Mechanisms of ovarian endocrine disruption at single-cell resolution

单细胞分辨率的卵巢内分泌干扰机制

基本信息

批准号：
10681228
负责人：
Jennifer Monique Cossaboon
金额：
$ 4.03万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10681228
关键词：
Adult Binding Biological Assay Biological Markers Breast Cancer Risk Factor Breeding Cell Differentiation process Cells Chemical Exposure Chemicals Country Data Daughter Development Diethylstilbestrol Disease Dose Economics Embryo Endocrine Disruptors Endocrine System Diseases Endocrine disruption Environment Environmental Estrogen Environmental Exposure Enzymes Estrogen Receptors Estrogens Ethics Experimental Models Exposure to Female Fertility Fertilization Fishes Gene Expression Regulation Genes Genetic Genetic Transcription Genome Gonadal Steroid Hormones Gonadal structure Health Histology Hormone Receptor Hormones Human Hypothalamic structure Impairment Infertility Isomerism Japanese Killifish Knowledge Larva Life Life Cycle Stages Long-Term Effects Longevity Mammals Masks Maternal Exposure Measures Mediating Metabolism Modeling Molecular Nature Nuclear Hormone Receptors Onset of illness Oocytes Oryziinae Ovarian Ovarian Diseases Ovarian Follicle Ovary Parents Pathway interactions Pesticides Physiological Pituitary Gland Poison Population Pregnancy Rate Production Public Health Research Resolution Risk Role Sequence Homology Signal Transduction Somatic Cell Sorting Steroid biosynthesis Testing Toxicology Woman Work Zebrafish analytical tool androgenic cancer risk cell type contaminated water dichlorodiphenyltrichloroethane egg estrogenic experimental study exposed human population female fertility fetal gene regulatory network genotypic sex gonad development high risk hormonal signals in utero in vivo in vivo Model individual response innovation insight lipophilicity novel oocyte maturation organ growth organochlorine pesticide prenatal exposure reproductive reproductive system disorder reproductive system neoplasm screening sex sex determination sexual dimorphism single cell mRNA sequencing steroid hormone transcriptome sequencing transcriptomics translational model xenoestrogen

项目摘要

Mechanisms of Ovarian Endocrine Disruption at Single-Cell Resolution Endocrine disrupting chemicals (EDCs) are exogenous chemicals that interfere with endogenous hormone synthesis, metabolism and signaling. EDC exposure during early gonadal development is suspected to play a role in ovarian dysgenesis later in life. Advancing understanding of endocrine disruption at the molecular and cellular level is essential for determining possible developmental origins of adult-onset reproductive disease. This demands sensitive in vivo models with short lifespans and innovative analytical tools to detect genetic perturbations induced by chemical exposures. The Japanese medaka fish (Oryzias latipes) is among the most studied experimental models used in EDC screening for developmental and reproductive effects. Not only is the medaka genome extensively annotated compared to other fish models, but it shares chromosomal sex determination, hormone receptor sequence homology, and sex hormone signaling axes with humans. Exposure to estrogen-contaminated water during ovarian development causes significantly delayed oocyte maturation and reduced egg production in adult medaka. However, specific mechanisms by which heterogenous cell populations of the developing ovary respond to estrogenic chemicals remain unclear. This proposal seeks to investigate how early developmental exposures to xenoestrogens alter gene regulation networks in differentiating ovarian cells, and which long-term transcriptional changes are associated with decreased female fertility. My working hypothesis is that early life stage exposures to estrogenic DDTs induce long-term transcriptional effects in ovarian somatic gonad cells, reducing fertility later in life. I will investigate this hypothesis by exposing medaka to environmentally relevant levels of o,p’- dichlorodiethyltrichloroethane (o,p’-DDT), an estrogenic isomer of the legacy organochlorine pesticide that remains a persistent public health concern. Exposures will occur during a key window of ovarian development to define organizational effects at the functional, cellular, and transcriptional levels. Aim 1 will define the long-term reproductive consequences of o,p’-DDT exposures during an early window of ovarian differentiation using functional breeding assessments and targeted quantification of estrogen- and steroidogenesis-related genes. Aim 2 will use single cell transcriptomics (scRNA-seq) to gain a comprehensive view of which ovarian somatic cell types and gene regulatory networks (both steroid hormone- dependent and hormone-independent), are permanently altered by early stage o,p’-DDT exposure. The proposed work will elucidate lasting functional and molecular changes induced by xenoestrogens during ovarian differentiation and follicular assembly in medaka, which aligns with fetal ovarian development in humans. Comprehensive identification of DDT-inducible genes may reveal novel exposure biomarkers and plausible mechanisms of endocrine and reproductive diseases.

单细胞分辨率的卵巢内分泌干扰机制内分泌干扰化学物质（EDC）是干扰内源激素的外源化学物质早期性腺发育过程中 EDC 的合成、代谢和信号传导被怀疑发挥着重要作用。促进对内分泌干扰的分子和机制的理解。细胞水平对于确定成人发病的生殖疾病的可能发育起源至关重要。这需要寿命短的敏感体内模型和创新的分析工具来检测遗传日本青鳉鱼（Oryzias latipes）是由化学物质暴露引起的干扰最严重的鱼之一。研究用于 EDC 筛选的实验模型不仅是对发育和生殖的影响。与其他鱼类模型相比，青鳉基因组得到了广泛注释，但它具有相同的染色体性别测定、激素受体序列同源性以及与人类的性激素信号轴。卵巢发育过程中接触雌激素污染的水会导致卵母细胞显着延迟然而，成年青鳉成熟和产蛋量减少的具体机制。发育中卵巢的异质细胞群对雌激素化学物质的反应仍不清楚。该提案旨在研究早期发育暴露于异雌激素如何改变基因调控分化卵巢细胞的网络，以及与长期转录变化相关的我的工作假设是生命早期接触雌激素滴滴涕。诱导卵巢体细胞性腺细胞的长期转录效应，降低晚年的生育能力。将通过将青鳉暴露于环境相关水平的 O,P'- 来研究这一假设二氯二乙基三氯乙烷 (o,p’-DDT)，是传统有机氯农药的雌激素异构体，仍然是一个持续存在的公共卫生问题，暴露发生在卵巢发育的关键时期。定义功能、细胞和转录水平的组织效应。目标 1 将确定早期窗口期间 o,p'-DDT 暴露的长期生殖后果使用功能育种评估和雌激素和雌激素的目标定量进行卵巢分化目标 2 将使用单细胞转录组学 (scRNA-seq) 来获得类固醇生成相关基因。全面了解卵巢体细胞类型和基因调控网络（类固醇激素- 依赖性和激素非依赖性），会因早期 o,p'-DDT 暴露而永久改变。拟议的工作将阐明异雌激素在青鳉的卵巢分化和卵泡组装，与胎儿卵巢发育一致人类 DDT 诱导基因的全面鉴定可能会揭示新的暴露生物标志物和内分泌和生殖疾病的合理机制。