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.

单细胞分辨率时卵巢内分泌干扰的机制内分泌干扰化学物质（EDC）是干扰内源恐怖剂的外源化学物质合成，代谢和信号传导。怀疑性腺早期开发期间的EDC暴露会发挥在生命之后的卵巢失调中的作用。促进对分子内分泌干扰的理解和细胞水平对于确定成人发作复制疾病的可能发育起源至关重要。这需要具有短寿命和创新分析工具的敏感体内模型，以检测遗传化学暴露引起的扰动。日本的Medaka鱼（Oryzias latipes）是最多的用于EDC筛选的研究烟熏实验模型，以进行发育和生殖效应。不仅是与其他鱼类模型相比，Medaka基因组广泛注释，但共享染色体性别确定，马对马对序列同源性和与人类的性马信号传导轴。卵巢发育过程中接触雌激素污染的水会显着延迟卵母细胞成人Medaka的成熟和卵产量减少。但是，特定机制发育中的卵巢的异质细胞群对雌激素化学物质反应尚不清楚。该提案旨在调查异种雌激素的早期发育暴露如何改变基因调节区分卵巢细胞的网络，以及长期转录变化与女性生育能力降低。我的工作假设是，早期生命阶段暴露于雌激素DDTS 诱导卵巢体细胞细胞的长期转录作用，从而降低生命的生育能力。将通过将MEDAKA暴露于与环境相关的O，P’-的水平来调查这一假设二氯二甲基三氯乙烷（O，p’-DDT），旧的有机氯农药的雌激素异构体仍然是一个持续的公共卫生问题。暴露将在卵巢开发的关键窗口中发生在功能，细胞和转录水平上定义组织效应。 AIM 1将定义O，P’-DDT暴露的长期生殖后果使用功能育种评估以及雌激素和靶向定量的卵巢分化类固醇发生相关的基因。 AIM 2将使用单细胞转录组学（SCRNA-SEQ）获得综合视图哪种卵巢体细胞类型和基因调节网络（均类固醇骑马 - 依赖和独立于马的），通过早期O，P’-DDT暴露会永久改变。所提出的工作将阐明异雌激素引起的持久功能和分子变化。 Medaka中的卵巢分化和卵泡组件，与胎儿卵巢发育保持一致人类。全面鉴定DDT诱导基因可能揭示了新型的暴露生物标志物，并且内分泌和再现疾病的合理机制。