In Vitro Analysis of the Effects of Acute and Chronic Phthalate Exposures on Leydig Cell Testosterone Production, and the Molecular Mechanisms Involved

急性和慢性邻苯二甲酸盐暴露对间质细胞睾酮产生的影响以及所涉及的分子机制的体外分析

• 批准号: 10046384
• 负责人: Kassim Traore
• 金额: $ 31.25万
• 依托单位: CAMPBELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10046384
• 关键词: Acute; Address; Adult; Affect; Animals; Area; Award; Basic Science; Cell Differentiation process; Cell Proliferation; Cells; Chronic; Cities; Clinical Research; Collaborations; Cryptorchidism; Defect; Development; Diethylhexyl Phthalate; Dose; Elements; Endocrine Disruptors; Environment; Equilibrium; Equipment; Exposure to; Faculty; Funding; Gene Expression; Grant; Hormone Responsive; Hormones; Human; Human Resources; Hypospadias; In Vitro; Industrialization; Ingestion; Inhalation; Institution; Laboratories; Long-Term Effects; Luteinizing Hormone; Malignant neoplasm of testis; Medical Students; Mentorship; Methods; Molecular; Morphogenesis; North Carolina; Organ; Oxidation-Reduction; Oxidative Stress; Pathway interactions; Plasticizers; Positioning Attribute; Predisposition; Pregnancy; Production; Proliferating; Public Health Schools; Reactive Oxygen Species; Reporting; Reproduction; Reproductive system; Research; Research Project Grants; Risk Factors; Seminal fluid; Seminiferous tubule structure; Stem Cell Development; Steroids; Students; Surface; System; Testosterone; Universities; career; dermal exposure; graduate student; in vivo; innovation; interest; leydig interstitial cell; medical schools; member; mono-(2-ethylhexyl)phthalate; phthalates; protein expression; response; rural underserved; sex; stem; stem cells; undergraduate student; university student

By interfering with hormone function and action, endocrine-disrupting chemicals (EDCs) such as the plasticizer di-(2-ethylhexyl) phthalate (DEHP) can have negative effects on development and reproduction. With the support of a Research Enhancement (SC2) Award, when he was a faculty member at Elizabeth City State University, Dr. Traore found that in vivo exposure to DEHP during gestation had negative effects on testosterone (T) production in the adult, and that early exposure exacerbated the effects of subsequent exposures. Now at Campbell University School of Medicine, Dr. Traore is proposing to conduct in vitro mechanistic studies of both acute and chronic DEHP effects on T production by Leydig cells. Until recently, Leydig cells isolated and cultured by available methods have been able to produce and maintain high levels of T production for only 3-5 days. Therefore, it has not yet been feasible to use primary cells in vitro to examine the direct, long-term effects of environmentally relevant concentrations of EDCs or to critically evaluate mechanisms involved in their actions on Leydig cells. However, Dr. Zirkin’s lab at the Johns Hopkins Bloomberg School of Public Health (JHSPH) has recently developed a method by which Leydig cells derived from the differentiation of Leydig stem cells are able to produce and sustain high levels of T production for longer periods, several months. Dr. Zirkin, who served in a mentorship position for the SC2 Award, is continuing to collaborate with Dr. Traore and his students. The availability of primary Leydig cells able to sustain high levels of T production will be of great value for our proposed studies of the chronic effects of environmentally relevant phthalate concentrations under highly controlled conditions, and the mechanisms by which they function. With the availability of these cells, we will conduct in vitro studies of the direct effects of MEHP on Leydig cell formation from stem cells, the effects of MEHP exposure on the adult Leydig cells, and the effects of early exposures on the consequences of subsequent exposures. We also will conduct mechanistic studies of acute and long-term effects of phthalate-induced oxidative stress on T production. Campbell University has the equipment and facilities necessary for the proposed studies, as well as outstanding medical students, graduate students and undergraduates who will be involved in this research project. With funds of this REAP grant, Dr. Traore will be able to support and expand ongoing research in his lab at Campbell University, an institution located in a rural, underserved area of North Carolina. The extensive collaboration between the Johns Hopkins School of Public Health and Campbell University will not only help excite Campbell University students but also increase their interest in considering careers in basic science or clinical research. Moreover, the project undoubtedly will attract other Campbell University faculty to seek funding to develop their research, and help strengthen the research environment on campus.

增塑剂邻苯二甲酸二（2-乙基己基）酯 (DEHP) 等内分泌干扰化学物质 (EDC) 会干扰激素功能和作用，从而对发育和生殖产生负面影响。 Traore 博士在伊丽莎白城市州立大学任教时发现，妊娠期间体内暴露于 DEHP 对成人睾酮 (T) 的产生产生负面影响，而且早期暴露加剧了后续暴露的影响。现在，在坎贝尔大学医学院，Traore 博士建议对 Leydig 细胞的急性和慢性 DEHP 影响进行体外机制研究，直到最近，Leydig 细胞才被分离和培养。方法只能产生并维持高水平的 T 产生仅 3-5 天，因此，在体外使用原代细胞来检查环境相关浓度的 EDC 或 EDC 的直接、长期影响尚不可行。批判地然而，Zirkin 博士在约翰霍普金斯大学的实验室评估了它们对 Leydig 细胞的作用机制。 布隆伯格公共卫生学院 (JHSPH) 最近开发了一种方法，由 Leydig 干细胞分化而来的 Leydig 细胞能够产生并维持高水平的 T 产生较长时间，Zirkin 博士曾在该学院任职。 SC2 奖的导师职位，正在继续与 Traore 博士和他的学生合作，能够维持高水平 T 产生的原代间质细胞的可用性对于我们拟议的环境相关慢性影响的研究具有重要价值。高度控制条件下的邻苯二甲酸盐浓度及其发挥作用的机制随着这些细胞的可用性，我们将在体外研究 MEHP 对干细胞形成 Leydig 细胞的直接影响，以及 MEHP 暴露对成人的影响。间质细胞，以及早期暴露对后续暴露后果的影响我们还将针对邻苯二甲酸盐诱导的氧化应激对 T 细胞的急性和长期影响进行机制研究。 坎贝尔大学拥有拟议研究以及生产所需的设备和设施。 参与该研究项目的优秀医学生、研究生和本科生将能够利用 REAP 资助的资金，支持和扩大他在坎贝尔大学（位于农村地区的机构）实验室正在进行的研究。约翰·霍普金斯大学公共卫生学院和坎贝尔大学之间的广泛合作不仅有助于激发坎贝尔大学学生的兴趣，而且还能增加他们考虑从事基础科学或临床研究职业的兴趣。将吸引其他坎贝尔大学教师寻求资金来发展他们的研究，并帮助加强校园的研究环境。