A Multimodal Study of the Relationship Between Exposure to Endocrine Disruptors and Occurrence of Hypospadias - HYPOLLUT (HYPOLLUT)

Exposure to Endocrine Disruptors and Occurrence of Hypospadias: Toxicological, Environmental and Hormonal Imbalance Study

This study aims to demonstrate the impact of Endocrine-Disrupting Chemicals (EDCs) on the risk of hypospadias incidence. It is a multicenter comparative case-control study, involving two groups. The first group consists of biological mothers who have given birth to children with hypospadias (Case Group), while the second group consists of biological mothers who have given birth to children without any malformations (Control Group). Through an integrative approach that combines a direct toxicological study of numerous pollutants present during pregnancy, and a comprehensive exposome assessment using validated tools, this study can significantly enhance our understanding and prevention of this malformation.

Study Overview

• Status: Recruiting
• Conditions: Hypospadias
• Intervention / Treatment: Other: Consultation visit (Visit 1); Other: Consultation visit (Visit 1)

Detailed Description

Background. Hypospadias is a birth defect of the external genital organs in boys and it ranks the second most common genital malformation in male newborns, following cryptorchidism. Its prevalence is increasing in certain global regions, with an estimated rate of 3.8 cases per 1000 male births. To date, the exact cause of hypospadias remains unknown, however genetic, hormonal, and environmental factors are likely involved. Medical and surgical treatment may be necessary. Furthermore, hypospadias is correlated with fertility issues and is also linked to testicular cancer.

Aim. After ruling out hypospadias with a genetic cause, the aim of this study is to evaluate any significant differences to environmental endocrine disrupting-chemicals (EDCs) exposure between biological mothers of children with hypospadias and those with children without malformation. It aims to demonstrate that this exposure (professional, occupational, environmental) leads to hormonal changes during the neonatal mini-puberty period.

Methods. This research will be conducted as a multicenter case-control study: mother and son with isolated anterior or middle hypospadias (Case Group) and mother and son without hypospadias (Control Group). The clinical investigator plans to enroll 200 patients.

A single visit will be performed. This consultation is part of the usual follow-up for children in the Case Group, while it is specific to the project for children in the Control Group.

During this visit, the investigator:

• will establish the diagnosis of hypospadias (for cases) or absence of genital anomaly (for controls)
• will lead an interview using a questionnaire and a job-exposure matrix to assess EDCs during pregnancy
• will take a hair sample from the mother to measure the substances accumulated during pregnancy
• and finally, will take a blood sample from the child for hormonal evaluation of mini-puberty, and another blood sample from child in the Case Group for analysis and the participation in a DNA collection

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

Contacts and Locations

• Study Contact: Name: Nicolas KALFA, Prof; Phone Number: 0467338784; Email: n-kalfa@chu-montpellier.fr

Study Locations

• France
  • Hérault
    • Montpellier, Hérault, France, 34090
      • Recruiting
      • CRMR DEVGEN CHU Lapeyronnie

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Child; Adult; Older Adult
• Accepts Healthy Volunteers: Yes

• Sampling Method: Non-Probability Sample

Study Population

Case Group: Biological mother and child with hypospadias Control group: Biological mother and child without hypospadias

Description

Non-specific Inclusion Criteria :

• Parents of legal age having signed a free and informed consent for the participation of their child
• Biological mother of a boy aged between 1 and 6 months
• Biological mother with a minimum hair length of 18 cm
• Biological mother who has signed a free and informed consent for her participation
• Biological mother and child affiliated with or beneficiaries of a national health insurance plan
• Biological mother who is fluent in written and spoken French

Specific Inclusion Criteria for Case Group:

- The child has an isolated anterior or middle hypospadias, without any other complex variations of genital development (borderline penile size, unilateral or bilateral cryptorchidism, retractile testes), without malformation syndrome and without identified genetic etiology

Specific Inclusion Criteria for Control Group:

- The child must not present any complex variations in genital development (hypospadias, borderline penis size, unilateral or bilateral cryptorchidism, retractile testes)

Exclusion Criteria:

• Child with another congenital anomaly or malformative syndrome
• Child with an endocrine pathology
• Biological mother or child under legal protection, guardianship, or curatorship
• Biological mother or child in the exclusion period of a previous study
• Biological mother or child included in another clinical study involving a drug

Specific Exclusion Criteria for Case Group:

Biological mother/child pairs if a genetic variant explaining hypospadias is found during genetic analysis.

Study Plan

How is the study designed?

