# Male preconception phthalates and offspring embryo and sperm allele-specific methylome programming

> **NIH NIH R01** · UNIVERSITY OF MASSACHUSETTS AMHERST · 2020 · $446,729

## Abstract

Project Summary
Phthalates, a class of endocrine disrupting compounds (EDCs) used in plastics and personal care products,
are ubiquitous environmental contaminants resulting in widespread human exposure. Epidemiologic data
implicate paternal phthalates with adverse reproductive health including poor sperm quality, and more recently,
with longer time to pregnancy − the latter suggests a sperm-derived effect. A growing body of compelling data
demonstrates that environmental exposures can be embodied within the developing male germ cell through
epigenetic marks, which in turn, can impart information at fertilization to affect the trajectory of health and
development of offspring. It is known that prenatal epigenetic reprogramming of male germ cells is a
particularly susceptible window to environmental exposures such as phthalates. Research has now emerged
demonstrating that other such susceptible windows exist during germ cell development. Our objective is to
determine the effects of adult exposure to
di(2-ethylhexyl) phthalate (DEHP), di-n-butyl phthalate (DBP) and
the mixture of DEHP and DBP on sperm DNA methylation and the
 persistence of sperm-derived changes on
DNA methylation programming of the conceptus and F1 adult germ cells. We will exposure adult reproductive
age male C57BL/6J mice with 0, 2.5 mg/kg/day of DEHP, 2.5 mg/kg/day of DBP, and its mixture (DEHP and
DBP each at 2.5 mg/kg/day) for 70 days – approximately two spermatogenesis cycles. Epididymal sperm will
be collected from mice in each group and will be analyzed for whole genome bisulfite sequencing (WGBS).
Next, exposed C57BL/6J males will be mated with unexposed DBA/2Jfemales, which will allow for allele-
specific analysis across the genome in the F1 hybrid embryos produced. We will perform RNA-seq and WGBS
in embryonic and placental cells of gastrulating E6.5 embryos to determine the effects of paternal DEHP and/or
DBP on each distinct lineage. Finally, we will analyze sperm quality parameters, oocyte follicle staging and
reproductive hormones in male and female F1 offspring. We will also perform RNA-seq and WGBS on F1
sperm and oocytes. This research proposed is expected to identify an epigenetic understanding via sperm
methylation of the effects of preconception male EDC exposure and offspring reproductive health.
Understanding the involvement of sperm epigenetics within the exposure-disease paradigm will inform
avenues of translational research for the development of novel approaches for the treatment and prevention of
adverse reproductive health.

## Key facts

- **NIH application ID:** 9996701
- **Project number:** 5R01ES028214-04
- **Recipient organization:** UNIVERSITY OF MASSACHUSETTS AMHERST
- **Principal Investigator:** J. Richard Pilsner
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $446,729
- **Award type:** 5
- **Project period:** 2017-09-01 → 2021-03-01

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9996701

## Citation

> US National Institutes of Health, RePORTER application 9996701, Male preconception phthalates and offspring embryo and sperm allele-specific methylome programming (5R01ES028214-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9996701. Licensed CC0.

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