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

标题：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: 10730350
• 负责人: Kassim Traore
• 金额: $ 40.19万
• 依托单位: DUQUESNE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10730350
• 关键词: Acute; Address; Adult; Affect; Amniotic Fluid; Animals; Antioxidants; Attention; Basic Science; CCL21 gene; Cell Line; Cell Separation; Cells; Chemicals; Cholesterol; Chronic; Clinical Research; Cosmetics; Cryptorchidism; Defect; Development; Diabetes Mellitus; Dose; Endocrine Disruptors; Environment; Equilibrium; Exposure to; Faculty; Funding; Grant; Hormonal; Hormones; Human Milk; Hypospadias; In Vitro; Industrialization; Infant formula; Institution; Laboratories; Leydig Cell Tumor; Life; Life Cycle Stages; Long-Term Effects; Longitudinal Studies; Luteinizing Hormone; Male Genital Organs; Malignant neoplasm of testis; Mediating; Medical Students; Methods; Mitochondria; Molecular; Molecular Weight; Mouse Strains; Mus; North Carolina; Obesity; Oxidants; Oxidation-Reduction; Persons; Pharmaceutical Preparations; Plasticizers; Play; Polymers; Polyvinyl Chloride; Predisposition; Production; Progesterone; Proliferating; Proteins; Public Health Schools; Rattus; Reporting; Reproductive system; Research; Risk Factors; Role; Rural; Secondary to; Seminiferous tubule structure; Serum; Stem Cell Development; Steroids; Students; Surface; System; Testis; Testosterone; Time; Toxicant exposure; Training; Umbilical Cord Blood; Universities; aged; consumer product; dietary supplements; environmental stressor; flexibility; graduate student; in vivo; insight; interest; leydig interstitial cell; male; negative affect; neonatal exposure; phthalates; prenatal exposure; programs; response; self-renewal; stem; stem cell differentiation; stem cells; success; undergraduate student; underserved area

Project Summary By interfering with normal hormonal function and action, endocrine-disrupting chemicals (EDCs) such as the plasticizer di-(2-ethylhexyl) phthalate (DEHP) can have negative effects on the male reproductive system. In previous studies, Dr. Traore found that in utero exposure to DEHP can result in reduced serum testosterone levels in the adult. In studies conducted in vitro, he found that acute (short-term) exposure to mono-2-ethylhexyl phthalate (MEHP), a metabolite of DEHP, can inhibit testosterone production by isolated Leydig cells and progesterone production by MA-10 mouse tumor Leydig cells. Additionally, he found that MEHP can negatively affect mitochondrial function and cholesterol translocation in MA-10 cells. Until recently, it has not been feasible to conduct long-term studies using Leydig cells isolated from the testis because although these cells are able to produce high levels of testosterone, they do not do so for longer than 1-2 days. Therefore, it has not been feasible to use isolated Leydig cells to examine the direct, long-term effects of environmentally relevant concentrations of EDCs, or to critically evaluate mechanisms involved in phthalate actions. Dr. Zirkin’s lab at the Johns Hopkins Bloomberg School of Public Health (JHSPH) recently developed a method by which Leydig cells can be derived from the in vitro differentiation of stem cells associated with the seminiferous tubules, and that the derived Leydig cells are able to produce and sustain high levels of testosterone production for several months. The availability of these cells will be of great value for our proposed studies of the effects on testosterone production of acute, chronic, and repeat exposures of Leydig cells to environmentally relevant phthalate concentrations, and particularly so for addressing the molecular mechanisms involved in phthalate- induced reductions in testosterone. The studies that we propose in this R15 renewal application will: (1) increase our understanding of the mechanism by which environmental stressors mediate the inhibition of testosterone hormone production; and (2) provide new insights regarding multiple exposure of Leydig cells to phthalates, chemicals to which we are exposed throughout life; and 3) address whether early exposures predispose susceptibility to subsequent non-toxic dose exposure later in life. Campbell University has outstanding medical students, graduate students and undergraduates who will be involved in the proposed research. With funds from the REAP grant that the current application seeks to renew, Dr. Traore supported and trained many students in his lab at Campbell University, an institution located in a rural, underserved area of North Carolina. Renewal of the grant will allow Dr. Traore to continue and greatly extend ongoing research while increasing the interest in basic science and/or clinical research of the involved students. Moreover, his research success undoubtedly will influence attract other Campbell University faculty to seek funding to develop research programs in their labs.

项目摘要 通过干扰正常的激素功能和作用，内分泌干扰化学物质（EDC），例如 增塑剂Di-（2-乙基己基）邻苯二甲酸酯（DEHP）可能对男性复制系统产生负面影响。在 先前的研究，Traore博士发现，在子宫内暴露于DEHP中可能导致血清睾丸激素降低 成人的水平。在体外进行的研究中，他发现急性（短期）暴露于单2-乙基己基 邻苯二甲酸酯（MEHP）是DEHP的代谢产物，可以抑制分离的leydig细胞和 MA-10小鼠肿瘤细胞产生孕酮。此外，他发现MEHP可能负面 影响MA-10细胞中的线粒体功能和胆固醇易位。直到最近，它一直不可行 使用从睾丸分离的leydig细胞进行长期研究，因为尽管这些细胞能够 产生高水平的睾丸激素，它们不会超过1-2天。因此，这是不可行的 使用孤立的leydig细胞检查环境相关浓度的直接，长期影响 EDC，或严格评估参与邻苯二甲酸酯作用的机制。约翰·霍普金斯（Johns Hopkins）Zirkin博士的实验室 彭博公共卫生学院（JHSPH）最近开发了一种可以得出Leydig细胞的方法 从与半率管相关的干细胞的体外分化，并衍生的leydig 细胞能够生产和维持高水平的睾丸激素产生几个月。可用性 对于我们提出的对睾丸激素的影响的研究，这些细胞将具有很大的价值 Leydig细胞向环境相关的邻苯二甲酸酯的急性，慢性和重复暴露 浓度，尤其是解决邻苯二甲酸酯涉及的分子机制 诱导的睾丸激素降低。我们在此R15续签应用中提出的研究将： （1）提高我们对环境压力介导抑制的机制的理解 睾丸激素的生产； （2）提供有关Leydig细胞多次暴露于 邻苯二甲酸盐，我们一生都暴露的化学物质； 3）解决早期暴露是否 易感性的易感性对后续生命后来的无毒剂量暴露。坎贝尔大学有 杰出的医学生，研究生和本科生将参与该提案 研究。在当前申请寻求续订的收获赠款的资金中，Traore博士支持并 在坎贝尔大学实验室的许多学生培训了许多学生，该机构位于一个处于粗糙，服务不足的地区的机构 北卡罗来纳州。该赠款的续签将使Traore博士能够继续进行，并进行大量延长的持续研究 同时增加了对参与学生的基础科学和/或临床研究的兴趣。而且，他的 毫无疑问，研究成功将影响吸引其他坎贝尔大学教职员工来寻求资金发展 实验室中的研究计划。