Number of groups / cohorts

2

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
Case Group; Biological mother and son with hypospadias	Other: Consultation visit (Visit 1); During visit 1, a pediatric urologist or pediatric endocrinologist will perform a clinical examination to confirm the diagnosis of hypospadias, as a part of routine care. The clinical study investigator will ask to fill out a validated European questionnaire for the exposome and use an occupation/exposure matrix to identify specific atmospheric exposure. A hair sample from the biological mother will be taken for toxicological evaluation of substances accumulated during pregnancy. From the child, a blood sample will be taken for hormonal evaluation of minipuberty and then, another sample in a 5 ml EDTA tube will be taken for DNA collection.; Other: Consultation visit (Visit 1); During visit 1, a pediatric urologist or pediatric endocrinologist will perform a clinical examination to confirm the absence of hypospadias. The clinical study investigator will ask to fill out a validated European questionnaire for the exposome and use an occupation/exposure matrix to identify specific atmospheric exposure. A hair sample from the biological mother will be taken for toxicological evaluation of substances accumulated during pregnancy. From the child, a blood sample will be taken for hormonal evaluation of minipuberty.
Control Group; Biological mother and son without hypospadias	Other: Consultation visit (Visit 1); During visit 1, a pediatric urologist or pediatric endocrinologist will perform a clinical examination to confirm the diagnosis of hypospadias, as a part of routine care. The clinical study investigator will ask to fill out a validated European questionnaire for the exposome and use an occupation/exposure matrix to identify specific atmospheric exposure. A hair sample from the biological mother will be taken for toxicological evaluation of substances accumulated during pregnancy. From the child, a blood sample will be taken for hormonal evaluation of minipuberty and then, another sample in a 5 ml EDTA tube will be taken for DNA collection.; Other: Consultation visit (Visit 1); During visit 1, a pediatric urologist or pediatric endocrinologist will perform a clinical examination to confirm the absence of hypospadias. The clinical study investigator will ask to fill out a validated European questionnaire for the exposome and use an occupation/exposure matrix to identify specific atmospheric exposure. A hair sample from the biological mother will be taken for toxicological evaluation of substances accumulated during pregnancy. From the child, a blood sample will be taken for hormonal evaluation of minipuberty.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Evaluation of toxic exposure via hair sample analysis	An assessment of toxic substance concentrations in maternal hair will evaluate exposure to a wide range of pollutants. The collected hair samples will provide data representing the period from three months before conception to the date of study enrollment. The study will analyze 150 chemical substances and measure their concentrations to assess mothers' exposure to these pollutants during pregnancy.	Day 1 (Visit 1)

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Exposome through validated European questionnaires (QLK4-1999-01422)	Evaluation of parents' environmental exposure (exposome) will be conducted using a simplified version of a validated European questionnaire. This approach aims to elucidate potential differences in exposure between case and control groups.	Day 1 (Visit 1)
Exposome professionnel	Assessment by the job-exposure matrix EDC (Environment disrupting chemicals)	Day 1 (Visit 1)
Pollutant dispersion model	Examination of atmospheric pollutant concentration measurements and their potential impact on maternal exposure during gestation. This study conducts a comparative analysis between two groups: mothers of affected children (case group) and mothers of unaffected children (control group).	Day 1 (Visit 1)
Hormonal analysis by immunoassay principle	Measurement of plasma hormone levels of mini-puberty to investigate a correlation between toxicological dosage and endocrine disruption. FSH, LH, AMH and free Testosterone will be measured by Electrochemiluminescence and Inhibin B by ELISA assay. This assessment of a mini-puberty alteration will be carried out by comparing age-based norms and the values of the control group	Day 1 (Visit 1)

Collaborators and Investigators

• Sponsor: University Hospital, Montpellier
• Investigators: Principal Investigator: Nicolas KALFA, Prof, University Hospital, Montpellier

