Clinical trial NCT03679000: The Effect of Periconceptional Phthalates Exposure on Couples' Reproductive Health
Official title: Periconceptional Phthalates Exposure and Its Epigenetic Effect on Fertility, Embryo Development, and Neonatal Outcomes: a Prospective Study
Study type: OBSERVATIONAL
Overall status: UNKNOWN
Conditions: Environmental Exposure
Infertility
Keywords: environmental exposure
epigenetic process
embryo development
infertility
cohort study
Lead sponsor: Tongji Hospital
Sponsor class: OTHER
First posted: 2018-09-20
Enrollment: 2122
Sex: ALL
Minimum age (years): 20
Maximum age (years): 45
Standard ages: ADULT
Accepts healthy volunteers: no
Observational model: COHORT
Time perspective: PROSPECTIVE
Sampling method: NON_PROBABILITY_SAMPLE
Study population: The study population is a group of childbearing couples who are seeking assisted reproductive technologies for having a baby in the reproductive center in Tongji Hospital.

Brief summary: The study is a prospective cohort study, which is aiming to explore periconceptional phthalates exposure and its potential epigenetic effect on fertility, embryo development, and neonatal outcomes.

Detailed description: The study is a prospective cohort study, and its subjects are childbearing couples who are seeking assisted reproductive technologies for having a baby in the reproductive center in Tongji Hospital.

1. All enrolled couples will be needed to sign an informed consent and complete a questionnaire for collecting information about demographics, life-style, and medical and reproductive histories.
2. Biological samples will be collected from the participants to measure phthalates metabolites in order to evaluate the exposure of single person or the couple in different time phase.The samples include urine, blood that will be left after the hormones measurement for both spouses. The follicular fluid will be obtained from females on the day of oocyte retrieval. The left sperm samples after sperm analysis will be obtained from men. In the reproductive center of Tongji Hospital, hormone measurement for women and sperm analysis for men are required for the In vitro fertilization and embryo transfer in order to evaluate the fertility of couples.And the blastocyst culture medium will be obtained from couples after blastocyst transfer. All samples will be stored at -80℃ until further analysis.
3. Measurements of phthalates metabolites in the biological samples.
4. Collecting data from electronic medical records and follow-up.The basic information, clinical data and reproductive outcomes of the subjects will be collected by reviewing the electronic medical charts, including the height, weight, basic hormone levels, stimulation protocol and time, insemination protocol, fertilization rate, cleavage rate, excellent embryo rate, blastocyst formation rate, clinical pregnancy rate, as well as live birth rate，the newborns' height, weight, and gestational age, etc.
5. Analyze the miRNA expression profiles of blastocyst culture medium.
6. Statistical analysis. The investigators will analyze the association between the couples' phthalates exposure levels and the reproductive outcomes, and explore the association among periconceptional phthalates exposure, epigenetic changes and reproductive outcomes.

Eligibility criteria: 一. Inclusion and exclusion criteria for primary study:

The exclusion of men:

1. Occupational exposures
2. Couples living in separate places
3. Congenital diseases

   * Congenital azoospermia
   * Ejaculation dysfunction
   * Family history of genetic diseases
4. Acquired diseases

   * Genitourinary infections
   * Mumps
   * Endocrine diseases
   * Sexually transmitted diseases
   * Hyperthyroidism
   * Liver failure
   * Renal failure
   * Neurological diseases
   * Paraplegia
   * Acquired testicular injury
5. Patient who have received surgeries：

   * Inguinal hernia surgery
   * Vasectomy
   * Bladder neck surgery
   * Pelvic surgery
6. Patients who have used antitumor drugs and hormone

The exclusion of women:

1. Occupational exposures
2. Couples living in separate places
3. Congenital gonadal dysplasia

   * Genital malformation
   * Chromosomal abnormalities
   * Family history of genetic diseases
4. Endocrine and metabolic diseases

