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.

通过干扰激素的功能和作用，内分泌干扰化学物质（EDCs），如增塑剂邻苯二甲酸二（2-乙基己基）酯（DEHP）会对发育和繁殖产生负面影响。特拉奥雷博士在伊丽莎白城市州立大学担任教职时，获得了一项研究增强奖（SC2）的支持，他发现妊娠期体内暴露于DEHP对成年后睾酮(T)的产生有负面影响，并且早期暴露加剧了随后暴露的影响。现在，在坎贝尔大学医学院，特拉奥雷博士提议进行急性和慢性DEHP对间质细胞产生T的影响的体外机制研究。直到最近，通过现有方法分离和培养的间质细胞只能产生并维持3-5天的高水平T分泌。因此，在体外使用原代细胞来检查环境相关浓度的EDCs的直接、长期影响或批判性地评估其对间质细胞作用的机制尚不可行。然而，Zirkin博士在约翰霍普金斯大学的实验室