Publications and helpful links

General Publications

• Skakkebaek NE, Rajpert-De Meyts E, Main KM. Testicular dysgenesis syndrome: an increasingly common developmental disorder with environmental aspects. Hum Reprod. 2001 May;16(5):972-8. doi: 10.1093/humrep/16.5.972.
• Ea V, Bergougnoux A, Philibert P, Servant-Fauconnet N, Faure A, Breaud J, Gaspari L, Sultan C, Paris F, Kalfa N. How Far Should We Explore Hypospadias? Next-generation Sequencing Applied to a Large Cohort of Hypospadiac Patients. Eur Urol. 2021 Apr;79(4):507-515. doi: 10.1016/j.eururo.2020.12.036. Epub 2021 Jan 16.
• Skarin Nordenvall A, Chen Q, Norrby C, Lundholm C, Frisen L, Nordenstrom A, Almqvist C, Nordenskjold A. Fertility in adult men born with hypospadias: A nationwide register-based cohort study on birthrates, the use of assisted reproductive technologies and infertility. Andrology. 2020 Mar;8(2):372-380. doi: 10.1111/andr.12723. Epub 2019 Nov 20.
• Springer A, van den Heijkant M, Baumann S. Worldwide prevalence of hypospadias. J Pediatr Urol. 2016 Jun;12(3):152.e1-7. doi: 10.1016/j.jpurol.2015.12.002. Epub 2015 Dec 31.
• Schneuer FJ, Milne E, Jamieson SE, Pereira G, Hansen M, Barker A, Holland AJA, Bower C, Nassar N. Association between male genital anomalies and adult male reproductive disorders: a population-based data linkage study spanning more than 40 years. Lancet Child Adolesc Health. 2018 Oct;2(10):736-743. doi: 10.1016/S2352-4642(18)30254-2. Epub 2018 Aug 30.
• Asklund C, Jensen TK, Main KM, Sobotka T, Skakkebaek NE, Jorgensen N. Semen quality, reproductive hormones and fertility of men operated for hypospadias. Int J Androl. 2010 Feb;33(1):80-7. doi: 10.1111/j.1365-2605.2009.00957.x. Epub 2009 Mar 5.
• Serrano T, Chevrier C, Multigner L, Cordier S, Jegou B. International geographic correlation study of the prevalence of disorders of male reproductive health. Hum Reprod. 2013 Jul;28(7):1974-86. doi: 10.1093/humrep/det111. Epub 2013 May 12.
• Cortes D, Thorup JM, Visfeldt J. Cryptorchidism: aspects of fertility and neoplasms. A study including data of 1,335 consecutive boys who underwent testicular biopsy simultaneously with surgery for cryptorchidism. Horm Res. 2001;55(1):21-7. doi: 10.1159/000049959.
• Pettersson A, Richiardi L, Nordenskjold A, Kaijser M, Akre O. Age at surgery for undescended testis and risk of testicular cancer. N Engl J Med. 2007 May 3;356(18):1835-41. doi: 10.1056/NEJMoa067588.
• Xing JS, Bai ZM. Is testicular dysgenesis syndrome a genetic, endocrine, or environmental disease, or an unexplained reproductive disorder? Life Sci. 2018 Feb 1;194:120-129. doi: 10.1016/j.lfs.2017.11.039. Epub 2017 Nov 26.
• Olesen IA, Sonne SB, Hoei-Hansen CE, Rajpert-De Meyts E, Skakkebaek NE. Environment, testicular dysgenesis and carcinoma in situ testis. Best Pract Res Clin Endocrinol Metab. 2007 Sep;21(3):462-78. doi: 10.1016/j.beem.2007.04.002.
• Stillman RJ. In utero exposure to diethylstilbestrol: adverse effects on the reproductive tract and reproductive performance and male and female offspring. Am J Obstet Gynecol. 1982 Apr 1;142(7):905-21. doi: 10.1016/s0002-9378(16)32540-6.
• Lymperi S, Giwercman A. Endocrine disruptors and testicular function. Metabolism. 2018 Sep;86:79-90. doi: 10.1016/j.metabol.2018.03.022. Epub 2018 Mar 29.
• Kalfa N, Paris F, Soyer-Gobillard MO, Daures JP, Sultan C. Prevalence of hypospadias in grandsons of women exposed to diethylstilbestrol during pregnancy: a multigenerational national cohort study. Fertil Steril. 2011 Jun 30;95(8):2574-7. doi: 10.1016/j.fertnstert.2011.02.047. Epub 2011 Apr 2.
• Vidaeff AC, Sever LE. In utero exposure to environmental estrogens and male reproductive health: a systematic review of biological and epidemiologic evidence. Reprod Toxicol. 2005 May-Jun;20(1):5-20. doi: 10.1016/j.reprotox.2004.12.015.