   * Diabetes
   * Hyperprolactinemia
   * Thyroid and adrenal diseases
5. Cardiovascular diseases

   * Dyslipidemia
   * Systemic lupus erythematosus
   * Other rheumatic immune diseases
6. Patients who have received the following treatment:

   * Ovarian surgery
   * Thyroid surgery
   * Pituitary surgery
   * Anti-tumor radiotherapy
   * Chemotherapy
7. Patients with previous history as followed:

   * Endometrial injury
   * Endometrial polyps
   * Endometritis
   * Uterine fibroids
   * Endometriosis
8. Tubal effusion

二. Inclusive and exclusive criteria for blastocyst culture medium collection at the later stage.

Inclusion criteria:

1. Subjects in the primary part of the study
2. ·Normal gender of men and women

   * No genetic diseases or congenital diseases
   * No family genetic history
3. Normal sperm analysis for men
4. Female age \<35 years
5. ·Normal Ovarian function for women

   * Normal endocrine function for women
   * No uterine disease or abnormality
6. The causes of infertility :

   * Tubal factors
   * Unexplained
7. The protocol of controlled ovary hyperstimulation :

   * A routine long-term protocol

Exclusion criteria:

1. Subjects with smoking or alcohol abuse in the past 3 months
2. Patients with overweight or obesity (BMI ≥ 25)
3. Embryos with chromosomal abnormalities confirmed by PGS

Primary outcomes:
measure: Phthalates exposure levels
timeFrame: 12/2021
Secondary outcomes:
measure: Exposure characteristics of phthalates
timeFrame: 12/2021
measure: Reproductive outcomes
timeFrame: 12/2021
measure: The expression profiles of miRNA
timeFrame: 12/2021
Other outcomes:
Locations:
city: Wuhan
country: China
facility: Tongji Hospital
state: Hubei
Study officials:
affiliation: Tongji Hospital
name: Yufeng Li, Doctor
role: PRINCIPAL_INVESTIGATOR
References:
citation: Du YY, Fang YL, Wang YX, Zeng Q, Guo N, Zhao H, Li YF. Follicular fluid and urinary concentrations of phthalate metabolites among infertile women and associations with in vitro fertilization parameters. Reprod Toxicol. 2016 Jun;61:142-50. doi: 10.1016/j.reprotox.2016.04.005. Epub 2016 Apr 8.
citation: Wang YX, Zeng Q, Sun Y, You L, Wang P, Li M, Yang P, Li J, Huang Z, Wang C, Li S, Dan Y, Li YF, Lu WQ. Phthalate exposure in association with serum hormone levels, sperm DNA damage and spermatozoa apoptosis: A cross-sectional study in China. Environ Res. 2016 Oct;150:557-565. doi: 10.1016/j.envres.2015.11.023. Epub 2015 Dec 3.
citation: Wang YX, You L, Zeng Q, Sun Y, Huang YH, Wang C, Wang P, Cao WC, Yang P, Li YF, Lu WQ. Phthalate exposure and human semen quality: Results from an infertility clinic in China. Environ Res. 2015 Oct;142:1-9. doi: 10.1016/j.envres.2015.06.010. Epub 2015 Jun 15.
citation: Zeng Q, Chen YZ, Xu L, Chen HX, Luo Y, Li M, Yue J, Liu AL, Li YF, Lu WQ. Evaluation of exposure to trihalomethanes in tap water and semen quality: a prospective study in Wuhan, China. Reprod Toxicol. 2014 Jul;46:56-63. doi: 10.1016/j.reprotox.2014.03.005. Epub 2014 Mar 13.
citation: Zeng Q, Li M, Xie SH, Gu LJ, Yue J, Cao WC, Zheng D, Liu AL, Li YF, Lu WQ. Baseline blood trihalomethanes, semen parameters and serum total testosterone: a cross-sectional study in China. Environ Int. 2013 Apr;54:134-40. doi: 10.1016/j.envint.2013.01.016. Epub 2013 Feb 26.