucleus morphology and STL.	01/01/2018-01/01/2022

Collaborators and Investigators

• Sponsor: University Hospital, Clermont-Ferrand
• Collaborators: Agence de La Biomédecine
• Investigators: Study Chair: Hanae PONS-REJRAJI, PhD, CHU de Clermont-Ferrand

Study record dates

Study Major Dates

• Study Start (ACTUAL): January 1, 2018
• Primary Completion (ANTICIPATED): January 1, 2022
• Study Completion (ANTICIPATED): January 1, 2022

Study Registration Dates

• First Submitted: January 15, 2021
• First Submitted That Met QC Criteria: January 15, 2021
• First Posted (ACTUAL): January 20, 2021

Study Record Updates

• Last Update Posted (ACTUAL): January 20, 2021
• Last Update Submitted That Met QC Criteria: January 15, 2021
• Last Verified: January 1, 2021

More Information

Terms related to this study

• Keywords: Telomere length; Thyroid Cancer; Fertility Preservation; Ionizing radiation; Human sperm; Nuclear alterations; DNA Fragmentation and Oxidation; Chromatin decondensation; Conventional sperm parameters
• Other Study ID Numbers: 2020 PONS-REJRAJI

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: UNDECIDED

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No