• Giwercman A, Rylander L, Lundberg Giwercman Y. Influence of endocrine disruptors on human male fertility. Reprod Biomed Online. 2007 Dec;15(6):633-42. doi: 10.1016/s1472-6483(10)60530-5.
• Sharpe RM, Skakkebaek NE. Testicular dysgenesis syndrome: mechanistic insights and potential new downstream effects. Fertil Steril. 2008 Feb;89(2 Suppl):e33-8. doi: 10.1016/j.fertnstert.2007.12.026.
• Parks LG, Ostby JS, Lambright CR, Abbott BD, Klinefelter GR, Barlow NJ, Gray LE Jr. The plasticizer diethylhexyl phthalate induces malformations by decreasing fetal testosterone synthesis during sexual differentiation in the male rat. Toxicol Sci. 2000 Dec;58(2):339-49. doi: 10.1093/toxsci/58.2.339.
• Sharpe RM. The 'oestrogen hypothesis'- where do we stand now? Int J Androl. 2003 Feb;26(1):2-15. doi: 10.1046/j.1365-2605.2003.00367.x.
• Nilsson EE, Sadler-Riggleman I, Skinner MK. Environmentally induced epigenetic transgenerational inheritance of disease. Environ Epigenet. 2018 Jul 17;4(2):dvy016. doi: 10.1093/eep/dvy016. eCollection 2018 Apr.
• Kalfa N, Paris F, Philibert P, Orsini M, Broussous S, Fauconnet-Servant N, Audran F, Gaspari L, Lehors H, Haddad M, Guys JM, Reynaud R, Alessandrini P, Merrot T, Wagner K, Kurzenne JY, Bastiani F, Breaud J, Valla JS, Lacombe GM, Dobremez E, Zahhaf A, Daures JP, Sultan C. Is Hypospadias Associated with Prenatal Exposure to Endocrine Disruptors? A French Collaborative Controlled Study of a Cohort of 300 Consecutive Children Without Genetic Defect. Eur Urol. 2015 Dec;68(6):1023-30. doi: 10.1016/j.eururo.2015.05.008. Epub 2015 May 23.
• Brouwers MM, van Tongeren M, Hirst AA, Bretveld RW, Roeleveld N. Occupational exposure to potential endocrine disruptors: further development of a job exposure matrix. Occup Environ Med. 2009 Sep;66(9):607-14. doi: 10.1136/oem.2008.042184. Epub 2009 Mar 13.
• Beranger R, Hardy EM, Dexet C, Guldner L, Zaros C, Nougadere A, Metten MA, Chevrier C, Appenzeller BMR. Multiple pesticide analysis in hair samples of pregnant French women: Results from the ELFE national birth cohort. Environ Int. 2018 Nov;120:43-53. doi: 10.1016/j.envint.2018.07.023. Epub 2018 Jul 29.
• Peng FJ, Hardy EM, Beranger R, Mezzache S, Bourokba N, Bastien P, Li J, Zaros C, Chevrier C, Palazzi P, Soeur J, Appenzeller BMR. Human exposure to PCBs, PBDEs and bisphenols revealed by hair analysis: A comparison between two adult female populations in China and France. Environ Pollut. 2020 Dec;267:115425. doi: 10.1016/j.envpol.2020.115425. Epub 2020 Aug 15.
• Davis CE Jr, Wiley WB, Faulconer RJ. Necrosis of the female breast complicating oral anticoagulant treatment. Ann Surg. 1972 May;175(5):647-56. doi: 10.1097/00000658-197205000-00004. No abstract available.

Study record dates

Study Major Dates

• Study Start (Actual): October 3, 2024
• Primary Completion (Estimated): September 30, 2027
• Study Completion (Estimated): October 3, 2027

Study Registration Dates

• First Submitted: September 25, 2024
• First Submitted That Met QC Criteria: October 2, 2024
• First Posted (Actual): October 8, 2024

Study Record Updates

• Last Update Posted (Actual): November 26, 2025
• Last Update Submitted That Met QC Criteria: November 20, 2025
• Last Verified: November 1, 2025

More Information

Terms related to this study

• Keywords: Environment; Birth defect; Exposome; Endocrine-disrupting chemicals; Hormones minipuberty; Toxicology hair
• Additional Relevant MeSH Terms: Urogenital Diseases; Genital Diseases; Genital Diseases, Male; Male Urogenital Diseases; Female Urogenital Diseases; Female Urogenital Diseases and Pregnancy Complications; Urogenital Abnormalities; Penile Diseases; Congenital, Hereditary, and Neonatal Diseases and Abnormalities; Congenital Abnormalities; Hypospadias
• Other Study ID Numbers: RECHMPL22_0559

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No
• product manufactured in and exported from the U.S.: No