Effect of chronic low-dose phthalate exposure on the immature primate testis

长期低剂量邻苯二甲酸盐暴露对未成熟灵长类动物睾丸的影响

• 批准号: 7530079
• 负责人: INA DOBRINSKI
• 金额: $ 20.99万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-08 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7530079
• 关键词: Adolescent; Affect; Animals; Attention; Biological Models; Catheterization; Cell Differentiation process; Cell Maturation; Cell physiology; Childhood; Chronic; Data; Development; Dibutyl Phthalate; Diethylhexyl Phthalate; Dose; Endocrine; Environment; Environmental Exposure; Epidemiologic Studies; Esters; Experimental Designs; Exposure to; Fertility; Food; Future; Gene Expression; Gene Expression Profiling; Genes; Genome; Germ Cells; Gonadotropins; Health; Human; Incidence; Infant; Intravenous; Investigation; Lead; Link; Long-Term Effects; Macaca; Macaca mulatta; Male Genital Organs; Metabolism; Microarray Analysis; Modeling; Molecular; Monkeys; Mus; Organ; Pathway interactions; Patients; Pattern; Perinatal Exposure; Phthalic Acids; Plasticizers; Plastics; Primates; Public Health; RNA; Rattus; Research; Research Design; Research Project Grants; Research Proposals; Reverse Transcriptase Polymerase Chain Reaction; Rodent; Rodent Model; Self Care; Somatic Cell; Steroid biosynthesis; System; Testicular Neoplasms; Testing; Testis; Time; Tissue Donors; Tissue Grafts; Tissue-Specific Gene Expression; Tissues; Toxic effect; Toxicology; Toy; Work; Xenograft Model; Xenograft procedure; age effect; base; consumer product; cost; dosage; exposed human population; human male; insight; leydig interstitial cell; male; medical supply; mono-(2-ethylhexyl)phthalate; nonhuman primate; novel; phthalate; phthalates; phthalic acid; reproductive; reproductive development; sertoli cell; toxicant

DESCRIPTION (provided by applicant): The objective of this exploratory research project is to study the effects of chronic exposure to phthalate esters on the infant primate testis in a novel xenograft model system. Environmental exposure to reproductive toxicants is of serious concern to human health and has been implicated in both the apparent decline of male fertility and a rise in testicular neoplasms. Phthalate esters are widely used as plasticizers in consumer products and can leach into the environment, resulting in significant human exposure, especially during childhood and in pediatric patients requiring long-term catheterization or intravenous treatments. Di-(2- ethylhexyl) phthalate (DEHP) is the most ubiquitous phthalate and its metabolite MEHP is a known testicular toxicant. Di-n-butyl phthalate (DBP) is commonly found in personal care products and fetal exposure to DBP affects male reproductive tract development in rats. Historically, research into the reproductive toxicology of phthalates has been evaluated following gestational exposure in rodents, but species-specific differences in metabolism and sensitivity of the reproductive organs suggest that results obtained in rodent models may not extrapolate well to humans. As epidemiological studies link phthalate exposure during childhood to disturbances of human male reproductive development, there is a clear need for studies in primate models, as emphasized by the NTP-CERHR Expert Panel on the Reproductive and Developmental Toxicity of DEHP. In this revised exploratory project we propose to utilize a novel primate testis tissue xenografting system developed in our lab to study the effect of DEHP and DBP on testicular maturation and testis function in immature rhesus monkey testes. Specific aim 1 will test the hypothesis that exposure to environmentally relevant concentrations of DEHP or DBP will adversely affect testicular somatic cell function and germ cell development. Specifically, testis tissue from infant and juvenile monkeys will be ectopically grafted to mouse hosts and exposed to various doses of DEHP or DBP for 3 months. At various time points, Sertoli cell maturation, steroidogenic function of Leydig cells, and exogenous gonadotropin-induced germ cell differentiation will be compared in treated and control tissue to assess the potential effects of chronic phthalate exposure. Specific aim 2 will investigate the mechanisms underlying phthalate toxicity in the infant primate testis. Phthalate-induced changes in global gene expression patterns in testis tissue from infant monkeys will be evaluated by microarray analysis and confirmed by RT-PCR. Special attention will be given to differential expression of genes involved in steroidogenesis. Successful completion of this exploratory project will provide insights into the sensitivity to phthalate esters, characterize effects on testicular somatic cell maturation, and begin to elucidate the molecular mechanisms underlying phthalate toxicity in the immature primate testis. Results from the proposed project will provide the basis for future studies performed in the developing primate. PUBLIC HEALTH RELEVANCE: Rodent models are conventionally utilized to study the potential effects of suspected reproductive toxicants, such as phthalates, on the immature human testis, but species-specific differences in metabolism and sensitivity of the reproductive organs may not make them ideal model systems. While studies in primates are more likely applicable to the human, they are often cost-prohibitive and ethically restrictive. In this research proposal we describe a novel system in which pieces of primate testis tissue are grafted into mice to investigate the mechanisms of reproductive phthalate toxicity in primate testis without extensive experimentation in primates.

描述（由申请人提供）：该探索性研究项目的目的是在新型异种移植模型系统中研究长期接触邻苯二甲酸酯对婴儿灵长类动物睾丸的影响。环境中生殖毒物的暴露严重影响人类健康，并与男性生育能力的明显下降和睾丸肿瘤的增加有关。邻苯二甲酸酯广泛用作消费品中的增塑剂，可渗入环境中，导致人体大量接触邻苯二甲酸酯，特别是在儿童期和需要长期导尿或静脉治疗的儿科患者中。邻苯二甲酸二(2-乙基己基)酯 (DEHP) 是最普遍存在的邻苯二甲酸酯，其代谢物 MEHP 是一种已知的睾丸毒物。邻苯二甲酸二正丁酯 (DBP) 常见于个人护理产品中，胎儿接触 DBP 会影响大鼠的雄性生殖道发育。历史上，对邻苯二甲酸盐生殖毒理学的研究是在啮齿动物妊娠期暴露后进行评估的，但生殖器官代谢和敏感性的物种特异性差异表明，在啮齿动物模型中获得的结果可能无法很好地推断到人类。由于流行病学研究将儿童时期接触邻苯二甲酸盐与人类男性生殖发育障碍联系起来，因此显然需要在灵长类动物模型中进行研究，正如 NTP-CERHR DEHP 生殖和发育毒性专家小组所强调的那样。在这个修订后的探索性项目中，我们建议利用我们实验室开发的新型灵长类睾丸组织异种移植系统来研究 DEHP 和 DBP 对未成熟恒河猴睾丸睾丸成熟和睾丸功能的影响。具体目标 1 将检验以下假设：暴露于环境相关浓度的 DEHP 或 DBP 会对睾丸体细胞功能和生殖细胞发育产生不利影响。具体来说，将来自幼猴和幼猴的睾丸组织异位移植到小鼠宿主上，并暴露于不同剂量的DEHP或DBP 3个月。在不同的时间点，将在处理组织和对照组织中比较支持细胞的成熟、间质细胞的类固醇生成功能和外源性促性腺激素诱导的生殖细胞分化，以评估慢性邻苯二甲酸盐暴露的潜在影响。具体目标 2 将研究婴儿灵长类动物睾丸中邻苯二甲酸盐毒性的机制。邻苯二甲酸盐诱导的幼猴睾丸组织中整体基因表达模式的变化将通过微阵列分析进行评估，并通过 RT-PCR 进行确认。将特别关注类固醇生成相关基因的差异表达。该探索性项目的成功完成将深入了解邻苯二甲酸酯的敏感性，表征对睾丸体细胞成熟的影响，并开始阐明未成熟灵长类睾丸中邻苯二甲酸酯毒性的分子机制。拟议项目的结果将为未来在发育中的灵长类动物中进行的研究提供基础。公共卫生相关性：啮齿动物模型通常用于研究可疑生殖毒物（例如邻苯二甲酸盐）对未成熟人类睾丸的潜在影响，但生殖器官代谢和敏感性的物种特异性差异可能无法使它们成为理想的模型系统。虽然灵长类动物的研究更有可能适用于人类，但它们往往成本高昂且受到伦理限制。在这项研究计划中，我们描述了一种新的系统，其中将灵长类动物睾丸组织移植到小鼠体内，以研究灵长类动物睾丸中邻苯二甲酸盐生殖毒性的机制，而无需在灵长类动物中进行大量实验。